Professional Documents
Culture Documents
A Project Report
Submitted at
Amity University, Noida
In partial fulfillment of the degree
Of
Bachelor of Technology
In
Biotechnology
1
Acknowledgement
There are people who simply by being what they are try to influence
you to do things which you could never thought of. Their spontaneous and
genuine
help as and when needed have helped me a lot, to gain profound knowledge
and experience in the analytical field.
2
ABBERIVATION
°C : Degree Centigrade
2Na2CO3 : Sodium bi Carbonate
AHR : Anaerobic Hybrid Reactor
AT : Aeration Tank
ATCC : American Type Culture Collection
BGA : Brilliant Green Agar
BOD : Biochemical Oxygen Demand
BPA : Baired Parker Agar
BSA : Bismuth Sulphide Agar
CA : Cetrimide Agar
CaCl2 : Calcium Chloride
CaCO3 : Calcium Carbonate
CB : Cetrimide Broth
CIP : Clean In Place
CO2 : Carbon dioxide
COD : Chemical Oxygen Demand
DDS : Dodecyl Sulphate
DM : De-Mineralized
DMF : Dual Media Filter
DO : Dissolved Oxygen
DOP : Dioctylphthalate
EMB : Eosin Methylene Blue
FC : Final Clarifier
FeCl3 : Ferric Chloride
FRP : Fibre Reinforced Plastic
GC : Gas Chromatography
GMP : Good Manufacturing Process
H2CO3 : Carbonic Acid
H2O : Water
H2S : Hydrogen Sulphide
H2SO4 : Sulphuric Acid
HCl : Hydrochloric Acid
K2HPO4 : Potassium Hydrogen Phosphate
KH2PO4 : Potassium dihydrogen Phosphate
KI : Potassium Iodide
KOH : Potassium Hydroxide
LAF : Laminar Air Flow
MgCl2 : Magnesium Chloride
MgCO3 : Magnesium Carbonate
MgSO4 : Magnesium Sulphate
MLSS : Mixed Liquor Suspended Solid
MnSO4 : Manganous Sulphate
MR-VP : Methyl Red – Voges Proskauer
MSA : Mannitol Salt Agar
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Na2HPO4 : Sodium Hydrogen Phosphate
NaEDTA : Sodium Ethylene Diamine Tetra
Acetic Acid
NaN3 : Sodium Azide
NaOH : Sodium Hydroxide
NH3 : Ammonia
NH4Cl : Ammonium Chloride
NMT : Not More Than
N-Tank : Neutralization Tank
OF : Oxidation / Fermentation
PC : Primary Clarifier
PDA : Potato Dextrose Agar
PIA : Pseudomonas Isolation Agar
PVC : Polyvenyl Chloride
QA : Quality Assurance
QC : Quality Control
RI : Refractive Index
RO : Reverse Osmosis
S.BLANK : Seeded Blank
SCDA : Soyabean Casein Digest Agar
SCOT : Support Coated Open Tubular
SHMP : Sodium Hexa Meta Phosphate
SS : Suspended Solid
STP : Sodium Tri Phosphate
SVI : Sludge Volume Index
TDS : Total Dissolve Solid
TPC : Total Plate Count
TSI : Triple Sugar Iron Agar
TSP : Tri Sodium Phosphate
UL : Upper Limit
UV : Ultra Violet
VJA : Vogel Johnson Agar
WCOT : Wall Coated Open Tubular
4
TABLE OF CONTENTS :
2) RAW MATERIAL
3) SANITATION
5) MANUFACTURING PROCESS
8) INVERSION
5
INTRODUCTION
Soft drinks constitute one of the largest beverage industries in the world today.
Tremendous advances have taken place in the process technology in the soft drink
industries in the past one or two decades.
The beverages are divided into two groups i.e carbonated soft drink like coke, thums
up, limca, fanta etc. & non-carbonated soft drink like maaza, minute maid.
The major ingredients of soft drinks are
Water
Sugar and/or sugar substitute
Carbon dioxide
Flavor emulsion and emulsifiers
Coloring agents
Acids and preservatives
Coca – Cola: An Insight
Our Roots
While much of the world has changed since 1886, the pure and simple magic of
one thing stays the same - COKE. The name and the product represent simple
moments of pleasure for consumers in nearly 200 COUNTRIES around the
globe, who reach for products of The Coca Cola Company hundreds of millions of
times every single day.
John Styth Pemberton first introduced The Refreshing Taste of Coke in Atlanta,
Georgia. It was May of 1886 when the pharmacist concocted a caramel-colored
syrup in a three-legged brass kettle in his backyard. He first ‘distributed’ the new
product by carrying Coin a jug down the street to Jacobs pharmacy. For five cents,
consumers could enjoy a glass of Coat the soda fountain. Whether by design or
accident, carbonated water was teamed with the new syrup, producing a drink that
was proclaimed
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‘Delicious and Refreshing’.
Dr. Pemberton’s partner and bookkeeper, Frank M.Robinson, suggested the name and
penner “Coke” in the unique flowing script that is famous worldwide today. Mr.
Robinson thought the “ two would look well in advertising”.
By 1891,
1891 Atlanta entrepreneur Asa G. Candler had acquired complete ownership
of the Coca-Cola Business. Within four years, his merchandising flair helped
expand consumption of Coca-Cola to every state and territory. In 1919, The Coca-
Cola Company was sold to a group of investors for $25 million. Robert W.
Woodruff became president of The Coca- Cola Company in 1923, and his more
than six decades of Leadership took the business to unrivaled heights of
commercial success, making Coca – Cola an institution the world over.
FIRST BOTTLED
COKE began as a fountain product, but candy merchant Joseph A. Bedenharn of
Mississippi was looking for a way to serve this refreshing beverage at picnics. He
began offering bottled Coke, using syrup shipped from Atlanta, during an
especially busy summer in 1894.
In 1899, large-scale bottling became possible when Asa Candler granted exclusive
bottling rights to Joseph B. Whitehead and Benjamin F. Thomas of Chattnooga.
The contract marked the beginning of The Company’s unique independent bottling
system that remains the foundation of Company Soft drink operations.
As the Company had many imitators, which consumers would be unable to
identify until they took a sip. The answer was to create a distinct bottle for Coke.
As a result, the genuine Coke bottle with the contour shape now known around the
world was developed in 1915 by the Root Glass Company.
7
• TRADEMARKS
The trademark “Coke” was registered with the US Patent & Trademark office in
1893, followed by “C” in 1945. The unique contour bottle, familiar to consumers
everywhere, was granted registration as a trademark by the US Patents &
Trademark office in 1977, an honor awarded to only a few other packages.
In 1982, The Coca – Cola Company introduced Diet Coke to US consumers,
marking the first extension of the Company’s most precious trademark to another
product. Later years saw the introduction of additional products bearing the name
of Coca-Cola, which now encompasses a powerful line of six cola products.
Today, the world’s favorite soft drink, Coke, is also the world’s best known and
most admired trademark, recognized by more than 90 PER CENT of the world’s
population.
