CHAPTER 1

INTRODUCTION

1.1 GENERAL

Water is a chemical substance that is essential to all known forms of life, and
covers 71% of Earth's surface. It appears that the available water is contained in five
oceans (over 97 %) and the Antarctic ice sheet (2 %). It means that all of the rivers,
streams, lakes, reservoirs, water present as clouds, rain water and freshwater aquifers
constitute only one percent of earth's 'water'. Water in these bodies perpetually moves
through a cycle of evaporation, precipitation and run-off to the sea.
From a biological standpoint, water has many distinct properties that are vital
both as a solvent in which many of the body's solutes dissolve and as an essential part
of many metabolic processes within the body. Water is also central to photosynthesis
and respiration. The civilization has historically flourished around rivers and major
waterways. Mesopotamia, the so-called cradle of civilization, was situated between the
major rivers Tigris and Euphrates. Large metropolises like Rotterdam, London,
Montreal, Paris, New York, Shanghai, Tokyo, and Chicago owe their success in part to
their easy accessibility via water and the resultant expansion of trade. Islands with safe
water ports, like Singapore and Hong Kong, have flourished for the same reason. In
places such as North Africa and the Middle East, where water is more scarce, access to
clean drinking water has been a major factor in human development.
The use of water in the world has increased by more than 35 times over the past
three centuries. Globally 3240 cubic km of fresh water is withdrawn and utilised
annually, and of this 69 per cent is used for agriculture, 23 per cent for industry and 8
per cent for domestic purpose .Increase in human population and industrial growth over
the past 150 years have now caused a serious world-wide water pollution problem. At
present, most of the rivers and lakes of the world are already polluted or at least

1
threatened with pollutants. Human activities are degrading the quality of much more
water than that was withdrawn and consumed.
1.2 ENVIRONMENTAL POLLUTION
Pollution is the effect of undesirable changes in our surroundings that have
harmful effects on plants, animals and human beings. This occurs when only short
term economic gains are made at the cost of long term ecological benefits for
humanity. No phenomenon has led to greater ecological changes than has been made
by man-kind. During the last few decades, we have contaminated our air, water and
land on which life itself depends with a variety of waste products1. Pollution can take
the form of chemical substances or energy, such as noise, heat, or light. Pollutants, the
elements of pollution, can be foreign substances or energies, man-made or naturally
occurring; when naturally occurring, they are considered contaminants when they
exceed natural levels.
1.2.1 Major forms of pollution
1.2.1.1 Air pollution
Air pollution is the release of chemicals and particulates into the atmosphere.
Common gaseous air pollutants include carbon monoxide, sulphur dioxide,
chlorofluorocarbons and nitrogen oxides produced by industry and motor vehicles.
Photochemical ozone and smog are created when nitrogen oxides and hydrocarbons
react with sunlight. Particulate matter, or fine dust is characterized by its micrometre
size 2.5 to 10 pm
1.2.1.2 Light pollution
Light pollution, includes light trespass, over-illumination and astronomical
interference.
1.2.1.3 Noise pollution
Noise pollution, which encompasses roadway noise, aircraft noise, industrial
noise as well as high-intensity sonar.

