final copy

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REMOVAL OF RED ACID DYE BY
SURFACTANT MODIFIED BAGASSE
Submitted in partial fulfillment of the
requirements for the degree of
Master of Technology
In
Biotechnology
by
Harshini E
(14MBT0060)
DEPARTMENT OF BIOTECHNOLOGY
SCHOOL OF BIO SCIENCE AND TECHNOLOGY
May 2016

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DECLARATION
I hereby declare that the thesis entitled “REMOVAL OF RED ACID DYE BY
SURFACTANT MODIFIED BAGASSE” submitted by me, for the award of the degree
of M.Tech in Biotechnology to VIT University, is a record of bonafide research work
carried out by me under the supervision of Dr. Sangeetha Subramaniam
I further declare that the work reported in this thesis has not been submitted and
will not be submitted, either in part or full, for the award of any other degree or diploma
in this institute or of any other institute or university.
Place: Vellore Signature of the Candidate
Date: 09-05-2016

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CERTIFICATE
This is to certify that the dissertation entitled “REMOVAL OF RED ACID
DYE BY SURFACTANT MODIFIED BAGASSE” submitted by Harshini E
(14MBT0060) to the VIT University, for the award of the degree of M.Tech., in
Biotechnology, is a record of bonafide work carried out by him under my supervision, as
per the VIT code of academics and research ethics.
The contents of this report have not been submitted and will not be submitted
either in part or in full, for the award of any degree or diploma in this institute or any
other institute or university. The thesis fulfills the requirements and regulations of the
University and in my opinion meets the necessary standards for submission.
Place: Vellore
Date: 09-05-2016
Signature of the Guide(s)

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Thesis Approval Form
This thesis, entitled
REMOVAL OF RED ACID DYE BY
SURFACTANT MODIFIED BAGASSE
and authored by “Harshini E”(14MBT0060), is hereby accepted and approved
External Examiner(s) Head of Department Dean SBST

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ACKNOWLEDGEMENT
I express my profound gratitude and heartfelt thanks to our Honorable Chancellor
Dr. G. Viswanathan for providing me an opportunity to carry out my research work in
the School of Bio Sciences and Technology, VIT University, Vellore.
I express my gratitude to Dr. C. Ramalingam, Dean School of Biosciences and
Technology, VIT University, for their support and guidance.
It is with deep sense of gratitude that I acknowledge the immense help and the
valuable guidance of my guide Dr. Sangeetha subramniam, SBST, VIT University,
Vellore. I thank her for giving me an opportunity to work under her and also for constant
encouragement, kindness and support.
My heart full thanks to all Teaching and Non-teaching members, Mr. Babu,
Mr. Karthikeyan, Mr. Velu, Mr. Suresh Kumar Lab technicians for their timely help
throughout my research.
Here I got a great chance to thank my dear friends Dr. Gopi kadiyala, Anitha J,
Esther Jo Vita, Pavan kumar C.
Last but not the least; I reward my sincere thanks to my parents for their constant
encouragement and support.
HARSHINI E

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ABSTRACT
Dyes are well known organic compounds used by various industries such as textile,
cosmetic, paper, leather, printing and rubber industries to color their products. Many of
these dyes are toxic, carcinogenic which cause skin and eye irritation. This study
investigates the potential use of sugarcane bagasse pretreated with surfactant extracted
from Reetha, Acacia and Planococcus for the removal of Acid red dye. The effects of
condition such as adsorbent dosage and pH were studied. It was observed that at 30
mins for pH 7(99.4%) and at 40ppm Bagasse treated with acacia shows maximum
removal of dye. According to this study acacia treated bagasse showed better
performance when compared to other two compounds.