8
Main building including Process areas , Packaging areas , 13096 m2
Warehouse , Stores & ETP
Utility 1755 m2
Caustic , HSD, Cooling towers, carbon dioxide , raw 3196 m2
water tanks
Admin. Block 918 m2
Empty Bottle storage yard 9600 m2
Car parking 490 m2
Truck Parking 1890 m2
Concrete roads 7500 m2
ETP 3000 m2
LPG store 120 m2
Driver amenities 64 m2
Contract Labour amenities 108 m2
Forklift repair area 100 m2
Security building 24.5 m2
Caustic storage area 100 m2
HSD storage area 324 m2
Carbon dioxide storage 1800 m2
Raw water storage area 720 m2
Switch yard 180 m2
9
Coca Cola, the name and the product represent simple moments of pleasure for
consumers in nearly 200 countries around the globe.
John Smith Pemberton first introduced the refreshing taste of coca cola in Atlanta,
Georgia. It was May of 1886 when the pharmacist concocted caramel coloured syrup
in a three-legged brass kettle in his backyard. He first distributed the new product by
carrying Coca-Cola in a jug down the street to Jacobs’s pharmacy. For five cents,
consumers could enjoy a glass of Cola-Cola at the soda fountain. Whether by design
or accident, carbonated water was teamed with the new syrup, producing a drink that
was proclaimed delicious and refreshing. Thus Coca Cola began as a fountain
product.
In 1899 large scale bottling became possible when Asa Candler granted exclusive
bottling rights to Joseph B. Whitehead and Benjamin F. Thomas of Chattanooga,
Tennessee. Today coca-cola products reach consumers and customers around the
world through a vast distribution network made up of local bottling companies. These
bottlers are located around the world, and most are independent businesses. Using
syrups, concentrates and beverage bases produced by the Coca-Cola Company. The
global bottling system packages and markets products, then distributes them to more
than 14 million retail outlets worldwide.
The trademark “Coca-cola” was registered with the U.S. Patent and Trademark office
in 1893, followed by “Coke” in 1945.
In 1982, the Coca-Cola Company introduced diet coke to U.S consumers, marking the
first extension of the Company’s most precious trademark to another product. Later
years saw the introduction of additional products bearing the Coca-Cola name, which
now encompasses a powerful line of cola products. Today, the world’s favourite soft
drink, Coca-Cola is one of the world’s best-known and most admired trademarks,
recognized by more than 90 percent of the world’s population.
10
• Taste or Odor Free of foreign taste or odor
a) PURITY
PROCEDURE :-
1. Fill the Zahm CO2 purity tester with water and observe for any air
bubbles
2. Attach the connecting tube to the sample valve of CO2 system
3. Open the stop cork between reservoir & body of tester.
4. Open the stop cork on the body of the tester before attaching the tubing
and allow water to flow out.
5. Quickly attach the tubing. Which has CO2 flowing through it, forcing
the water into reservoir
6. Allow the flow of gas to continue for about 30 sec.Close the stopcock
on the body of tester. Then close the stamping valve and reservoir
stopcock and detach tester from sampling tube.
7. Fill the reservoir with 30% w/v NaOH
8. Open the stopcock and allow the NaOH to flow into tester body.
Agitate the tester to be sure all CO2 is absorbed.
9. When the bubble remains constant in size close the reservoir stopcock.
10. Read % purity from the scale.
PROCEDURE:-
• Fruity
• Rotten egg, sewer,silage,sulfury
• Acetaladehyde
• Hydrogen sulfide
11
2. TESTS FOR SUGAR
a) TASTE
b) ODOR
1. Half fill a wide mouth screw capped bottle with dry sugar
2. Heat to 50 C
3. Smell and note the nature of any off odor
1. Smell the 50 brix solution at room temperature and note any off-odor
2. Add 0.2ml 75% w/v phosphoric acid to 50 ml of sugar solution in a
100ml glass beaker and mix
3. Cover the beaker with a watch glass and heat to 50C in a water bath or
incubator.
4. Smell the solution every 10 min for 30min and note the nature of any
off odor.
d) TURBIDITY
e) MICROBIOLOGICAL TESTING
1. Weigh 10g sugar in a sterile 250 ml flask and add sterile100 ml water
upto the mark. Cover with a foil and agitate to dissolve sugar.
2. Pour the solution into filter funnel.
3. Cover the funnel, apply vacuum.
4. Wash the flask twice with 250ml sterile water.
5. Transfer the membrane to a sterile petridish (M-TGE medium for total
count and M-green yeast and mold medium on schaufus – Pottinger
medium for yeast and mold count) and incubate at 35 C or 28 C
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6. for total count, count the colonies at 24hrs and at 72 hours. For yeast
and count , count colonies at 48 hrs and at 24 hrs interval thereafter
until 5 days have lapsed.
f) COLOR (ICUMSA)
Requirement:
• 3N HCL
• 0.02N EDTA solution (reagent A).
• 1N NaOH
• Hydroxy napthol blue.
13
• A conical flask.
• 500ml volumetric flask.
• A 50ml beaker.
• Distill water.
• 2 pipettes of 10ml.
Procedure
1. Weigh 1.5gm of lime powder in a beaker.
2. Add 10ml of distill water in above beaker.
3. Then add 30ml of 3N HCL.
4. Transfer it to conical flask for thorough mixing.
5. Take a 500ml volumetric flask.
6. Transfer contents of conical flask to volumetric flask, made up the volume by
distill water.
7. Put the cap of volumetric flask and mix thoroughly.
8. Pipette out 5ml of volumetric flask content into conical flask.
9. Add 10ml distill water, 1.5ml 1N NAOH and 0.30mg of Hydroxy napthol
blue.
10. Titrate it with EDTA solution.
11. End point – Blackish purple to blue.
Formula used:
Lime % = B.R × 3.705 × 100 × 0.02
1.5
Where,
B.R = Burette reading
Standard for lime % = 95
Requirements:-
• A conical flask.
• Distill water.
• 50ml beaker.
• Dropper.
• Measuring cylinder.
• Phenolphthalein indicator.
Procedure:-
1. Weigh 1.5g of caustic sample with the help of dropper in 50ml beaker.
2. Add 40ml distill water to the weighed sample and mix it thoroughly.
3. Transfer it to conical flask and add few drops of phenolphthalein indicator
4. Titrate it with 1N H2SO4 .
5. End point- pink to colorless
Formula used:-
Caustic % = B.R × 4
14
1.5
Where,
B.R = Burette reading
Standard for caustic % = 48
Requirements:-
• 4N H2SO4 .
• 3 N HCL.
• 1N NaOH.
• 0.1N KMnO4.
• 50ml beaker.
• 250ml volumetric flask.
• Funnel.
• Distill water.
• Conical flask.
• Ortho phosphoric acid.
• 2 pipettes of 10ml.
Procedure:-
1. Weigh 5gm sample in beaker.
2. Add distill water to weighed sample, mix it thoroughly & transfer it to
volumetric flask and make up the volume, again mix it.