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1.2.1.4 Soil pollution
Soil contamination occurs when chemicals are released intentionally, by spill or
underground leakage. Among the most significant soil contaminants are hydrocarbons,
heavy metals, herbicides, pesticides and chlorinated hydrocarbons.
1.2.1.5 Radioactive pollution
Radioactive contamination, resulting from 20th century activities in atomic
physics, such as nuclear power generation and nuclear weapons research, manufacture
and devolopment.
1.2.1.6 Thermal pollution
Thermal pollution, is a temperature change in natural water bodies caused by
human influence, such as use of water as coolant in a power plants.
1.2.1.7 Visual pollution
Visual pollution, which can refer to the presence of overhead power lines,
motorway billboards, scarred landforms (as from strip mining), open storage of trash or
municipal solid waste.
1.2.1.8 Water pollution
Water pollution, by the discharge of waste water from commercial and industrial
waste (intentionally or through spills) into surface waters; discharges of untreated
domestic sewage, and chemical contaminants like heavy metals such as lead, mercury,
nickel , copper, cadmium, chromium etc. and textile synthetic dyes like malachite green,
rhodamine -B, methylene blue, congored , crystal violet , acid blue etc.
1.2.2 Pollution by heavy metals
Water is essential for survival. But today about 200 million people in India do not
have access to safe drinking water due to water pollution. Any change in physical,
chemical or biological properties of water is known as water pollution. Heavy metals
and dyes play an important role in water pollution. Heavy metals are continuously
released into the aquatic eco-system from natural process such as volcanic activity and
weathering of rocks. The effluents from mining, ore processing, metal processing, metal
polishing, cleaning, paint manufacturing and battery manufacturing industries and acid
3
rain contribute for the increasing metal loads in the water bodies. The heavy metal
contamination of the water system is responsible for serious diseases and death. Waste
water containing heavy metal pollutants like lead, mercury, nickel, copper, cadmium
and chromium causes direct toxicity, both to human and other living organisms due to
their presence beyond specified limits. The presence of toxic heavy metals in aqueous
streams, arising from the discharge of untreated metal containing effluent into water
bodies, is one of the most important environmental issues. Their presence in aquatic
ecosystem poses serious human health risks and causes harmful effect to living
organisms.
The presence of inorganic pollutants such as metal ion in the ecosystem can cause
a major environmental problem. Toxic metals coming to the earth surface not only
contaminate earth's surface water but it can also affect the subsurface water in trace
amounts by leaking from the soil after rain and snow.

Toxic metal is defined as that metal which is neither essential nor beneficial effect
on the contrary it displays several toxicological symptoms at low levels. These metals
stay almost permanently because they degrade at a very slow rate from the environment.
They pass into the food and they ultimately make their passage into the tissue. They are
not subjected to bacterial attack or other breakdown or degradation process. When they
accumulate in food chains and in the environment, they can profoundly disturb the
biological process.

The primary sources of heavy pollutants in coastal lagoons are input from river
sediments and atmosphere which can affect the aquaculture profitability in certain areas.
The anthropogenic sources of heavy metals include wastes from the electroplating and
metal finishing industries, tannery operations, chemical manufacturing, leather tanning
industries, pigment manufacturing industries, fertilizer industries and battery
manufacturing. The heavy metals which affect the surrounding environment are
Chromium(Cr), copper(Cu), Arsenic(As), Lead( Pb), Nickel(Ni), Cadmium (Cd),
Mercury(Hg), Zinc(Zn), Manganese(Mn).

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1.2.2.1 Chromium

Chromium is one of the toxic contaminants, which exists in hexavalent and

trivalent forms. Hexavalent form is more toxic than trivalent form and requires more
concern. Chromium was regarded with great interest because of its high corrosion
resistance and hardness. A major development was the discovery that steel could be
made highly resistant to corrosion and discoloration by adding chromium to form
stainless steel. The main sources of chromium (Cr6+) are tannery, paint, ink, dye, and
aluminium manufacturing industries. Several International Environmental Agencies
have introduced strict regulations with regard to metal discharge, especially from
industrial activities. According to USEPA, the discharge of Cr6+ and its other forms is
regulated to below 2 mg/ L.

1.2.2.1.1 Applications

The applications of chromium are numerous Chrome plating (electroplating with

chromium) is currently the highest-volume use of the metal. Chromium and
ferrochromium are produced from the single commercially viable ore, chromite, by
silico thermic or alumina thermic reaction or by roasting and leaching processes.
Trivalent chromium (Cr3+) is required in trace amounts for sugar and lipid metabolism.
1.2.2.1.2 Hazards
In larger amounts and different forms, chromium can be toxic and carcinogenic.
Exposure to Cr6+ causes cancer in digestive tract and lungs and may cause epigastric,
pain, nausea, vomiting, severe diarrhoea and haemorrhage. Abandoned chromium
production sites often require environmental clean-up.

