Project Presentation on Optimization of Copper Removal Using Biomass

Project Presentation
on
By
ASHA.S.HEGDE (1DS07BT010) DIVYA.G.KAMBI (1DS07BT014) KAVYA.S (1DS07BT021) VIII Semester, B.E Biotechnology
DAYANANDA SAGAR COLLEGE OF ENGINEERING

SM. Hills, KS Layout, Bengaluru -560 078
Under the guidance of Nagamani. S. K, (Guide)

Lecturer, Department of Biotechnology Dayananda Sagar College of Engineering SM. Hills, KS Layout, Bengaluru -560 078
Shwetha. N, (Co-Guide)
Lecturer, Department of Biotechnology Dayananda Sagar College of Engineering SM. Hills, KS Layout, Bengaluru -560 078
OBJECTIVES
To identify the effects of
Initial copper ion concentration on the percentage removal of

copper pH on the percentage removal of copper
Temperature on the percentage removal of copper

Biomass loading on the percentage removal of copper To statistically optimize these variables for maximum removal of copper efficiently
14-06-2011
Dept of Biotechnology
INTRODUCTION
Bioadsorption : Physiochemical process that occurs naturally in
certain biomass which allows it to passively concentrate and bind contaminants onto its cellular structure.
Biomass :
Economical alternative for removing toxic heavy metals
Practiced in environmental cleanup Heavy metal characteristics

Nondestructive Toxicity Bioaccumulation Subsequent bio magnifications Non-Biodegradable in nature
Dept of Biotechnology 3
14-06-2011
Sources of heavy metals

Mining operation Electroplating operations Metal processing Coal fire powered Generation Nuclear industry Other industrial operations
Metal plating industry

Discharges huge amounts of copper Causes acute and chronic disorders in humans Major concerns because of their toxicity and threat to human life and environment
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Heavy metal pollution

Dangerous to health and environment (e.g. mercury, cadmium, lead, chromium). Causes corrosion (e.g. zinc, lead). Carcinogenic in nature, affecting Central nervous system (manganese, mercury, lead, arsenic). Kidneys (mercury, lead, cadmium, copper). Liver (mercury, lead, cadmium, copper). Skin, bones, or teeth (nickel, cadmium, copper, chromium).
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Department of Biotechnology
Effects of Heavy metals

Lead :
Acute or chronic damage to the nervous system
Cadmium :
Long term exposure - Renal dysfunction High exposure - Obstructive lung disease
Copper :
High doses - Anemia, liver and kidney damage, stomach and intestinal irritation
Chromium :
Low-level exposure Skin irritation (ulceration) Long-term exposure Kidney dysfunction
Selenium and Mercury :
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Damage to brain, circulatory tissue and central nervous system

Department of Biotechnology 6
COPPER
Symbol : Cu Atomic Number : 29 Atomic Mass : 63.546 amu Melting Point : 1083.0C (1356.15 K, 1981.4 F) Boiling Point : 2567.0C (2840.15 K, 4652.6 F)
Continued.
Copper : Essential compound for plants ,animals and humans in small quantities Toxicity : Excess bioaccumulation
Cupric ion : Main ionic form of copper, highly toxic

Cupric ion in water : Bound with inorganic and organic compounds, Reduces cupric ion concentrations
Physical and chemical properties
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Conventional Techniques
Precipitation Ion-Exchange Method Electrochemical Cells Reverse Osmosis
Biological Methods
Bioremediation Phytoremediation
Advantages of Biosorbents:
Cost Effectiveness High quality of treated effluent Minimization of chemical and or biological sludge No additional nutrient requirement Regeneration of bioadsorbent and Possibility of metal recovery
Solid Phase Extraction (Adsorption) is an attractive technique based on the sorbent that retains analytes.
Adsorbents used:
Peat Marine algae Clays Maize cob Bagasse Palm fruit bunch Lalang leaf Saraca Indica leaf and Nile rose plant
Metal adsorption reactions:

1. Bulk partitioning relationships Freundlich Isotherms Langmuir Isotherms 2. Surface complexation model (SCM) Effects of changing pH, solution composition and ionic strength.
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Adsorption Isotherm
Freundlich isotherm
Adsorption over a wider range of metal ion concentration.
max
La
m ng
r ui
Fr
dl e un
ich
The adsorption relationship is expressed as:
=K cN
Where, N is a fitting parameter.
log =log K +N log c
log[c]
R max is the max sorbate uptake under the given conditions
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Langmuir Isotherm
Adsorbent contains a finite number of reactive site on the surface. The metal ion sorption reaction can be expressed by a site-specific equilibrium reaction: [A- ]+[M+] [ AM] An equilibrium constant can be determined from the law of mass action:
K
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A M

13
AM
Kinetics of Bioadsorption

Metal sorption by microbial biomass often involves two distinct stages. The first stage the passive adsorption to the cell surface ( a rapid process). The second stage is slower and commonly involves diffusioncontrolled Accumulation.
Mechanism of Bioadsorption
Most organic functional groups are Amphoteric. R-AH R-A- + H+ The ionization of functional groups in the cell wall provides an electrical charge at the cells surface.
Dissociation of functional group on the cell surface

Organic acid: R-COOH R-COO- + H+
Hydroxyl group: R-OH R-O- + H+ Phosphate group: R-PO4H2 RPO4H- + H+
MATERIALS AND METHODS

Neocuproine method
Principle:
Copper in neutral or slightly acidic solution . Reacts with neocuproine to form a complex in which 2 moles of neocuproine are bound by 1 mole of Cu+ ion. The complex extracted by chloroform-methanol (CHCl3-CH3OH) mixture, to give a yellow solution at 457 nm. The color follows Beers law up to a concentration of 0.2 mg Cu/25 ml solvent Full color development -pH of aqueous solution is between 3 and 9. The color is stable in CHCl3-CH3OH for several days.
Stock solution of copper

To 200.0 mg polished electrolytic copper wire or foil in a 250-ml conical flask, add 10 ml water and 5 ml conc. Nitric acid. After the reaction has slowed, warmed gently to complete dissolution of the copper. Cooled & added 50 ml water, transferred to a 1litre Volumetric flask, and diluted to the mark with double distilled water; 1 ml = 200 g Cu.
Copper reacting with nitric acid

Source:www.uncp.edu/home/mcclurem/ptable/copper/
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Standard copper solution:

Dilute 50.00 ml stock copper solution to 500 ml with water;1.00 ml = 20.0 Cu
Sulfuric acid
Hydroxylamine-hydrochloride solution:
Dissolve 50 g NH2OHHCl in 450 ml water.
Sodium citrate solution:

150 g in 400 ml.
Ammonium hydroxide Neocuproine reagent Chloroform

Calibration curve
Blank : 50 ml water was pipetted into a 125-ml separatory funnel for use as a reagent. 1.00ppm to 200ppm standard copper solution were prepared and taken into a series of separating funnels.
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Standardization values and graph

Chromium Concentration (ppm) 1 2 3 4 5 6 7 8 9 10 20 30 40 50 60 70 80 90 100 150 5/5/2013 200 Optical Density 0.003 0.005 0.007 0.014 0.017 0.022 0.031 0.031 0.038 0.037 0.073 0.112 0.149 0.192 0.235 0.282 0.331 0.381 0.44 0.504 0.581
Calibration
Series2
0.7
0.6
0.5
Optical Density
0.4
0.3
0.2
0.1
0 1 2 3 4 5 6 7 8 9 10 11 12 Concentration(ppm)
r value = 0.9271
20
Figures showing the samples prepared for calibration
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Apparatus:
Colorimetric equipment: One of the following is required: 1) Spectrophotometer: for use at 457 nm, providing a light path of 1 cm or longer. 2) Filter photometer: providing a light path of 1 cm or longer and equipped with a narrow-band violet filter having maximum transmittance in the range 450 to 460 nm.
Separatory funnels
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Bioadsorbents
Tamarind fruit shell (Tamarindus indica)
Carrot peels (Daucus carota)
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Preparation of Adsorbent
1. Carrot (Daucus carota) Peels Collected from Carrot pods. Powdered and sieved to get uniform size (60-80) mesh particle size. Washed thoroughly with distilled water, acid, base and dried in the oven for 2 hrs at 60C. 2. Tamarind (Tamarindus indica) fruit shells Collected from tamarind fruit pods. Powdered and sieved to get uniform size (60-80) mesh particle size. Washed thoroughly with distilled water, acid, base and dried in the oven for 2 hrs at 60C.
Batch studies: Following parameters were varied in a