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CONTENTS
S.no CHAPTER NAME
01 INTRODUCTION
02 AIM AND OBJECTIVE
03 REVIEW OF LITERATURE
04 MATERIALS AND METHODS
05 RESULTS AND DISCUSSION
06 CONCLUSION
07 REFERENCES

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LIST OF FIGURES
S.no NAME OF THE FIGURES
01 Bagasse
02 Reetha seeds
03 Acacia seeds
04 Map view of kumta coast
05 Planococcus
06 Extracted powder of Bagasse
07 Extracted powder of R10
08 Extracted powder of A10
09 Subculture Of P21
10 Microscopic view of P21
11 P21 inoculum
12 Parafilm M test
13 Surfactant extraction of R10,A10,P21
14 Bagasse treated with surfactants(P21,A10,R10)
15 Phenol test
16 Emulsified layer
17 Comparison of initial dye with BA10, BR10, BP21

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LIST OF TABLES
S.no NAME OF THE TABLES
01 Types of natural dyes
02 Types of synthetic dyes
03 Characteristics of Acid red dye
04 Dye separation and removal treatments
05 Raw and treated agricultural solid wastes
06 Bacterial and fungal strains
07 Composition of nutrient agar
08 Composition of nutrient broth
09 Composition of mineral salt medium
10 Composition of methylene blue agar plate
11 Biochemical test
12 Inference of Biochemical test
13 Drop collapse test
14 Parafilm M test
15 Surfactant extracted
16 E24% of P21, A10, R10

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17 Dye removal of BA10
18 Dye removal of BR10
19 Dye removal of BP21
20 LI of BA10
21 LI of BR10
22 LI of BP21
23 FI of BA10
24 FI of BR10
25 FI of BP21

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SYMBOLS AND NOTATIONS
RAD Red acid dye
MSM Mineral salt medium
MBAP Methylene blue agar plate
NA Nutrient agar
NB Nutrient broth
B Bagasse
A10 Acacia at 10g/100ml
R10 Reetha at 10g/100ml
P21 Planococcus isolated in lab no.212
E24 Emulsification activity
BA10 Bagasse treated with acacia
BR10 Bagasse treated with reetha
BP21 Bagasse treated with planococcus
MB Methylene blue
CTAB Cetyl trimethylammonium bromide
KCl Potassium chloride

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MgSO4 Magnesium sulphate
NH4Cl Ammonium chloride
CaCl2 Calcium chloride
CIES Chelated iron element solution
TES Trace element solution
FI Freundlich isotherm
LI Langmuir isotherm

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Textile industry is the fastest growing industry with the requirements of several
types of dyes for dyeing and printing of clothes. The effluent from these industries is the
major cause of environmental pollution, containing smaller amounts of dye[1]. are highly
detectable, causing environmental pollution and serious health hazards such as severe
headache, profuse sweating, mental confusion and similar health hazards[2].
Dye is a colored substance applied to fabrics to impart a permanent color,
which is not removable by washing or drying. W.H. Parkin was the first to use dye in
1856. As per the reports of Central Pollution Control Board (CPCB), commercially
produced dyes and its intermediates in India tends up to a million.
Based upon the chemical structure, dyes can be classified as,
a) Natural dyes
b) Synthetic dyes
1. a NATURAL DYES:
Natural dyes came in use since 1856, it was extracted from plants, animals, insects and
mineral sources. The most commonly used natural dye are tabulated below, [6]

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S.No NATURAL DYES SCIENTIFIC NAME
1 Berberry Berberis aristata
2 Cochineal Dacylopius coccus
3 Cutch Acacia catechu
4 Eucalyptus Eucalyptus globules
5 Flos Sophorae Sophora japonica
6 Henna Lawsonia inermis
7 Indigo Indigofera tinctoria
8 Indian Rhubarb Rheumemodi
9 Jackfruit Artocarpus heterophyllus
10 Onion Alium cepa
11 Teak Tectona grandis
12 Turmeric Curcuma longa
13 Weld Reseda luteola
Table no: 01 Types of natural dyes and its scientific names.
1. b SYNTHETIC DYES:
These dyes were discovered by W.H. Perkin. These dyes do not have complete degree
of fixation to fiber during dyeing process and finishing processes so it causes serious
hazards to the environment due to their synthetic origin and complex molecular structures
which decrease their ability to biodegrade. It is estimated that more than 100,000
commercial dyes are produced more than 7x105
tonnes per year [7].