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 TABLE 1.1

List of Major Heavy metals and their Health hazards

Contaminants Potential health effects from long term exposure above

maximum contamination level

Arsenic Skin damage or problems with circulatory system

Cadmium Kidney damage
Chromium (Total) Allergic Dermatitis
Copper Gastrointestinal Distress, Liver or kidney Damage
Lead Kidney Damage, High blood pressure
Mercury Kidney problems
1.2.2.2 Lead

Lead is known to be one of the heavy metals and widely used in many industrial
applications. Lead is a soft, malleable metal. Metallic lead has a bluish-white colour
after being freshly cut, but it soon tarnishes to a dull grayish colour when exposed to air.
The permissible limit for Pb (II) in waste water, as given by Bureau of Indian standards
is 0.01mg/L5.
1.2.2.2.1 Applications
Lead is used in building construction, lead-acid batteries, bullets and shots,
weights, as part of solders, pewters, fusible alloys and as a radiation shield. Lead has the
highest atomic number of all of the stable elements, so it is used in electroplating, metal
finishing, textile industries, storage batteries, pigments, as fuel, photographic materials
and explosive manufacturing.
1.2.2.2.2 Hazards
Lead, at certain exposure levels, is a poisonous substance to living organisms. It
damages the nervous system and causes brain disorders. Excessive lead also causes
blood disorders in mammals. Like the element mercury, another heavy metal, lead is a
neurotoxin that accumulates both in soft tissues and in the bones. Lead is highly toxic

6
as its presence in drinking water above the permissible limits causes adverse health
effects such as anaemia, hepatitis, nephritic syndrome and renal damages.
1.2.2.3 Nickel
Nickel is the heavy metal which has been widely used in electroplating industry.
It is a silvery-white lustrous metal with a slight golden tinge. Nickel belongs to the
transition metals and is hard and ductile. Pure nickel shows a significant chemical
activity, though larger collections of the metal are slow to react with air at ambient
conditions due to the formation of a protective oxide film. The permissible limit for Ni
(II) in waste water as given Bureau of Indian standards is 0.02 mg/L.
1.2.2.3.1 Applications
The fraction of global nickel production presently used for various applications
is as follows: 60% for making nickel steels, 14% is used in nickel-copper alloys and
nickel silver, 9% to make malleable nickel, nickel clad, in cone and other super alloys,
6% in plating, 3% for nickel cast irons, 3% in heat and electric resistance alloys such
as chrome, 2% for nickel brasses and bronzes with the remaining 3% of the nickel
consumption is used in all other applications combined. Nickel is used in many specific
and recognizable industrial and consumer products, including stainless steel, alnico
magnets, coinage, rechargeable batteries, electric guitar strings, microphone capsules,
and special alloys.
1.2.2.3.2 Hazards
If waste water contaminated with divalent nickel is discharged to water ways
without adequate treatment, soil and water resources become polluted. Electroplating is
considered a major polluting industry because it discharges toxic materials and heavy
metals through waste water (effluents). It is harmful to humans and other living
organisms. Inhalation of divalent nickel causes an increase in the incidence of lung
cancer. Moreover, soluble nickel compounds are carcinogenic, giving rise to cancers of
the nasal cavities, lungs and other organs such as stomach and kidney. The main
symptoms of nickel poisoning include headache, dizziness, nausea and vomiting, chest