3. Pipette out 50ml vol. flask content into conical flask, and then add 10ml
H2SO4 and 2ml ortho phosphoric acid.
4. Titrate it with 0.1N KMnO4.
5. End point – Colorless to light pink.
Formula used:-
FeSO4 % = 139 × B.R × 0.1
Wt. of sample taken
Where,
B.R = Burette reading
Standard for ferrous sulphate % = ≥ 97
PACKAGING MATERIAL
# Glass Bottles
# Crowns
15
# Pets
# Plastic caps
# Wrap around Labels
# Corrugated Cartons
# Shrink films
They are checked for weight dimensions, breaking strength compression
strength etc. The other details for expiry date, manufacturer date are printed on
the carton.
# RGB
(RETURNABLE GLASS BOTTLES)
1. Draw samples from min. 5 different crates / bulk packs with bottles of
different mould nos. for equitable sample representation
# CROWNS
Procedure for sampling of Crowns
3. Select 25 samples from each incoming consignment of 100 boxes (5 from each
of 5 boxes).
1. Critical defects.
• No liner.
• Deformed crowns
• Foreign crowns.(like Limca in ThumsUp)
• Foreign material.
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• Height out of specification(Go No Go)
• OD out of specification(Go No Go)
• Incorrect text(Std. color should be there)
2. Major defects.
• Rust
• Burrs.
• Die scratches.
• Excess liner
• Under fill liner
3. Minor defects.
• Litho damage.
• Litho off center.
• Lith off color.
• Scratched decoration.
• No decoration.
• Smudgy decoration.
• Liner adhesive poor.
# Corrugated Cartons
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• Maintain linkage between samples drawn and the consignment. This shall
enable resampling, resorting quarantine of the sampled lot, if required.
• Inspect the Carton as per the quality plan.
# Plastic closures
Procedure for sampling of plastic closure
SANITATION:
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The most important sanitation programme in the beverage plant deals with cleaning
and sanitizing of the surfaces that come in contact with syrup beverage, or ingredients
in their preparation.
Proper sanitation performed at the recommended frequency will minimize and most
likely completely eliminate the potential for bacteria, yeast and mold reproduction and
growth.
3 step CIP
5 step CIP
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a) Batch System
b) Continuous System
BATCH SYSTEM
Treated Water
Addition of sugar
Precoating
Cooling
Plate heat exchanger
Refrigerant-Glycol &cold
Water (by water-up to 35
Deg cent followed by glycol
22-40 Deg cent)
Simple raw syrup
Ready syrup
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Heating (PHE) to 85 Deg cent
Carbon Sugar
Contisolv
Filtration
Buffer tank
Bag Filter
Syrup Tank
Concentrate addition
Ready syrup
MANUFACTURING PROCESS
Depalletizing
21
Uncasing
Bottle Washer
Proportioner
Date Coding
Final Inspection
Casing
Palletizing
22
2) UNCASING: Removing of bottles from case. It is carried out by using uncaser
machine.
3) BOTTLE WASHING:Bottle washing is completed in following stages:
a) Pre-rinse:- Here empty bottles are rinsed by water used in first rinse.
b) First soak: - Bottles are soaked in surface active reagent solution like
caustic(2.8%) and Divo NEF ( 0.2-0.3%) . Here temperature is 60 deg cent and
pressure is 2-3 bars.
c) Second soak: - First soak is followed by second soak, here temperature is
maintained at 74 deg cent and pressure is at 2-3 bars.
d) Third soak: - Here again temperature is maintained as that of first soak.
This section wise soaking is carried to avoid thermal shocks to bottles .the
temperature difference between two soaks should not be more than 25 deg cent.
e) Rinse: - Here rinsing is carried out in four stages as first, second, third and final.
For rinsing, truated soft water is used. The flow of water is from final soak to dirst
soak. Water from first soak is used in pre rinse
Following are major issues and their reasons, during bottle washing.
a) Caustic carry over in bottles*
Chocked jets or disturbed jet alignment
Not proper water pressure.
Too high caustic dose.
Blowers off.
b) Excess breakage
Thermal shocks (high temperature difference between two sections of bottle washer)
Damaoed poskets.
c) M.B. Positive
Dirty bottles in feed
Caustic and additive percent is not up po limit.
Soak wise temperature difference.
23
Improper temperature and contect ti}e.
4) EBI (Empty Bottle Inspector): It is the new technique that is being used in coca
cola company.
It is used to detect 4 kinds of defects in the returnable glass bottles. In this cameras
and sensors are used to check the defects in the bottles.
Four kinds of defects are:
1) BASE :- it checks if the base is 3mm or not.
2) ISW ( Inner Side Wall) :- it checks if the inner side of the bottle wall is 3mm or
not.
3) IR :- it detects the level of water residue.
4) Radio Frequency :- it checks the level of caustic in bottle.
5) PROPORTIONER:
Machine used for mixing of syrup, carbon dioxide and water in proper
proportion is called as Proportioner
It contains following major parts:
Deairation
Proportioning
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Carbonating unit
Cooling unit
6) FILLING:
The operational principle is based on following functions
Positioning
Pressurizing
Filling
Closing & decompression
7) CROWNING:
Crowning applies the closure to the filled bottle.
The basic types of closures and closing equipment are
The function of the crowner is to mechanically apply and seal crowns to bottles. This
uses a crimping technique that applies pressure to the top and sides of the crown.
This pressure causes the crown to adapt to the neck of the bottle
The capper applies a pre-threaded plastic closure on the bottle, centers and pre
tightens the closure onto the bottle. The final stage seals to a pre-set dynamic torque.
When the pre set torque is reached, the clutch steps to prevent further tightening.
8) PACKAGE LABELLING:
Labellers are used to apply labels to returnable stock bottles and PET containers.
Labels can be used for special sales or promotions. Sleeve labels have the advantage
of more consistent operation with no glue application
Labels can be made of:
Plastic laminate
Paper
25
Combination of material
9) DATE CODING:
Date coding matter should include following information.
Date of production.
Time of production.
Batch number
Line identification.
The coder is installed on the production line and identifies the filled beverage package
as to establish
Production date
Regulatory requirements
Mandatory information
This information’s would allow plant to check back if there were any problems with
that production and to effectively manage the age of inventory in trade.
.10) FINAL INSPECTION:
The beverage filled bottles are again passed through a manual inspection station
where the beverage is scrutinised for appearance, clarity, presence of any particulate
matter, and half filled bottles. Rejected bottles are removed before packaging into
cases
11) CASING:
Mechanically by using caser machine casing is carried out.
12) WAREHOUSING:
The filled bottles are arranged in cases and through a belt conveyor system are taken
to the shipping or warehouse area where they are stored till they are marketed.
• It is the total arrangement made with the object of ensuring that beverage
products are of the quality required for their intended use.