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S.No NAME OF THE DYE BEST EXAMPLES APPLICATIONS
1 Acid dye Congo red, Congo orange,
Acid blue, Acid black, Acid
red, Acid yellow, Acid violet
Wool, Silk, Nylon
2 Basic dye Methylene blue, Crystal violet,
Basic red, Basic brown, Basic
blue, Anilline yellow, Brilliant
green
Polyester, Wool,
Silk, Nylon
3 Direct dye Martius yellow, Direct black,
Direct blue, Direct red, Direct
orange, Direct violet
Cotton, Wool, Flax,
Silk, Leather
4 Disperse dye Disperse blue, Disperse red,
Disperse orange, Disperse
yellow, Disperse brown
Polyamide fibers,
Wool, Polyamide
5 Reactive dye Reactive red, Reactive blue,
Reactive black, Reactive
yellow, Remazol
Cellulosic fibers,
Wool, Polyamide
6 Vat dye Indigo, Vat green, Vat blue Wool, Flax, Rayon,
Fibers
Table no: 02 Various types of dyes, its examples and uses[9].

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1.1 ACID RED DYE (AR):
1 C.I name 45100
2 CAS No 3520-42-1
3 Commercial name Acid Rhodamine B
4 Type of dye Acid dye
5 Chemical class Triphenyl methane
6 Description Dark Red Powder
7 Solubility (%) 80-85
8 Hue Bright bluish pink
9 Insolubility (%) 0.1 - 0.3
10 Ph 4.5 – 5.5
11 Strength – Reflectance 100 +/-2%
12 De 0.5 +/-0.2
13 Dye content (%) -----
14 Wavelength of maximum absorption 500 – 560 nm
15 Foaming Nil
16 Dusting property Dedusted
17 Moisture content 2 – 3
18 Fastness properties Light (2 – 3)
19 Washing 3
20 Perspiration 4
21 Dischargeability Poor

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22 Uses Dyeing of textiles (Wool, Xylon, Silk)
23 Non textiles Paper, leather
Table no: 03 Characteristics of Acid Red Dye[10]
1.1.1 PROPERTIES OF ACID RED DYE[20]:
 Anionic in nature.
 No affinity for cellulose.
 Suitable for wool, silk, polyamide and modified acrylics.
 Combine with the fiber by hydrogen bonds, vanderwaals forces and ionic
linkages.
 It is highly soluble in water.
1.1.2 ADVANTAGES:
 Found in bright colors.
 Widely used on silk.
 Used for protein fibers, nylon, spandex, special type acid dyeable acrylic
compounds.
1.1.3 DISADVANTAGES:
 Poor fastness in washing.
 It is banned in Europe as it was found to metabolise in the body back to
orthotolvidine.
 Many intermediates such as O-Tolvidine, Benzidine, etc., used in the
manufacture were found to be carcinogenic.

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CHAPTER 2
AIM AND OBJECTIVE

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2.1 AIM:
 To study and carry out the removal of Red acid dye(RAD) using three
surfactants treated with bagasse (BA10, BR10, BP21).
2.2 OBJECTIVE:
a) Extraction of surfactant from plant source and microbial source.
b) Treating of dye with agro-waste source such as Bagasse.
c) Treating of bagasse with surfactants like A10, P21, R10.
d) Comparison between three different surfactants treated with bagasse
such as BA10, BR10, BP21.

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CHAPTER 3
REVIEW OF LITERATURE

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Several methods for the removal of dye are discovered. Some of the available
techniques are:
a) Physico chemical treatment
b) Biological treatment
Physico chemical treatment includes Membrane filtration, Coagulation /
Flocculation, Precipitation, Flotation, Adsorption, Ion exchange, Ion pair extraction,
Ultrasonic mineralization, Electrolysis, Chlorination, Bleaching, Ozonization, Fenton
oxidation, Photocatalytic oxidation and Chemical reduction[18].
Biological treatment includes Bacterial and Fungal biosorption, Aerobic and
Anaerobic condition for combined and biodegradation process. Bacteria and Fungi are
proved as good adsorbents with biosorption capacity of 100mg dye/g biomass. Biological
treatments are cheap, naturally biodegradable that have a high binding capacity and act as
ion exchangers. Some of the biomaterials such as wheat, rice husks, bark, cotton waste,
cellulose, bacterial biomass, bagasse, etc., can be used as bioadsorbent. The main
advantage of bioadsorption is highly selective, efficiency, low cost and good removal
performance[19].