7
pain, tightness of the chest, dry cough and shortness of breath, rapid respiration,
cyanosis and extreme weakness.
1.2.3 Conventional methods for removing Heavy metals
The conventional methods for removing heavy metals from waste waters include
chemical precipitation, electrochemical treatment, evaporation, ion exchange,
membrane process, coagulation and flocculation, adsorption. However, the application
of these treatment processes has been found to be sometimes restricted, because of
expensive investment, operational costs and the potential generation of secondary
pollution. Furthermore such processes may be ineffective or extremely expensive when
the initial heavy metal concentrations are in the range of 10-100 mg/L. In recent years,
bio sorption has been studied extensively using biomass as a bio sorbent for heavy
metals and dyes removal.
1.2.3.1 Chemical Precipitation
This method of treatment is used for the removal of dissolved organic solids.
Lime is usually added as a precipitant in more cases, but more often in combination with
calcium chloride, magnesium chloride, alum, ferric chloride, ferrous sulphate, charcoal
or any one of a number of other substances. Usually ferric chloride is used for the
chemical precipitation.
1.2.3.2 Ion Exchange Process
Ion exchange is a unit process by which an ion of a given species is displaced
from an insoluble exchange material by an ion of a different species in solution. It may
be operated in a batch or continuous mode. In a batch process the resin is simply stirred
with the waste water to be treated in a reactor unit, and the reaction is completed. The
used resins are removed by settling and are regenerated and reused subsequently. In
continuous process, the exchange resins are placed in a bed or a packed column and the
water or waste water to be treated is passed through it.
1.2.3.3 Reverse Osmosis
Reverse osmosis is one of the several demineralisation techniques applicable for
the purification of water or wastewater and make it suitable for usage. Reverse
8
osmosis is a process in which water or wastewater which is separated from dissolved
salts in a solution by filtering through a semi permeable membrane at a pressure
greater than osmotic pressure. Cellulose acetate and nylon have been used as
membrane materials. Reverse osmosis units can be arranged either in parallel to
provide adequate hydraulic capacity or in series to effect the desired degree of
demineralisation.
1.2.3.4 Electro dialysis
In an electrodialysis process, ionic compounds of a solution are separated through
the use of semi-permeable ion-selective membranes. Application of an electric potential
between the two the two electrodes cause an electric current to pass through the solution,
which in turn cause migration of anion towards the positively charge electrode. The
interest in the application has been gaining much interest due to its simplicity and
economy involved in the treatment processes.
1.2.3.5 Membrane filtration
Membrane tlltration technologies with different types of membranes show great
promise for heavy metal removal for their high efficiency, easy operation and space
saving. The membrane processes used to remove metals from the wastewater are
ultrafiltration, reverse osmosis, Nano filtration and electrodialysis.
1.2.3.6 Ultrafiltration
Ultrafiltration is a membrane technique working at low trans-membrane pressures
for the removal of dissolved and colloidal material. Since the pore sizes at UF
membranes arc larger than dissolved metal ions in the form of hydrated ions or as low
molecular weight complexes, these ions would pass easily through ultra-filtration
membranes.
Large molecules and colloidal materials so that this will help to protect the
membranes. The spiral wound configuration of the membranes support structure proves
to be the best and most effective in use when it comes to municipal wastewater
reclamation. However, problems that are still needed to be solved are membrane
durability problems, fouling of membrane, sensitive to hard water salts.
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1.2.3.7 Nanofiltration
Nanofiltration is a membrane filtration-based method that uses nanometer sized
cylindrical through-pores that pass through the membrane at 90°. Nanofiltration
membranes have pore sizes from 1-10 nanometres, smaller than that used in
microfiltration and ultrafiltration, but just larger than that in reverse osmosis.
Membranes used are predominantly created from polymer thin films. Materials that are
commonly use include polyethylene terephthalate or metals such as aluminium. Pore
dimensions are controlled by pH, temperature and time during development with pore
densities ranging from 1 to 106 pores per cm2. Membranes made from polyethylene
terephthalate and other similar materials, are referred to as “track-etch” membranes,
named after the way the pores on the membranes are made. “Tracking” involves
bombarding the polymer thin film with high energy particles. This results in making
tracks that are chemically developed into the membrane, or “etched” into the membrane,
which are the pores. Membranes created from metal such as alumina membranes, are
made by electrochemically growing a thin layer of aluminium oxide from aluminium
metal in an acidic medium.
Nanofiltration is the intermediate process between UF and RO. NF technology
has been used for the rejection of heavy metal ions such as nickel [Murthy et al., 2008],
chromium [Muthu krishnan et al., 2008], copper [C sefalvay et al.. 2004); Ahmad et al.,
2010] and arsenic [Nguyen et al., 2009; Figoli et al., 2010] from wastewater.
1.2.3.8 Coagulation and flocculation
Coagulation and flocculation followed by sedimentation and filtration is also
employed to remove heavy metal from wastewaters. Coagulation is the destabilization
of colloids by neutralizing the forces that keep them apart. Many coagulants are widely
used in the conventional wastewater treatment processes such as aluminum, ferrous
sulfate and ferric chloride, resulting in the effective removal of wastewater particulates
and impurities by charge neutralization of particles and by enmeshment of the impurities
on the formed amorphous metal hydroxide precipitates. General coagulation-
flocculation can't treat the heavy metal wastewater completely. Therefore, coagulation-
10
flocculation must be followed by other treatment techniques. Moreover, this method
consumes chemicals and thus produces large volume of sludge [Fenglian et al, 2011].
1.2.3.9 Biological methods
1.2.3.9.1 Adsorption
Apart from the above mentioned methods, adsorption is highly effective, cheap
and less troublesome. Hence, adsorption by activated carbon is taken as a suitable
method for removal of heavy metals.
Basically, adsorption is a mass transfer process by which a substance is
transferred from the liquid phase to the surface of a solid, and it becomes bound by
physical or chemical interactions (Kurniawan and Babel 2004). Owing to its large
surface area, high adsorption capacity and surface reactivity, adsorption using activated
carbon finds a wide spectrum of application in the removal of metal ions from inorganic
effluent. At the same time the use of commercial activated carbon is also costly and this
keeps the industrialists away from adopting such commercial activated carbon similar
to conventional techniques as discussed earlier.
In recent years, the search for low cost adsorbents that have metal binding
capacities has become intensive. Materials those are locally available in large quantities
such as natural materials or agricultural waste or industrial by-products can be utilised
as low-cost adsorbents. Some of these materials can be directly used as adsorbents
without much effort as involved in conventional approaches. Conversion of these
materials into activated carbon, which can be used as an adsorbent for water purification,
would improve economic value and in a way help industries reduce the cost of waste
disposal and provide a potential alternative to activated carbon.