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a) Beverages are designed and developed in a way that accounts of the
requirements of GMP and associates codes such as those of good laboratory
practice GLP and good clinical practice (GCP).
f) Beverages are not sold or supplied before the authorized person have certified
that each production batch has been produced and controlled in according with
the requirements of the label claim and any other regulations relevant to the
production, control and release of pharmaceutical products.
g) Satisfactory arrangements exist to ensure, as for as possible, that the
pharmaceutical products are stored by the manufacturer, distributed and
subsequently handled so that quality is maintained through out their shelf life.
h) There is a procedure for self – inspection and/or quality audit that regularly
appraises the effectiveness and applicability of the QA system.
MICROBIOLOGY LAB
To distinguish the food of acceptable quality from food of unacceptable quality
required the application of what are known as microbiological criteria. Three different
types of microbiological criteria have been identified.
27
1) A microbiological standard is a criteria specified in a law or regulation. It is a
legal requirement that foods must meet and is enforceable by the appropriate
regulatory agency.
• AUTOCLAVE
28
An Autoclave
29
• INCUBATOR
An Incubator
An incubator comprises a transparent chamber and the equipment that regulates its
temperature, humidity, and ventilation. For years, the principle uses for the controlled
environment provided by incubators included hatching poultry eggs and caring for
premature or sick infants, but a new and important application has recently emerged,
namely, the cultivation and manipulation of microorganisms for medical treatment
and research. It is used for providing favorable temperature conditions for the growth
of culture organisms. Generally the temperature of incubator is operated at 37°C for
the growth of microorganisms
Laboratory incubators were first utilized during the twentieth century, when doctors
realized that they could be could be used to identify pathogens in patients' bodily
fluids and thus diagnose their disorders more accurately. After a sample has been
obtained, it is transferred to a Petri dish, flask, or some other sterile container and
placed in a rack inside the incubator. To promote pathogenic growth, the air inside the
chamber is humidified and heated to body temperature (98.6 degrees Fahrenheit or 37
degrees Celsius). In addition, these incubators provide the amount of atmospheric
carbon dioxide or nitrogen necessary for the cell's growth. As this carefully
conditioned air circulates around it, the microorganism multiplies, enabling easier and
more certain identification.
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• CENTRIFUGE
A centrifuge is a piece of equipment, generally driven by a motor, that puts an object
in rotation around a fixed axis, applying force perpendicular to the axis. The
centrifuge works using the sedimentation principle, where the centripetal acceleration
is used to separate substances of greater and lesser density. There are many different
kinds of centrifuges, including those for very specialised purposes
It is used to separate the suspended matters as pallets/button/residue from the liquid as
supernatant.
• MICROSCOPE
A Microscope
A microscope is an instrument for viewing objects that are too small to be seen by the
naked or unaided eye. The science of investigating small objects using such an
instrument is called microscopy. The term microscopic means minute or very small,
not visible with the eye unless aided by a microscope. The microscopes used in
schools and homes trace their history back almost 400 years
31
• pH METER
A simple pH meter with its probe immersed in a mildly alkaline solution. The
two knobs are used to calibrate the instrument
32
• HOT AIR OVEN
It is used for dry heat sterilization. Glassware’s, Petri plates and pipettes are
packed in stainless steel containers and kept at 180°C for 2 hrs.
• BOD INCUBATOR
A BOD Incubator
33
• CYCLOMIXER
It is used for dissolving sample without the destruction of the organism for
which the cost is to be carried out. Sample + dilution is placed in the
recommended bags provided the total volume should be with in recommended
capacity of the machine (80 – 400 ml).
• WEIGHING BALANCE
34
VALIDATION OF EQUIPMENTS
35
• MICROBIOLOGICAL TESTING:
Different types of media used in microbiology lab to test microbiological count in
various samples.
Water being the essential component of beverage industry is tested against microbes
using various media.
Standard formula:
Yeast extract : 5.00 gms/ltr
Dextrose : 20.00 gms/ltr
Chloromphenicol : 0.10 gms/ltr
Agar :14.90 gm/ltr
36
2. Violet Red Bile Agar:
Standard Formula:
Peptic digest of animal tissues : 7.00 gms/ltr
Yeast extract : 3.00 gms/ltr
Lactose : 10.00 gms/ltr
Bile salts mixture : 1.50 gms/ltr
Nacl : 5.00 gms/ltr
Neutral Red : 0.03 gms/ltr
Agar : 15:00 gms/ltr
pH (at 25 c ) 7.4 + 0.2
Directions : Suspend 41.53 gms of salt in 1000 ml distilled water . Heat to boiling to
dissolve the medium completely. Cool to 45 c and immediately our into sterile petri
plates containing the innoculum. If Desired , medium can be sterilized by autoclaving
at 15lbs pressure at 121 c temperature for 15 mins.
Use : For selective Isolation , detection and enumeration of Coli – aerogens bacteria
in water, milk and other dairy food products.
37
3.EMB Agar :
Standard formula :
Peptic digest of animal tissues : 10.00 gms/ltr
Dipotassium phosphate: 2.00 gms/ltr
Lactose : 5.00 gms/ltr
Sucrose : 5.00 gms/ltr
Eosin –Y : 0.40 gm/ltr
Methylene Blue : 0.005 gms/ltr
Agar : 13.50 gms/ltr
Directions: Suspend 36 gms in 1000 ml distilled water. Heat to boiling to dissolve the
medium completely. Dispense and Sterilse by autoclaving at 15lbs pressure at 121 c
temperature for 15 mins.
Use : Used for differential isolation of Gram-ve enteric bacilli from clinical and non-
clinical specimen.
38
4. CC2:
Standard Formula:
Yeast extract: 9 gms/ltr
Cerelose : 50 gms/ltr
Bio Peptone : 10 gms/ltr
Magnesium Sulphate : 2.10 gms/ltr
Potassium sulphate: 2.00 gms/ltr
Diastase : 0.05 gms/ltr
Thiamine : 0.026 gms/ltr
Bromo Cresol Green Agar : 15.00 gms/ltr
pH (at 25 c ) 4.6+0.2
Directions: Suspend 8.82 gms in 1000 ml distilled water . Mix thoroughly. Heat to
boiling to dissolve the medium completely. Dispense and Sterilse by autoclaving at
12-15lbs pressure at( 118- 121 c) temperature for 15 mins.
Use : For counting yeast and molds in samples by membrane filter method.
39
Media used for checking and controlling microbial count in MAAZA.
Standard Formula:
Casien enzymatic Hydrolysate: 10 gms/ltr
Yeast Extract : 3 gms/ltr
Dextrose : 4 gms/ltr
Dipotassium Phosphate : 2.50 gms/ltr
Orange Serum Agar ( solid from 200 ml): 17 gms/ltr
pH (at 25 c) 5.5+0.2
Directions: Suspend 45-5 gms in 1000ml Distilled water . Heat to boiling to dissolve
the medium completely. Dispense and Sterilse by autoclaving at 12-15lbs pressure
at( 118- 121 c) temperature for 15 mins. Avoid overheating . Mix well and pour into
Sterile Petri plates.