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S.NO TREATMENT
METHODOLOGY
TREATMENT
STAGE
ADVANTAGES
01 Precipitation,
Coagulation-
Flocculation
Pre/Main treatment a)short retention time
b)low capital costs
c)good removal efficiencies
02 Electro kinetic
coagulation
Pre/Main treatment Economically feasible
03 Fenton process Pre/Main treatment a)No volume alteration
b)Effective for soluble and
insoluble coloured compounds
04 Ozonation Main treatment a)Effective for removal of dye
b)No volume alteration
05 Silica gels Pre treatment Effective for basic dyes
06 Irradiation Post treatment Effective oxidation
07 Photochemical
process
Post treatment No sludge production
08 Electrochemical
oxidation
Pre treatment a)No additional chemicals are
required
b)Non dangerous
09 Ion exchange Main treatment Regeneration with low loss of
adsorbents

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10 Aerobic process Post treatment Partial or complete
decolorization for all types of
dyes
11 Anaerobic process Main treatment Resistant to complex colored
compounds
12 Single cell protein Post treatment a)Good removal
b)very effective
13 Membrane filtration Main treatment a)recovery and reuse of
chemicals and water
b)used for all types of dyes
14 Photo catalysis Post treatment Nontoxic and inexpensive
15 Sonication Pre treatment Simple to use
16 Enzymatic treatment Post treatment Effective for selected
compounds
17 Redox mediators Pre/Supportive
treatment
Easily available
Table no: 04, Various current and emerging dye separation and removal treatments along
with advantages.[10,12,14]

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S. No DYES ADSORBENTS ADSORPTION CAPACITY
(% of g/l)
1 Congo red Acid treated pine cone 45
Pine cone 20
Acid treated papaya seed 59
Organo-attapulgite 89
Papaya seed 71
Eucalyptus wood 67
Sugarcane bagasse 38
2 Methylene blue Date stones 43.5
Palm trees 39.5
Acid treated papaya seeds 60
Papaya seed 100
Pine cone 59.5
Pine tree leave 39.3
Palm kernel fiber 95.4
Garlic peel 79.3
Yellow passion fruit 44.7
Rice husk 40.6
3 Malachite green Neem bark 0.36
Mango bark 0.5
Peroxide treated rice husk 26

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4 Basic blue 3G Raw coffee residue 100
5 Tolvidine blue Coffee waste 72
6 Remazol blue Raw coffee residue 95
7 Reactive red 141 Palm shell 14
8 Reactive blue 21 Palm shell 24.7
9 Reactive black 5 Peanut hull 55.6
10 Acid black 26 Pine cone 62.9
11 Acid green 25 Pine cone 43.3
12 Acid blue 7 Pine cone 37.4
13 Basic red 46 Pine tree leaves 71.9
14 Crystal violet Palm kernel fiber 78.9
15 Indigo carmine Rice husk 65.9
16 Direct red Soy meal 60.3
17 Acid blue Soy meal 72.6
18 Acid red Soy meal 54.7
Table no: 05, Raw and treated agricultural solid wastes in dye removal[13,15,17]

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S. No DYES ADSORBENTS ADSORPTION
CAPACITY (% of
g/l)
1 Coracryl pink D. Flavida fungi 53
P. Chrysosporium fungi 100
D. Squalens fungi 100
2 Remozol blue T. Versicolor ATCC 98
P. Chrysosporium ATCC 95
Trametes species 89
3 Remozol red P. Chrysosporium 97
T. Versicolor ATCC 85
4 Coracryl violet P. Chrysosporium fungi 100
5 Congo red Bacteria consortium 90
6 Direct violet A.niger 92
7 Acid blue 62 C. polyzona 90
8 Blue BCC Bacterial consortium 74
9 Blue bezaktiv 150 Lyophilized bacterial consortium 62
10 Disperse red 1 T. Versicolor 50
11 Disperse blue 1 Myriconium sp. 43
12 Reactive red 4 S. Rugosoannulata 81
13 Coracryl black P. Sanguineus fungi 67
Table no: 06, Commonly used bacterial and fungal strains in biodegradation[15,16]