Bioreduction of metals can be mediated by actively metabolizingbacteria or

through a passive reaction whereby the metal is reduced when it interacts with binding
sites both inside and outside of the cell. Partial metal reductions are usually mediated
by the electron transport systems of the relevant microorganisms, which produce a metal
ion that is less mobile or less toxic in the environment (Groudev and Groudeva 2001).

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 Bioprecipitation of metals occurs when microorganisms produce and secrete
substances that react with soluble metals to yield an insoluble metal compound. Two
essential processes are included in this group: the precipitation of metals as insoluble
metal sulfides as a result of the dissimilatory microbial sulfate reduction and the
precipitation of metal phosphates by enzymatically produced HPO42- (Groudev and
Groudeva 2001).
Bio flocculation uses different microorganisms to facilitate the settling of finely
divided solids in wastewater. In some cases, however, these flocculants are not efficient
or are quite expensive. Bio flocculants may selectively flocculate certain minerals from
slurries containing various mineral constituents (Groudev and Groudeva 2001).
Bioaccumulation is an active process, whereby the biomass transports the metal
ions into the cells where it is then sequestered. The disadvantage of this process is that
the concentration of the metal may reach a level that is toxic to the organism, and kill it
(Volesky 1990).
Bio-methylation and bio-demethylation involve a number of bacteria and fungi
which transform metals by methylation order methylation reactions. Biomethylated
metal derivatives, although toxic, are often volatile and may be eliminated from a system
by evaporation. However, without adequate methods for capture of volatile metals, the
contaminant is merely repartitioned from water, sediments and soils to the atmosphere
(Groudev and Groudeva 2001).

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1.3 OBJECTIVES OF PROJECT

 To determine the amount of Hexavalent Chromium present in the Dye and

Tannery effluent obtained from local industries.
 To study the adsorption characteristics of different naturally available low cost
biosorbents.
 To suggest the best and easily available low cost biosorbent for treatment of
effluent with higher Hexavalent chromium concentration.