Use: For Cultivation and enumeration of micro- organisms associated with spoilage
of citrus products. Cultivation of Lacto bacilli and other aciduric organism and
pathogenic fungi.
40
INVERSION:
INVERTED BRIX: This is the brix which is found after breaking of sucrose in two
parts and molecular weight increases due to water molecule addition in the presence
of acid.
C12H22O11 + H2O C6H12O6 + C6H12O6
(SUCROSE) (GLUCOSE) (FRUCTOSE)
(342) (180) (180)
(A) (B) {360}
41
WATER TREATMENT PLANT
INTRODUCTION
The bottling plant receives its water supply from 3 bore wells .This water is first
treated and then used for beverage preparation.
42
STANDARS OF WATER AT COCA COLA COMPANY
43
WATER TREATMENT PLANT TESTING
1) Physical Parameter:
TDS
Odour
Taste
Turbidity
Appearance
2 )Chemical Parameters:
Calcium hardness
Sulfate
Iron
Total Hardness
Total and Partial Alkanity
Chloride
Free/Total Chlorine
Bore wells
44
TREATED WATER
Coagulation Tank
.
Lime, bleaching pd., FeSO4
Dechlorination
Lead ACF
Lag ACF
5 micron filter
UV chamber
3 micron filter
1 micron filter
Treated water
45
USE OF TREATED WATER
Syrup making, beverage preparation and CIP.
Filler for cleaning and flushing.
Water coolers.
Back washing of PSF and ACF of treated water
SOFTENING STREAM
Raw water
P.S.F.
A.C.F.
Soft water
46
KINLEY WATER MANUFACTURE
Water from bore well
Coagulation
PSF
Storage tank
Dechlorination ACF
UV chamber
Reverse osmosis
Storage tank
Micron filter
Ozonation
Labeling
Date coding
Packaging
Palletizing
47
VARIOUS WATER TREATMENTS
48
Sanitation – Done by 50 ppm chlorine solution.
5) Softener
Scope – To reduce the hardness of water.
Medium Used ® Sodium resins
Resin0Quantity –"1500 ltr. ( Ex: Indion 225)
Working(Priociple – Na+ will be exclcnged with the hardness causing
elemunts.MRegeneration
Fepends on the hardness of output wa|er (generally every 4 days)
Done0by 2<% brine (NaCl) solution.
Sanitation!:% As per fresuency & depends on micro count
6) Polishing Filtration
Scope
Removes granular activated ccrbon$particles and sand(particles
Flakes of scale or rust
Media Used – Micron filters ( 5, 3,1 & 0.2)
Reverse Osmosis
7) Ultra Violet Purification System
Scope – Destroys or inactivates the DNA thus preventing micro organisms from
reproducing.Media – Ultra violet lamp radiation of 2537 Angstrom units.
49
8) Reverse osmosis
Reverse osmosis (RO) is most versatile technology available to bottling and canning
plants today
It reduces alkalinity and total dissolved solids by more than 90 %
Reduces inorganic such as Na, Cl, SO4
Reduces large organic molecules and organisms at efficiency more than 99%
The usual molecular weight range is below 300 Dalton
Operating pressure range is from 200 – 450 psi.
Advantages:
Highly effective against a wide spectrum of contaminants.
Easy to operate.
Cost effective when designed properly
Minimum space requirement
Can handle changes in water supply and level of impurities.
Disadvantages:
Polyamide membranes must be protected against effects of chlorine.
Cellulose acetate membranes are biodegradable.
Efficiency of unit affected by the temperature of raw water.
Higher the organic content of incoming water, less effective the unit.
The effluent waste water from system pose disposal problems.
50
9) Decaustisizer:
Decaustisizer
Bottle Washer
PHE
(Plate Heat Exchanger)
Chlorination
Settling Tank
Overflow Tank
PSF
ACF
Chlorine dozing
Degaser
51
Rinsing water bottle washer
1. Chloride test:-
(removes turbidity)
Where,
R – Reading
52
2. Sulphate test:-
100ml sample
2 drops of P. indicator
53
End point – Blue color
3. Total hardness:-
Procedure:-
Mix thoroughly
(Pink color appears)
54
4. Alkalinity:-
Procedure:-
55
5. Chlorine test:-
Procedure:-
Kept it in colorimeter
Observe color
56
6. Calcium Hardness:-
Procedure:-
7. Magnesium hardness:-
Procedure:-
Same as Calcium hardness
MgH = TH – CaH
Where,
MgH – Magnesium hardness
TH - Total hardness
CaH – Calcium hardness
57
8. Turbidity:-
• Collect the sample in a clean dry glass beaker, transfer the sample
to the sample cell quickly
• Rinse the sample cell with the water to be tested
• Cap the cell and dry the outside surface of the cell with a tissue
paper
• Pour the sample into the sample cell
• Examine the water sample in the cell before placing it in the
instrument. If bubbles have formed on the inside wall of the cell;
gently tapping on the cell wall or mildly agitate the cell to release
the bubbles
• Gently invert sample once to agitate any particulate that may
have settled
• Place the cell in the turbidity meter with the direction mark on the
cell forward
• Lower the light cover and the turbidity will be displayed
• Record the turbidity
9. P – test (alkalinity):-
58
EFFLUENT TREATMENT PLANT:
INTRODUCTION –
Treatment plants remove impurities contained in wastewater so that the treated
wastewater can be safely returned to the environment. This same stabilization process
occurs in nature to break down wastewater into its most basic components of carbon
dioxide and water. Common methods of treatment include physical, biological and
chemical treatment steps to stabilize the wastewater.
The effluent treatment facility is installed for biological treatment of the effluent
emanating. The effluent bears large amounts of organic matter. The direct discharge
of the effluent into the water bodies causes depletion, of DO of the water. Hence, in
order to meet the recommended standards of quality of the effluent, it is necessary to
treat the effluent before it is finally disposed off. This treatment facility provides for
removal of major pollutants from the effluent.
There are three reasons why most companies consider on – site treatment of
wastewater: -
a) To avoid prosecution
b) To remove restriction on the output of the factory
c) To save money
d) To protect public health in the service area
e) To protect the water quality in the waterway which receives the treated
effluent from the processes.
f) To protect the environment which receives any residuals from the
treatment processes.
Industries carry out cost/benefit studies to show how to achieve the greatest benefit
from the investment in effluent treatment. They design plants for the treatment of
59
industrial effluents tailored to the requirements of the site and the industrial process,
and they can arrange a complete service through to installation and commissioning of
the effluent plant.
Wastewater from industrial processes can be difficult to treat. The cost of disposing
of the effluent to the public sewer is determined by the volume, the polluting load, the
suspended solids in the flow and the treatability of the effluent.
It may not be possible to treat the effluent with municipal sewage, or it may be cost-
effective to treat the effluent on site. The plant may be designed to reduce the strength
of the effluent to a level suitable for discharge to the sewer, or to a standard suitable
for discharge to the environment, or to optimise the balance between on-site costs and
disposal charges.