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3.1 FACTORS AFFECTING FOR DIFFERENT TECHNIQUES:
 Type of the dye.
 Composition of water.
 Chemicals, Energy and Materials required based on cost and dose.
 Environmental handling.
Exhausted research has been done from a very long period of time for the complete or
partial biodegradation of the dyes using pure or mixed culture of dye. According to our
own view, the present project is designed to explore the biosorption potential of
sugarcane bagasse treated with three surfactants (P21, R10, A10) for the removal of acid
red dye due to environmental problems. To our knowledge, this is the first study to
determine decolorization capacity of acid red dye by bagasse treated with three
surfactants.

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CHAPTER 4
MATERIALS AND METHODS

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General laboratory techniques were followed for preparation of media, inoculation and
maintenance of culture.
4.1 Cleaning of glassware’s:
All glass wares were immersed in cleaning solution for few hours. Then the glass
wares were washed thoroughly with tap water followed by detergent solution and
finally rinsed with distilled water. The cleaned glass wares were dried in hot air oven
and stored.
4.2 Sterilization:
Dried glass ware and media components were sterilized in an autoclave for 15min at
15 lb/sq. inch pressure.
4.3 Chemicals:
Analytical grade chemicals supplied by micro fine chemicals and Hi-media were used.
4.4 Plant collection:
The plants were collected from the nearby field with help of people with knowledge
about the plant.

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Figure 1: Bagasse(sugarcane waste)
Figure 2: Reetha (kunkudkai)

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Figure 3: Acacia (shikakai)
4.5 Organism collection:
Planococcus was collected from Protein engineering lab, VIT university and
subcultured for further use.
4.6 Preparation of the extract:
4.6.1 BAGASSE:
The waste of sugarcane after extracting juice is nothing but bagasse was collected and
washed with distilled water and dried under sun. Then it is cut into small pieces,
powdered with electric blender and stored for future use.

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Figure 4: Bagasse extract powder (B)
4.6.2 REETHA:
The seeds of Reetha was collected and washed with distilled water and dried under sun.
Then it is powdered with electric blender and stored for future use.
Figure 5: Reetha extract powder (R10)
4.6.3 ACACIA:
The seeds of Acacia was collected and washed with distilled water and dried under sun.
Then it is powdered with electric blender and stored for future use.

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Figure 6: Acacia extract powder (A10)
4.7 Media:
4.7.1 Composition of nutrient agar:
Ingredients gm Lit
Beef extract 1.0g
Yeast extract 2.0g
Peptone 5.0g
NaCl 5.0g
Agar 15.0g
Table 7: composition of nutrient agar

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4.7.2 Composition of nutrient broth:
Ingredients gm Lit
Beef extract 1.0g
Yeast extract 2.0g
Peptone 5.0g
NaCl 5.0g
Table 8: composition of nutrient broth
4.7.3 Composition of Mineral salt medium:
Ingredients gm Lit
NH4Cl 2.94 g
CaCl2 0.01 g
KCl 0.2 g
MgSo4 0.05 g
CIES 10 ml
TES 10 ml
Table 9: composition of MSM
4.7.4 Composition of MBAP:
Ingredients gm Lit
CTAB 5.0 g
MB 20 ml
Glucose 20.0 g
Table 10: composition of MBAP