13
 CHAPTER 2
LITERATURE REVIEW

2.1 BIOSORPTION
The term ‘‘biosorption’’ is used to describe, in general, the property of microbial
biomass to sequester heavy metals from aqueous systems (Dhami et al 1998).
2.1.1 Mechanism of Biosorption
The active mechanisms of biosorption are complex and often selective (Dhami et
al 1998). They have not been completely elucidated (McAfee et al 2001), are not yet
well understood (Rostami and Joodaki 2002) and still unclear .Biosorption has a
chemical, equilibrated and saturatable mechanism (Dilek et al 2002). Heavy metal
biosorption by microorganisms has been reported to occur via complexation, adsorption
by physical forces (Stirk and van Staden 2002), coordination, chelation, ion exchange,
diffusion through cell walls and membranes (Churchill et al 1995), inorganic
precipitation and/or exchange of counter-ions (Tsezos et al 1995, Kratochvil and
Volesky 1998). They differ depending on the biosorbent species (Zhou and Kiff 1991,
Hafez et al 1997, Kapoor and Viraraghavan 1997), the origin and processing of the
biosorbent and solution chemistry (Ceribasi and Yetis 2001).
2.1.2 Variants of biosorption
In the context of potential exploitation of biosorption in both waste remediation
and resource recovery, the application of physical forces such as magnetism or
electricity have also been documented in literature.
Preparation of non-viable magnetic biosorbents by chemical crosslinking
techniques, making them magnetic and amenable to separation, has been demonstrated
(Patzak et al 1997). According to them, magnetic Saccharomyces cerevisiae had
undiminished silver binding capacity ascompared to native biosorbent; its copper and
cadmium uptake capacity decreased by 40% and 30%, respectively. Biosorption using
electric fields to facilitate contact between a sorbate and non-living biosorbent has been
described by (Riordan et al 2004). (Bustard et al 1998b) have recorded an increase in