Industrial wastewaters are typically much stronger than domestic sewage, and require
a different approach if they are to be treated economically.
Sludge disposal is becoming one of the greatest problems both for the industrial
wastewater treatment plants. The available routes for disposal are reducing rapidly,
and costs are escalating.
Modern aerobic treatment plants produce far less sludge with a smaller footprint on
the site. Anaerobic plants produce minimal sludge with a by-product of methane,
which can be used in the upstream processes for heating or power generation.
60
TREATMENT PROCESS –
PROCESS CONCEPT –
The raw effluent, bears large amount of suspended solids and oxygen consuming
organic matter. The conceptual approach of the treatment includes the removal of
suspended particles, dissolved organic matters and handling of sludge for disposal.
The heart of this treatment scheme is the aerobic biological reactor, which are
designed on the basis of activated sludge process. The activated sludge treatment
process basically involves the stabilization of organic matter by the action of various
microorganisms as depicted in the following equation.
In this biological process, a part of the newly synthesized sludge undergoes oxidation
called, Endogenous respiration.
The preformed biological flocks (MLSS) come in contact with the incoming waste in
the aeration tank under highly aerobic environment, and oxidize the organic matter to
more stable materials. The efficiency of the system mainly depends upon the
concentration of active microorganism present to perform the assimilation of organic
matter. The activated sludge, in general, consists bacteria and protozoan, rotifers etc.
in the presence of DO. The desirably environmental condition like sufficient DO,
substrate and nutrients are required for cell growth and energy for various metabolic
functions. It is essential that the biological flock should readily separate from the
treated wastewater in the final clarifier.
61
The oxygen supply is required for the following: -
1. Oxidation of organic matter (substrate removal)
3. Nitrification
Nitrosomonas
2 NH₄ + 3O₂ 2 NO₂ + 2H₂O + 4 H⁺
Nitrobacter
2 NO₂‾ + O₂ 2 NO₃‾
Excess or deficient quantity of food (incoming BOD) adversely affects the physical
quality of biological sludge. The activated sludge system is designed on the basis of a
particular food to microorganism ratio. This ratio is in practice indicated by the
quantity of BOD in influent per unit quantity of mixed liquor suspended solids per
unit time. This may be expressed as kg, BOD/kg, MLSS/day. The volatile suspended
solid, which repression is between 60 – 70% of MLSS is used as a measure of active
cells in the system. The optimal pH for an active biological aeration system is
between 6.5 – 9.0.
The surplus biological sludge (and the sludge from the secondary clarifier) needs
further dewatering, which is achieved in sludge drying beds. The final effluent is
suitable for discharging into the inland surface water.
62
PROCESS UNITS –
The units are designed for maximum of efficiency within certain flow range and
effluent characteristics. Close control and co–ordination of the operation of different
units are required within limits of design.Efficient plant operation is possible only
when the operator is fully conversant with the equipments and function of each unit.
This effluent treatment facility consists of the following units: -
1) Storage tank
2) Equalization tank
3) Neutralization tank
4) Primary clarifier
5) Anaerobic Hybrid Reactor
6) Aeration tanks – 1 & 2
7) Final clarifier – 1
8) Final clarifier – 2
9) Sludge drying beds
1) STORAGE TANK -
OBJECT –
The function of storage tank is that it collects and store the raw effluent from
different part of the factory.
PROCESS –
The raw effluent is collected from the different part of the factory and stored. The
storage tank is of 40 feet in height. The capacity of the tank is two lack liters. Now
from the storage tank the raw effluent is passed to the equalization tank with the help
of pump. The pH of the raw effluent in the storage tank is 5.5 – 6.5, which generally
come out from the factory.
63
2) EQUALISATION TANK -
OBJECT –
The function of equalization tank is to equalize the raw effluent emanating from
different processing units.
PROCESS –
3) NEUTRALIZATION TANK -
OBJECT –
The function of the neutralization tank is to neutralize the raw effluent, which is
generally acidic in nature.
PROCESS –
The raw effluent, which is usually acidic (pH-5.5 to 6.5) in nature is neutralized by
adding the saturated solution of NaOH, So, the final pH of the neutralization tank is
adjusted to pH- 8.0 to 9.0. Then the raw effluent after has been treated in
neutralization tank is allowed to passed in the primary clarifier through gravity.
4) PRIMARY CLARIFIER –
OBJECT –
The function of PC is to remove suspended heavy particles from the raw effluent.
PROCESS –
In this tank, the heavy particles along with the sludge, which the bacteria have
degraded settles down at the bottom of the tank and the water flows on top of it. A
rotator is fixed in the middle of the tank, so that the heavy particle along with the
64
sludge which has been settle down does not block the outlet of the PC. In this tank
mostly the inactive heavy particles along with little amount of sludge is thrown out in
the Sludge drying beds. The pH of the PC is maintained to 7.0 to 8.0.
OBJECT –
PROCESS –
The effluent after treated in PC is passed to the AHR through gravity. The design of
the AHR is in a way that at the bottom of this tank anaerobic bacteria’s beds are
made. The effluent which comes from PC react with the anaerobic bacteria and the
break up of organic compounds takes place with the production of Methane gas
which can be seen in the form of bubbles on the upper layer of the water in the tank.
The pH of the AHR is maintained to 7.0-7.5 because the anaerobic bacteria are stable
in this pH. If there is much fluctuation in the pH of this tank the anaerobic bacteria
can die.
65
6) AERATION TANKS 1 & 2 -
OBJECT –
This unit is provided for aerobic biological treatment of the effluent for the reduction
of organic matter in the effluent.
PROCESS –
The effluent from the AHR is received in the aeration tank stage-1 by pumping and is
aerated by the help of “OXYRATOR” mechanical surface aerators in the presence of
previously developed biological sludge (Mixed Liquor Suspended Solids i.e. MLSS).
The food / microorganism ratio is maintained at about 0.6 and 0.137 in the first and
second stage aeration tanks respectively which correspond to about 3500 mg / ml.
OPERATION –
The start up of the activated sludge process can be accomplished by using seed
sludge available from night soil develop a suitable microorganism population
expressed as MLSS.
The following method is recommended for the initial development of MLSS in the
aeration tank: -
The use of seed sludge (Night soil) provides the reliable means of start up. Seed
sludge may be added in the aeration tank to provide approx. 500mg/ltr. MLSS. The
tank is to be filled up with fresh water prior to the addition of seed sludge. The seed
sludge is to be aerated by running both the aerators and be continued for at least 24
hrs. in order to make the sludge into aerobic. With the seed sludge aerated, raw
effluent into the aeration tank is to be introduced at approx. 25% of the design flow.
If possible, aeration must be continued by all aerators and feeding of effluent
increased in daily increments of 25%. If there is no indication of the process
deterioration. This enables the treatment process to produce a quality effluent as the
MLSS concentration is increasing. During this operation also be added the requisite
quantity of nutrients in aeration tank.
66
Required nutrients viz. N and P are added with aeration tanks by pumping a solution
of Urea and DAHP. The aerators also help to keep the biological solids in suspension.