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The media components were prepared for required quantity and kept for sterilization in
an autoclave for 15 min at 15 lb/Square inch pressure.
4.8 BIOCHEMICAL TEST [5, 10, 12]:
S.No NAME OF THE TEST INFERENCE
1 Phenol Presence of Glycolipids
2 Biuret Presence of Lipopeptides
3 Orcinol Presence of Rhamnolipids
4 Molisch’s Presence of Carbohydrates
5 Drop collapse Displacement of oil
6 Parafilm M Evaluation of droplets
7 Phosphate Presence of Phospholipids
8 MBP Presence of blue halozones
Table 11: Different test carried out to confirm the presence or absence of surfactant
4.9 EMULSIFICATION INDEX (E24):
Centrifugation at 10,000 rpm to separate biosurfactant from microbial culture which
yields a cell free broth. A mixture of 4:4 v/v of biosurfactant and crude oil is vortexed
at high speed for 20 minutes and stabilized for 24hrs so that emulsified layer is
formed. Emulsification activity is calculated by the formula,
E24(%)  (Height of emulsified layer  Total emulsified height)100

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4.10 EXTRACTION OF SURFACTANT FROM P21:
The culture inoculated in MSM along with crude oil and culture was centrifuged at
10,000rpm for 15 min, the supernatant obtained was added to 300 ml of acetone and is
stabilized for 24hrs for the formation of white brown sediment that is obtained after
centrifugation.
4.11 EXTRACTION OF SURFACTANT FROM R10:
10g of Reetha is diluted in 100ml of distilled water and placed on a magnetic stirrer
for 3hrs. Add 200ml of Methanol after centrifuging at 10000 rpm for 15mins and is
stabilized for 24hrs for the formation of dark brown precipitate that is obtained after
centrifugation[19].
4.12 EXTRACTION OF SURFACTANT FROM A10:
10g of Acacia is diluted in 100ml of distilled water and placed on a magnetic stirrer
for 3hrs. Add 200ml of Ethanol after centrifuging at 10000 rpm for 15mins and is
stabilized for 24hrs for the formation of dark brown precipitate that is obtained after
centrifugation[20].
4.13 BAGASSE TREATED REETHA:
2g of extracted R10 along with 2g bagasse is diluted in 100ml of water and kept in
orbital shaker for 24hrs. The medium is dried under hot air oven or sun shade after
24hrs and is used for further use[18].

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4.14 BAGASSE TREATED ACACIA:
2g of extracted A10 along with 2g bagasse is diluted in 100ml of water and kept in
4.15 BAGASSE TREATED PLANOCOCCUS:
2g of extracted P21 along with 2g bagasse is diluted in 100ml of water and kept in
4.16 EFFECT OF pH:
Effect of various pH 4,5,6,7,8 on dye decolorization of AR dye using B, BR10, BA10,
BP21 was studied for 120 min and was analyzed by taking absorbance at 510nm using
UV spectrophotometer[21].
4.17 EFFECT OF ADSORBANCE:
Effect of various adsorbent 20, 40, 60, 80, 100 (mg dye/l dist.H2O) on dye
decolorization of AR dye using B, BR10, BA10, BP21 at pH 7 was studied for 15
mins and was analyzed by taking absorbance at 510nm using UV spectrophotometer
and R2
value of Freundlich and Langmuir isotherm was calculated [22].
4.18 PERCENTAGE OF DYE REMOVAL:
Dye removal is used to calculate the removal of dye at particular time at particular pH
and concentration of the solution and is given by a formula[25],
% removal= [ (Control OD – Sample OD)/ (Control OD) ]  100

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CHAPTER 5
RESULTS AND DISCUSSION

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5.1 SUBCULTURE OF PLANOCOCCUS:
Figure 9: Colony of P21
Figure 10: Microscopic view using staining techniques shows that P21 is gram positive
Figure 11: Culture growth of P21 in Broth

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5.2 BIOCHEMICAL TEST:
S.No NAME OF THE TEST INFERENCE
1 Phenol Presence of Glycolipids
2 Biuret Absence of Lipopeptides
3 Orcinol Absence of Rhamnolipids
4 Molisch’s Absence of Carbohydrates
5 Phosphate Absence of Phospholipids
6 MBP Absence of blue halozones
Table 12: confirmation of presence or absence of surfactant
5.3 DROP COLLAPSE TEST:
S.NO NAME OF THE
MEDIUM
AT 0 min AT 1 min
1 P21 0.30 0.45
2 R10 0.40 0.60
3 A10 0.20 0.50
Table 13: Diameters evaluated at 0 min and 1 min for three surfactants
using drop collapse test