batch study
Initial metal ion concentration ( 1ppm 100ppm)

pH (1 7) Temperature ( 30C - 50C )
Biomass loading ( 1g/l 20g/l)
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Batch Studies
Bioadsorption studies:
Carried out using a certain amount of carrot and tamarind biomass in copper nitrate solution.
The operating conditions such as pH, adsorbent amount, contact time and metal concentration were investigated. For each of the investigation, the mixture was shaken in a rotary shaker at 180 rpm followed by filtration using Whatman filter paper. The filtrate containing the residual concentration of Cu (II) was determined spectrophotometrically at 457 nm using the method mentioned above.
Cu Ion Determination
A yellow colored complex was developed in the reaction between Cu(II) and 2,9-dimethyl-1,10-phenanthroline hemihydrate (Neocuprione) in Acidic condition. The percentage of Cu removal due to Bioadsorption was calculated as, % Cu removed = [(Co Ci) / Co] x 100% Where, Co - Initial conc. of Cu solution (mg/l) Ci - Equilibrium conc. of Cu solution (mg/l)
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Chemicals and Equipments

The chemicals and equipments used in the experimental work along with their specifications are given in the tables:
Chemicals
Hydroxylamine hydrochloride Sodium Citrate LR S.d.fine-Chem.Ltd.
Grade
LR
Suppliers
S.d.fine-Chem.Ltd
Neocuprione
Chloroform Methanol Isopropyl alcohol Copper Foil
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LR
LR LR LR
S.d.fine-Chem.Ltd.
S.d.fine-Chem.Ltd S.d.fine-Chem.Ltd S.d.fine-Chem.Ltd
Specification of materials Department of Biotechnology
28
Equipments Used
Name of the Equipment Name of the supplier/company
Weighing Balance
AY220 - SHIMADZU
Shaker
Hot air oven UV double beam spectrophotometer pH meter
POOJA LAB EQUIPMENTS, MUMBAI

FATHA INSTRUMENT WORKS UV VISIBLE 1700 - SHIMADZU MICROPRO
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Sieving machine
UV spectrophotometer
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Shaking Incubator
RESULTS AND DISCUSSION

The bioadsorption of metals using carrot peels (Daucus carota) and tamarind (Tamarindus indica) fruit shell powder in a batch process depends on the test variables considered. The effect of contact time on percentage removal of copper is studied by conducting the separate experiments designed using software minitab 14 and 15.
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Software Minitab version 15

Response surface methodology(RSM) Empirical statistical technique employed for multiple regression analysis of quantitative data obtained from statistically designed experiment. In this study full factorial central composite design using response surface methodology was employed
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pH 7 4 4 7 4 4
Initial conc (ppm) 1.0 50.5 50.5 100 48.5 50.5
Biomass loading (g/lt) Temperature ( C) 1.0 10.5 1.0 1.0 10.5 10.5 50 40 50 50 40 40
1
4 7 1 4 7 1 7 4 7 7 4
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100
50.5 100 1.0 49.5 100 1.0 100 50.5 50.5 1.0 50.5
20.0
10.5 20.2 1.0 10.5 20.0 20 1.0 10.5 10.5 1.0 10.5
30
40 30 30 40 50 30 30 40 4 30 40
34
Continued
pH 1 4 7 1 1 4 4 2 7 4 1 1 4 Initial conc (ppm) 100 50.5 1.0 1.0 100 50.5 50.5 50.5 1.0 50.5 100 1.0 50.5 Biomass loading (g/lt) Temperature ( C) 20.0 10.5 20.0 1.0 1.0 10.5 8.5 10.5 20.0 20.0 1.0 20.0 10.5 50 50 50 50 50 40 40 40 30 40 30 50 40
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Range of parameters designed using minitab software
Temperature- 30C, 40C, 50C.