The mixed liquor from the aeration tanks is subjected to gravitational settling in the
hopper bottom secondary clarifier.
7) FINAL CLARIFIER 1 -
OBJECT –
The function of final clarifier-1 is to separate biological solids from the mixed liquor
first stage aeration tank.
PROCESS –
The mixed liquor from the first stage aeration tank is received in the clarifier by
gravity. The clarifier is hopper bottom type. The sedimentation of sludge is
withdrawn by pumps and is recirculated back into the aeration tank stage-1 for
maintaining the MLSS. Provision is given to transfer the sludge into the stage-2
aeration tank through the necessary connections given on the delivery line of the
sludge recirculation pump.
OPERATION –
The clarifier is filled up with effluent by gravity. The biological solids get settled by
gravity at bottom. Keep the suctions valves corresponding to each hopper portion of
clarifier open. Recirculate the settled sludge by operating pump back into the aeration
tank continuously. If the MLSS exceed the required level, or sludge needs to be
wasted, divert the sludge into aerobic.
8) FINAL CLARIFIER –2 -
OBJECT –
The function of final clarifier-2 is to separate the biological sludge from the mixed
liquor from the aeration tanks before the final effluent is disposed off.
PROCESS –
The mixed liquor from the aeration tank is received in the clarifier by gravity. Final
clarifier-2 is a circular sedimentation tank with the central chute inlet peripheral
overflow laundar. The sedimentation of sludge takes place by gravity setting. The
settled sludge is collected to a central circular channel around the inlet chute by a
rotating scarper. Scraper is driven by a central drive head. The settled sludge is
67
pumped back into the aeration tank. The clarified effluent from the annular laundar is
disposed off through the V- Notch.
68
Allow the sludge to flow to the drying beds. Once the sludge thickness comes to about
300 mm charging of sludge is to be stop and the bed is isolated to dry up by natural
evaporation. This takes about 10 days.
After drying and dewatering, the sludge cakes are removed manually and are
disposed off.
SPECIFICATION OF ETP -
69
6. MLSS AT-1 NMT 2000 ppm
TESTING -
GENERAL –
Full spectrum of water and wastewater testing can be performed to evaluate the
specific characteristics of water, wastewater or treated effluent. The ability to
determine what is happening within a plant, including evaluations of plant
performance, can only be done when proper sampling, storage and transportation
techniques have been followed
It is imperative to analyze regularly the operational parameters and maintain a
systematic record as a ready reckonar. Sampling and testing should be done as per the
methods prescribed in:
1) Standard methods for the examination of water and wastewater. (APHA,
AWWA, WCPC 1975)
2) Manual for the examination of water, sewage and industrial waste.
(ICMR)
3) Methods of sampling and test for sewage and industrial effluent.
SAMPLING POINTS –
70
METHODS OF ANALYSIS -
pH -
The pH of water refers to its hydrogen ion activity and is expressed as the logarithm
of the reciprocal of the hydrogen ion activity in moles per litre at a given temperature.
The practical pH scale extends from 0 (Very acidic), to 14 (Very alkaline), with 7
corresponding to exact neutrality at 25°C. Whereas alkalinity and acidity are
measures of the total resistance to pH change or buffering capacity of a sample, pH
represents the free hydrogen ion activity.
PRINCIPLE –
Although the hydrogen electrode is recognized as the primary standard, the glass
electrode is less subject to interferences and is used in combination with a calomel
reference electrode.
The glass reference electrode pair produces a change of 59.1 mg/pH unit at 25°C.
APPARATUS –
1) Electronic pH meter with temperature compensation arrangement.
2) Glass electrode; are available for measurement over the entire pH range
with minimum sodium ion-error types for high pH- high sodium samples.
3) Reference electrode; Use calomel, silver-silver chloride or other constant
potential electrode.
PROCEDURE –
Firstly, calibrate the pH meter with the buffer solution of pH -7.0 and then the pH -
4.0. After calibrating it the electrode is washed with DM water and finally the pH is
taken of the sample. After doing the work again the electrode is washed with DM
water and then the electrode is dipped in DM water.
One-Day Analysis: -
Raw – 6.70
PC – 7.28
71
AHR – 7.20
FC – 8.6
N-TANK – 8.58
SUSPENDED SOLID –
Estimation of suspended solid plays a important role for the process evaluation. These
solids are mostly of organic species and contributes pollutants load to the treatment
system. SS is analysed once in a week.
PRINCIPLE –
This test is based on the evaporation of the residues obtained after filtering a known
volume of sample, to dryness under standard conditions and weighing the residue
after drying.
APPARATUS –
Gooch Crucibles - 50 ml. Capacity
Measuring cylinder - 100 ml. Capacity
Vacuum pump
Dry heat Oven
SAMPLE –
72
CALCULATION –
Volume of sample
One-Day Analysis: -
73
MIXED LIQUOR SUSPENDED SOLID (MLSS) –
PRINCIPLE –
The tests are based on the evaporation of the mixed liquor sample to dryness under
standard conditions and weighing the residue after drying. MLSS is the weight of
residue, of the known filtered mixed liquor, on evaporation at 103 to 105°C.
APPARATUS –
SAMPLE –
AT 1 - 50 ml.
AT 2 - 50 ml.
74
PROCEDURE –
CALCULATION –
One-Day Analysis: -
75
SLUDGE VOLUME INDEX –
SVI VALUE –
APPARATUS –
76
Measuring cylinder - 1000 ml.
SAMPLE –
AT –1 and AT –2 - 1000 ml.
PROCEDURE –
CALCULATION –
One-Day Analysis: -
97 X 1000
SVI = = 74.61
1300
77
CHEMICAL OXYGEN DEMAND –
The COD determination provides a measure of oxygen equivalent of that portion of
he organic matter in a sample that is susceptible to oxidation by a strong chemical
oxidant. In the absence of a catalyst however, this method fails to include some
organic compounds (such as acetic acid), which are biologically available to the
stream organisms, while including some biologic compounds, which are not part of
the immediate biochemical load on the oxygen assets of the receiving water.
The use of exactly the same technique each time is important because only a
part of the organic matter is included; the proportion depending upon the chemical
oxidant used the structure of the organic compounds and the manipulative procedure.
The dichromate reflux method has been selected for the COD determination
because it has advantages over other oxidants in oxidizability, applicability to a wide
variety of samples and ease of manipulation.
The basis for the COD test is that nearly all organic compounds can be fully oxidized
to carbon dioxide with a strong oxidizing agent under acidic conditions. The amount
of oxygen required to oxidize an organic compound to carbon dioxide, ammonia, and
water is given
This expression does not include the oxygen demand caused by the oxidation of
ammonia into nitrate. The process of ammonia being converted into nitrate is referred
to as nitrification. The following is the correct equation for the oxidation of ammonia
into nitrate.
The second equation should be applied after the first one to include oxidation due to
nitrification if the oxygen demand from nitrification must be known. Dichromate does
not oxidize ammonia into nitrate, so this nitrification can be safely ignored in the
standard chemical oxygen demand test.