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5.4 PARAFILM M TEST:
Figure 7: Test done using parafilm M
At 0 min At 1 min
SDS (+ve) 0.3 cm 0.3 cm
Water(-ve) 0.4 cm 0.6 cm
Control 0.4 cm 0.4 cm
P21 0.3 cm 0.7 cm
R10 0.3 cm 0.8 cm
A10 0.2 cm 0.6 cm
Table 14: Diameters evaluated at 0 min and 1 min for three surfactants
using parafilm M test
5.5 EXTRACTION OF SURFACTANT:
Table 15: production of surfactant in g/l
Surfactant production (g/l)
P21 12.4
R10 20.5
A10 16.8

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Figure 8: surfactant extraction
5.6 BAGASSE TREATED WITH SURFACTANTS:
Figure 9: Bagasse treated with P21, A10 and R10
R10 A10 P21

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5.7 PHENOL TEST:
Figure 10: Test showing the presence of glycolipids
5.8 EMULSIFICATION ACTIVITY:
Figure 11: Upper layer is shown as Emulsified layer

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CONTROL P21 R10 A10
EMULSIFIED
LAYER
− 1.7 1.5 1.8
TOTAL
HEIGHT
3.9 3.9 3.9 3.9
E24% 0.0 43.5 38.4 46.1
Table 16: A10 has highest Emulsified layer than R10, P21
5.9 DYE REMOVAL:
Figure 12: Dye removal compared to control
Control (RAD)

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5.9.1 INITIAL OD OF 100ppm RAD WITH DIFFERENT pH:
pH OD AT 510nm
4 2.769
5 2.689
6 2.822
7 2.822
5.9.2 INITIAL OD OF DIFFERENT CONCENTRATIONS OF RAD AT
pH 7:
ppm OD AT 510nm
20 0.593
40 1.202
60 1.626
80 2.022
100 2.827

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5.9.3DYE REMOVAL OF BA10:
pH 30 min 60 min 90 min 120 min
4 91.09 92.50 92.50 95.9
5 94.5 94.5 95.5 96.0
6 95.5 95.8 95.9 96.2
7 97.1 97.2 98.5 98.9
Table 17: Dye removal of BA10 for different pH values.
Graph 1: % of dye removal vs. pH of BA10

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5.9.4 DYE REMOVAL OF BR10:
4 75.6 75.8 82.5 89.9
5 78.5 79.5 85.5 90
6 86.3 86.8 90.9 92.2
7 90.1 92.2 95.5 98.9
Table 18: Dye removal of BR10 for different pH values.
Graph 2: % of dye removal vs. pH of BR10

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5.9.5 DYE REMOVAL OF BP21:
4 75.6 75.8 82.5 89.9
5 78.5 79.5 85.5 90
6 86.3 86.8 90.9 92.2
7 90.1 92.2 95.5 98.9
Table 19: Dye removal of BP21 for different pH values.
Graph 3: % of dye removal vs. pH of BP21.

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5.9.6 Langmuir isotherm of BA10:
Ppm c Q 1/c 1/q
20 13.02 0.069 0.076 14.4
40 30.96 0.090 0.032 11.1
60 48.12 0.118 0.020 8.47
80 64.32 0.156 0.015 6.41
100 80.7 0.193 0.012 5.18
Table 20: LI of BA10 for different concentrations at 5 mins.
Graph 4: plot between 1/ c between 1/q of BA10

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5.9.7 Langmuir isotherm of BR10:
ppm ce qe 1/ce 1/qe
20 17.62 0.023 0.056 43.4
40 35.68 0.043 0.028 23.2
60 53.46 0.079 0.018 12.6
80 72.48 0.196 0.013 5.10
100 92.01 0.3 0.010 3.33
Table 21: LI of BR10 for different concentrations at 10 mins.
Graph 5: plot between 1/ c between 1/q of BR10