Initial concentration- 1ppm, 48.5ppm, 50.5ppm, 100ppm Biomass loading- 1g/l, 10.5g/l, 20g/l Ph- 1, 2, 4, 7
Effect of Initial concentration- Tamarind

95.5 95 94.5 94
Ph-4 Temperature- 30C
Biomass loading- 10.5g/l
%Recovery
Series1 93.5 93 92.5 92 91.5 91 0 20 40 60 80 100 120 Initial concentration

Series 1- distilled water wash Series 2- Acid wash Series 3- Base wash
Series2 Series3
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Effect of initial concentration- Carrot

Ph-1 Temperature- 30C Biomass loading- 10.5g/l
Series 1- distilled water wash Series 2- Acid wash Series 3- Base wash
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Effect of Temperature
Effect of Temperature(Carrot Distilled water wash)
Ph- 1
95.5
Initial conc- 50.5ppm

95 94.5
Biomass loading -20g/l
%Recovery
94 Series1 93.5 93 92.5 92 0 10 20 30 40 50 60 Temperature('C)
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Effect of Temperature(Carrot HCl wash)

96.65 96.6 96.55 96.5
Ph- 1 Initial conc- 50.5ppm Biomass loading -20g/l
%Recovery
96.45 96.4 96.35 96.3 96.25 96.2 96.15 0 10 20 30 40 50 60 Temperature('C) Series1
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Effect of Temperature(Carrot NaOH wash)

95.1
Ph- 1
95 94.9
Initial conc- 50.5ppm Biomass loading -20g/l
%Recovery
94.8 Series1 94.7 94.6 94.5 94.4 0 10 20 30 40 50 60 Temperature('C)
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Effect of Temperature- Tamarind

95.5 95 94.5 94
Ph- 4 Initial conc- 50.5ppm Biomass loading -20g/l

Tamarind Distilled water wash Tamarind HCl wash Tamarind NaOH wash
%Recovery
93.5 93 92.5 92 91.5 91 90.5 0 20 40 60 Temperature('C)
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Effect of pH
Effect of pH
95.5 95 94.5
Temperature- 40C Initial conc- 100ppm Biomass loading- 10.5g/l
%RECOVERY
94 93.5 93 92.5 92 91.5 0 2 4 pH 6 8 Series1
Effect of pH on copper adsorption using Tamarind Distilled water

Effect of pH(Tamarind HCl wash)

95.5 95 94.5
%Recovery
94 Series1 93.5 93 92.5 92 0 1 2 3 4 pH 5 6 7 8
Effect of pH on copper adsorption using Tamarind Distilled water

Effect of pH(Tamarind NaOH wash)

96 94 92
%Recovery
90 88 86 84 82 80 0 1 2 3 4 pH 5 6 7 8 Series1
Temperature- 40C
Initial conc- 100ppm

Effect of pH on copper adsorption using Tamarind NaOH wash
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Effect of pH(Carrot Distilled water wash)

97 96.5 96
Temperature- 40C
Initial conc- 100ppm

%Recovery
95.5 Series1 95 94.5 94 93.5 0 1 2 3 4 pH 5 6 7 8
Effect of pH on copper adsorption using Carrot Distilled water wash

Effect of pH(Carrot acid wash)

95.5 95 94.5 94
%Recovery
93.5 Series1 93 92.5 92 91.5 91 0 1 2 3 4 pH 5 6 7 8
Effect of pH on copper adsorption using Carrot acid wash

Effect of pH(Carrot NaOH wash)