78
PRINCIPLE –
A boiling mixture of chromic and sulphuric acids destroys most types of organic
matter. A sample is refluxed with known amounts of potassium dichromate and
sulphuric acid, and the excess dichromate is titrated with ferrous ammonium sulphate.
The amount of oxidizable
organic matter, measured, as oxygen equivalent is proportional to the potassium
dichromate consumed. COD is analyzed daily.
APPARATUS –
a) Reflux apparatus consisting of a flat bottom 250 to 500 ml. capacity flask
with ground glass joint and condenser with 24/40 joint.
b) Hot plate
c) Titrator
d) Reflux flasks
e) Simple flask
f) Pipette
REAGENTS –
79
3) Ferron Indicator – Dissolve 1.735 gm of 1, 10 phenonthroline
dihydrate together with 695 mg ferrous sulphate crystalline, in DM water and
dilute to 100 ml.
4) Silver sulphate
5) Mercuric sulphate
SAMPLES –
Raw - 1 ml.
PC - 2 ml.
AHR - 5 ml.
FC - 10 ml.
PROCEDURE –
a) Firstly, take known quantity of samples in reflux flasks.
b) Then, 20 ml. DM water in Blank and others make the volume 20 ml. with
DM water.
c) After that add 400-mg. mercuric chloride, 10 mg. silver sulphate, 10 ml.
potassium
dichromate and finally add 30 ml. concentrated sulphuric acid.
d) Keep it for 2 hrs. for reflux on reflux apparatus.
e) Then cool it for 30 minutes.
f) After cooling add 100 ml. DM water.
g) Then, titrate with 0.1 N ferrous ammonium sulphate using ferrion
indicator. Take as the end point the orange color change to blue, then green
and lastly to reddish brown, even through the blue- green may reappear within
minutes.
CALCULATION –
(a-b) N X 8000
COD in mg. /liter =
ml. Sample
Where:
One-Day Analysis: -
80
PC = 22.8 2 X 0.1008 X 8000 / 2 = 806.4mg/ltr.
AHR = 22.7 2.1 X 0.1008 X 8000 / 5 = 338.68mg/ltr.
FC = 22.6 0.9 X 0.1008 X 8000 / 10 = 96.76 mg/ltr.
81
BIOCHEMICAL OXYGEN DEMAND –
The Royal Commission on River Pollution which was established in 1865 and the
formation of the Royal Commission on Sewage Disposal in 1898 led to the selection
in 1908 of BOD5 as the definitive test for organic pollution of rivers. Five days was
chosen as an appropriate test period because this is supposedly the longest time that
river water takes to travel from source to estuary in the UK.
PRINCIPLE –
Biochemical Oxygen Demand is defined as the amount of O₂ required by
microorganism while stabilizing biologically decomposable organic matters in a
waste under aerobic conditions. The BOD test is widely used to determine: -
2) The degree of pollution in lakes and streams at any time and their self-
purification capacity.
3) Efficiency of sewage treatment plant.
82
BOD Level (in ppm) Water Quality
1-2 Very Good-not much organic waste present
3-5 Moderately clean
6-9 Somewhat polluted
10+ Very polluted
APPARATUS –
a. Incubation bottles: Use glass bottles having 300 mL capacity. Clean bottles with a
detergent, rinse thoroughly, and drain before use. As a precaution against drawing air
into the dilution bottle during incubation, use a water seal. Obtain satisfactory water
seals by inverting bottles in a water bath or by adding water to the flared mouth of
special BOD bottles.
REAGENTS –
83
1) Phosphate buffer – Dissolve 8.5 gm KH₂PO₄, 21.75 gm K₂HPO₄, 33.4
gm Na₂HPO₄. 7H₂O and 1.7 gm NH₄Cl in DM water and dilute to 1000 ml and
adjust pH to 7.2
2) Magnesium sulphate – Dissolve 22.5 gm MgSO₄ .7H₂O and dilute to
1000 ml.
3) Calcium Chloride – Dissolve 27.5 gm of Anhydrous Calcium Chloride
and dilute to 1000 ml.
4) Ferric Chloride – Dissolve 0.25 gm FeCl₃ .6H₂O and dilute to 1000 ml.
SAMPLES –
CALCULATION –
One-Day Analysis: -
84
Raw = 6.9 5.1 (1.5 – 0.2) X 300 / 1 = 480 mg/ltr.
AHR = 6.7 4.9 (1.8 – 0.2) X 300 / 2 = 240 mg/ltr.
FC = 6.8 5.6 (1.2 – 0.2) X 300 / 30 = 10 mg/ltr.
DISSOLVED OXYGEN -
PRINCIPLE –
Dissolved oxygen analysis measures the amount of gaseous oxygen (O2) dissolved in
an aqueous solution. DO is one of the most important indicators of the quality of
water for aquatic life. O₂ dissolves freely in water as a result of photosynthesis,
community, respiration, diffusion at the air water interface, and wind driven mixing.
Temperature, pressure and salinity determine the amount of DO water can hold, or its
saturation level. DO concentration below 3.0 mg/l are generally consider harmful to
aquatic life, but requirements vary according to species, temperature, life stage,
activities and concentration of dissolved substances in the water. When performing
the dissolved oxygen test, only grab samples should be used, and the analysis should
be performed immediately. Therefore, this is a field test that should be performed on
site.
ENVIRONMENTAL IMPACT:
Total dissolved gas concentrations in water should not exceed 110 percent.
Concentrations above this level can be harmful to aquatic life. Fish in waters
containing excessive dissolved gases may suffer from "gas bubble disease"; however,
this is a very rare occurrence. The bubbles block the flow of blood through blood
vessels causing death. External bubbles can also occur and be seen on fins, on skin
and on other tissue. Aquatic invertebrates are also affected by gas bubble disease but
at levels higher than those lethal to fish.
APPARATUS –
a) BOD bottles
b) Measuring cylinder
c) Titrator
d) Iodine flask
e) Pipette
REAGENTS –
85
1) Manganous Sulphate – Dissolve 36.4 gm of MnSO₄ .H₂O and
dilute to 100 ml. filter if necessary. This solution should not give color with
starch when added to an acidified solution of KI.
2) Alkali Iodide-Azide – Dissolve 500 gm of NaOH and 150 gm of
KI and dilute to 1000 ml with DM water. Add 10 gm of sodium azide (NaN₃)
dissolved in 40 ml of DM water. This solution should not give color with
starch solution when diluted and acidified.
3) Starch Indicator – Prepare paste of 0.5 gm starch powder in DM
water. Pour the solution in 100 ml boiling water, allow to boil fpr few minutes.
cool and then use.
4) Sodium thiosulphate 0.025 N – Dissolve 6.25 gm of sodium
thiosulphate in boiled and cooled DM wsater, dilute to 1000 ml preserve by
adding 5 ml chloroform. Standardize before each titration.
METHOD –
CALCULATION –
86
REFERENCES:
10. Internet
87