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5.9.8 Langmuir isotherm of BP21:
ppm ce qe 1/ce 1/qe
20 15.7 0.043 0.063 23.25
40 34.6 0.054 0.028 18.51
60 52.9 0.071 0.018 14.08
80 70.9 0.091 0.014 10.98
100 90.1 0.099 0.011 10.10
Table 22: LI of BP21 for different concentrations at 15 mins.
Graph 6: plot between 1/ c between 1/q of BP21

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5.9.7 Freundlich isotherm of BA10:
ce qe log ce log qe
13.02 0.069 1.11 -1.16
30.96 0.090 1.49 -1.04
48.12 0.118 1.68 -0.92
64.32 0.156 1.80 -0.80
80.7 0.193 1.90 -0.71
Table 23: FI of BA10 for different concentrations at 5 mins.
Graph 7: plot between log c between log q of BA10

54
5.9.8 Freundlich isotherm of BR10:
ce qe log ce log qe
17.62 0.023 1.24 -1.63
35.68 0.043 1.55 -1.36
53.46 0.079 1.72 -1.10
72.48 0.196 1.86 -0.70
92.01 0.3 1.96 -0.522
Table 24: FI of BR10 for different concentrations at 10 mins.
Graph 8: plot between log c between log q of BR10

55
5.9.9 Freundlich isotherm of BP21:
ce qe log ce log qe
15.7 0.043 1.19 -1.36
34.6 0.054 1.53 -1.26
52.9 0.071 1.72 -1.14
70.9 0.091 1.85 -1.04
90.1 0.099 1.95 -1.00
Table 25: FI of BP21 for different concentrations at 15 mins.
Graph 9: plot between log c between log q of BP21

56
DISCUSSION
 pH plays an important role in the adsorption process. As the pH and time
increases from 4 to 7, the percentage removal of ARD was increased in all three
surfactants.BA10 has more adsorption capacity when compared to BR10, BP21.
 Freundlich’s adsorption isotherm is better than Langmuir adsorption for all
three surfactants treated with bagasse.
 R2
=0.964 of BP21 has highest value in Freundlich’s isotherm when compared to
BA10(0.935), BR10(0.942).

58
The environmental problems created by the textile industries have received increased
attention for several decades because of contaminated effluents, which mainly arise from
dyeing processes. Due to toxicity of acid dyes, their removal from effluents has been an
urgent challenge. There is the search for efficient, ecofriendly and cost effective remedies
for waste water treatment. Attempts have been made through the process of biosorption ,
a green approach, to search novel biosorbents from plant sources, bacterial strain and
agro wastes as a tool to reduce percentage potential of contaminates in waste water. The
present project shows that if low cost adsorbents perform well in removing dye colour
they will be used widely in the industrial sector to improve profit and to minimize the
cost in efficiency in the industrial sector. These low cost biosorbents also offer a lot of
promising benefits for commercial purposes in the future. Various researcher are
working on removal of dyes using biosorption as a green approach , because of its low
cost ,no hazardous chemical formation .
Among three surfactants such as P21, A10, R10 treated with bagasse, BA10 shows
99.4% removal at pH 7 while BR10, BP21 shows 98.1 and 98.0 respectively at pH 7.
BA10 has fast degrading activity in short period of time when compared to BR10,
BP21.

60
1. Akhilesh, D., Neeraj, M, Neha, S, Abhina, S. and Shivendra, V. 2010. Isolation of
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63
APPENDIX
CHEMICALS
1. Methanol
2. Ethanol
3. EDTA
4. Sulphuric acid
5. Acetone
6. Chloroform
MEDIA
1. Nutrient agar
2. Nutrient broth
3. Methylene blue agar
DYE
1. Acid Red dye
OTHER MATERIALS
1. Bagasse
2. Reetha
3. Acacia

64
BACTERIAL STRAINS:
1. Planococcus
OTHER APPARATUS:
1. Weighing balance
2. Micro pipette
3. Petri dish
4. Centrifuge machine
5. Heater
6. Stirrer
7. Autoclave
8. Test tubes
9. Inoculating needle
10. Funnel
11. Whattman filter paper

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