95.5 95 94.5 94
%Recovery
93.5 93 92.5 92 91.5 91 90.5 90 0 1 2 3 4 pH 5 6 7 8 Series1
Effect of pH on copper adsorption using Carrot NaOH wash

Effect of Biomass
Ph- 4 Temperature- 40C Initial conc 48.5ppm
Effect of Biomass loading on copper adsorption using Tamarind
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Ph- 4 Temperature- 40C Initial conc 48.5ppm
Effect of biomass loading on copper removal using carrot residues

Effect of Time
Effect of time on percentage removal of copper using tamarind distilled water wash sample at 30C
Effect of time on percentage removal of copper using tamarind acid wash sample at 30C.
Effect of time on percentage removal of copper using tamarind acid wash sample at 30C
Effect of time on percentage removal of copper using tamarind base wash sample at 30C
Effect of time on percentage removal of copper using carrot distilled water wash sample at 30C
Effect of time on percentage removal of copper using carrot distilled water wash sample at 30C
Effect of time on percentage removal of copper using carrot acid wash sample at 30C
Effect of time on percentage removal of copper using carrot base wash sample at 30C
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Effect of time on percentage removal of copper using carrot base wash sample at 30C
Effect of time on percentage removal of copper using carrot distilled water wash sample at 40C.
Effect of time on percentage removal of copper using carrot distilled water wash sample at 40C.
Effect of time on percentage removal of copper using carrot base wash sample at 40C.
Effect of time on percentage removal of copper using carrot base wash sample at 40C.
Effect of time on percentage removal of copper using tamarind distilled water wash sample at 50C.
SUMMARY AND CONCLUSION

It can be concluded from the laboratory trials both these biosorbent show their potential for commercialization since it is technically feasible, ecofriendly with good metal-binding capacity. It was found that for tamarind distilled water wash and acid wash, initial concentration of 50.5ppm was found to have greater % removal of copper than tamarind base wash. It was observed that tamarind acid wash has more efficient as bioadsorbent with 95% removal of copper.
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It was found that for carrot distilled water wash and acid wash, initial concentration of 100 ppm was found to have greater % removal of copper than tamarind base wash. From this we observed that carrot acid wash has more efficient as bioadsorbent with 96.9% removal of copper. The adsorption capacity was found to be increasing with temperature for both tamarind and carrot biomass. The adsorption capacity was found to be decreasing with increasing pH. Optimum conditions pH 1 there is adsorptivity. biomass 10.5g/l the % removal 5/5/2013 Department of Biotechnology of copper was more. 77
FUTURE PERSPECTIVES
Dynamic studies have to be carried out on different Biosorbents. Surface analysis of the adsorbent. Disorption studies of the adsorbents. Optimising experiment by using software's Batch kinetic study
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References
K.M.S. Sumathi, S. Mahimairaja and R. Naidu, 2004 Use of low-cost biological wastes and vermiculite Solids, for removal of copper from tannery effluent. Bioresource Technology Raji, C. and T.S. Anirudhan, 1997. Copper (II) adsorption by sawdust: kinetics and equilibrium. Indian Journal of Chemical Technology, 4: 228-236. Deans, J.R. and B.G. Dixon, 1992. Uptake of Pb2+ and Cu2+ by novel biopolymers. Water Research, 26(4): 469-472. Veglio, F., F. Beolchini and A. Gasbarro, 1997. Biosorption of toxic metals: an equilibrium study using free cells of Arthrobacter sp. Process Biochemistry., 32: 99-105. Gadd, G.M. and C. White, 1993. Microbial treatment of metal pollution a working biotechnology. Trends Biotechnology, 11: 353-359. Volesky, B., 1987. Biosorbents for metal recovery. Trends Biotechnology, 5: 96-101. Volesky, B. and Z.R. Holan, 1995. Biosorption of heavy metals. Biotechnology Program, 11: 235-250.
ACKNOWLEDGEMENT
We consider it our privilege to express our gratitude and respect to all who guided us in the completion of this project.
We express our sincere gratitude to Dr. Netaji.S.Ganesan, Principal, for providing us with excellent infrastructure to complete our project work. We would like to express our sincere thanks to our HOD, Dr. G.S.Jagannatha Rao for his continuous and never ending support, and without whose help this venture would not be complete. We would like to express our sincere thanks to Dr.Kiran and Dr.Rajeshwari for their valueable suggestions We are deeply indebted to our guide, Dr. Nagamani.S.Khandre and Ms. Shwetha.N, for their timely help, crisp advice, constructive criticism, masterly guidance, academic freedom, unparalleled support, valueable suggestions and for their keen interest in our project. We would like to thank Mr. Sudhanva madhava Desai, Mr.Sinosh skariyachan and Mr. Sameera for their timely help and valueable suggestions while doing our project. Last but not the least we would like to thank the department and all our friends.
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Project Presentation on Optimization of Copper Removal Using Biomass

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Project Presentation

DAYANANDA SAGAR COLLEGE OF ENGINEERING

Under the guidance of Nagamani. S. K, (Guide)

Initial copper ion concentration on the percentage removal of

Temperature on the percentage removal of copper

Practiced in environmental cleanup Heavy metal characteristics

Sources of heavy metals

Metal plating industry

Heavy metal pollution

Effects of Heavy metals

Selenium and Mercury :

Damage to brain, circulatory tissue and central nervous system

Cupric ion : Main ionic form of copper, highly toxic

Physical and chemical properties

Metal adsorption reactions:

The adsorption relationship is expressed as:

log =log K +N log c

R max is the max sorbate uptake under the given conditions

Dissociation of functional group on the cell surface

MATERIALS AND METHODS

Stock solution of copper

Copper reacting with nitric acid

Standard copper solution:

Sodium citrate solution:

Ammonium hydroxide Neocuproine reagent Chloroform

Standardization values and graph

Figures showing the samples prepared for calibration

Tamarind fruit shell (Tamarindus indica)

Carrot peels (Daucus carota)

Batch studies: Following parameters were varied in a

Initial metal ion concentration ( 1ppm 100ppm)

Biomass loading ( 1g/l 20g/l)

Chemicals and Equipments

Specification of materials Department of Biotechnology

POOJA LAB EQUIPMENTS, MUMBAI

RESULTS AND DISCUSSION

Software Minitab version 15

Initial conc (ppm) 1.0 50.5 50.5 100 48.5 50.5

Range of parameters designed using minitab software

Temperature- 30C, 40C, 50C.

Effect of Initial concentration- Tamarind

Ph-4 Temperature- 30C

Biomass loading- 10.5g/l

Series1 93.5 93 92.5 92 91.5 91 0 20 40 60 80 100 120 Initial concentration

Effect of initial concentration- Carrot

Initial conc- 50.5ppm

Biomass loading -20g/l

94 Series1 93.5 93 92.5 92 0 10 20 30 40 50 60 Temperature('C)

Effect of Temperature(Carrot HCl wash)

Ph- 1 Initial conc- 50.5ppm Biomass loading -20g/l

96.45 96.4 96.35 96.3 96.25 96.2 96.15 0 10 20 30 40 50 60 Temperature('C) Series1

Effect of Temperature(Carrot NaOH wash)

Initial conc- 50.5ppm Biomass loading -20g/l

94.8 Series1 94.7 94.6 94.5 94.4 0 10 20 30 40 50 60 Temperature('C)

Effect of Temperature- Tamarind

Ph- 4 Initial conc- 50.5ppm Biomass loading -20g/l

93.5 93 92.5 92 91.5 91 90.5 0 20 40 60 Temperature('C)

Temperature- 40C Initial conc- 100ppm Biomass loading- 10.5g/l

94 93.5 93 92.5 92 91.5 0 2 4 pH 6 8 Series1

Effect of pH on copper adsorption using Tamarind Distilled water

Effect of pH(Tamarind HCl wash)

94 Series1 93.5 93 92.5 92 0 1 2 3 4 pH 5 6 7 8