14
uranium uptake by spent distillery waste to the tune of 1.5 times when pulse voltage and
capacitance were spiked during electric-field assisted biosorption.
2.1.3 Factors influencing biosorption
Biosorbent concentration influences metal uptake. For instance, (Chandrasekhar
et al 1998) have reported that the uptake of chromium, nickel and iron by Aspergillus
niger increased till 10 g/L, beyond which no change occurred up to 40 g/L. In contrast
to this, (Ruiz-Manriquez et al 1997) have reported decrease in copper biosorption with
increasing concentrations of Thiobacillus ferrooxidans.
(Mohanty and Chaudhury 2002) have reported that biosorption of copper and zinc
by Volvariella volvacea increased with increasing metal concentration. (Mehta et al
2002) have demonstrated that with increase in copper concentration, its biosorption by
Chlorella vulgaris increased rapidly to start with and then gradually up to 100 mM.
Their study also showed that at each concentration of the test metal, the magnitude of
adsorption was higher than that of uptake.
Comparative biosorption of chromium (Cr6+), individually, and in salt containing
environment, by two species of Dunaliella has been demonstrated (Donmez and Aksu
2002). The effect of salinity was studied as a function of NaCl concentration. Their
study showed that with increasing concentrations of salt, chromium uptake by the two
species exhibited a sharp decrease to one-third of the original uptake.
(Goyal et al 2003) have studied the effect of varying the carbon source and
supplementing the nutrients in the biosorbent growth medium. Saccharomyces
cerevisiae grown in glucose or sucrose recorded a greater uptake of chromium (Cr6+)
than those fed with fructose.
Temperature, pH and water hardness have been reported to influence the
biosorption of cadmium and zinc by Fontinalis antipyretica (Martins et al 2004). While
zinc uptake was found to increase with rising temperature and water hardness, cadmium
uptake decreased with the former, being independent of the latter.
Batch experiments on biosorption of Cadmium (II) ions from aqueous solutions
by dead Sargassum sp. were carried out at varying temperature and agitation speed by
15
Cruz et al (2004). Their study revealed decrease in adsorption equilibrium constants
with increasing temperature in the range of 298–328 K, indicating that the process of
Cadmium (II) ions biosorption by Sargassum sp. is exothermic. They have reported
maximum adsorption at agitation speeds greater than 100 rpm.
2.1.4 Biosorption of heavy metals by agriculture wastes and other plant materials:
One of the richest sources for low-cost sorbents is agriculture and especially the
waste resulting from crops. The use of the agricultural products and by-products has
been widely investigated as a replacement for current costly methods of removing heavy
metals from water and wastewater (Kumar, 2006). Due to abundant availability,
agricultural waste poses little economic value and moreover, creates serious disposal
problems (Kumiawan et al., 2006; Tilman et al., 2002). Agricultural materials,
particularly those containing cellulose show potential metal biosorption capacity, so that
diverse agro-waste have been successfully used to remove toxic heavy metals [Pb(II),
Cd(II), Hg(II), Cu(II), Ni(II), Zn(II), Cr(III), and Cr(VI)] from contaminated industrial
and municipal wastewaters (Demirbas, 2008). The types of studied biomasses are
different and depended on the type of agricultural production prevailing in the
geographical areas where the studies were carried out.
The basic components of the agricultural waste materials biomass include
hemicellulose, lignin, extractives, lipids, proteins, simple sugars, water hydrocarbons,
starch containing variety of functional groups that facilitates metal complexation, which
helps for the sequestering of heavy metals (Sud et al., 2008).An analysis of the literature
shows the very wide distribution of countries where the ecological application of
biomasses to metal removal is being developed, and indicates a great interest in
emerging countries such as India, Brazil, Turkey, Argentina and Nigeria (Demirbas,
2008; Kumiawan et al., 2008; Nurchi and Villaescusa, 2008; Sud et al.,2008).
Natural materials or industrial and agricultural wastes are applied in biosorption
technology to remove heavy metals from aqueous media. They offer an efficient and
cost-effective alternative to traditional chemical and physical remediation and
decontamination techniques. Modified agricultural by-product could enhance their
16
natural capacity and add value to the by-product (Kumar, 2006). Biosorbents are
prepared from the naturally abundant or waste biomass either by washing biomass with
acids or bases, or even both, before final drying and granulation. The use of agricultural
residues or industrial by-product as biosorbents have been studied for years, although
the cost of some of these processes proved to be sometimes expensive (Brinza et al.,
2007; Doyurum and Qelik, 2006; Ghodbane et al., 2008; Lesmana et al., 2009; Nouri
et al., 2007). Recently,.application of a number of agricultural materials such as wheat
steams (Tan and Xiao, 2008), Chinese herb Pang Da Hai (Liu et al., 2006), grape bagasse
(Farinella et al., 2008), modified and unmodified oil palm fruit fibre (Abia and Asuquo,
2007), peepul leaves, banana peels, peanut hulls, coir fibers, rice stem, teak saw dust,
discarded tea leaves, mango leaves, rice husk, grass clippings (Gupta et al., 2009),
tobacco dust (Qi and Aldrich, 2008), juniper bark, wood (Nouri et al., 2007), neem leaf
powder (Sharma and Bhattacharya, 2008), pine bark (Gundogdu et al., 2009), bael
leaves (Chakravarty et al, 2010), Moringa oleifera bark (Reddy at el, 2010) etc., have
been reported for the removal of different heavy metals from aqueous solution.
Most of the studies showed that the heavy metal biosorption by agricultural waste
materials and by-products are quite high and varies from 50 to 100%. Biosorbent used
in this discussion were focused only on biosorption of Pb(II), Cd(II), Cu(II) and Cr(VI)
ions as these are the metals used in this study.
2.2 BIOSORPTION OF HEAVY METALS BY MICROORGANISMS
A large number of microorganisms belonging to various groups, viz. bacteria,
fungi, yeasts, cyanobacteria and algae have been reported to bind a variety of heavy
metals to different extents. The role of various microorganisms by biosorption in the
removal and recovery of heavy metal has been well reviewed and documented (Stratton,
1987; Gadd and Griffiths, 1978; Volesky, 1990; Wase and Foster, 1997; Gadd 1988;
Mehta et al., 2005; Romera et al, 2006). Most of the biosorption studies reported in
literatures have been carried out with living microorganisms. However due to certain
inherent disadvantages, use of living microorganisms for metal removal and recovery is
not generally feasible in all situations. For example, industrial effluents contain high
17
concentrations of toxic metals under widely varying pH conditions. These conditions
are not always conducive to the growth and maintenance of an active microbial
population. The microbial processes are inexpensive and pollution free. However, the
technique is relatively new and requires special skill in handling the microbe strains for
treating the metal contaminated solution. Since long-term impacts of most of the strains
are not yet known, many are skeptical and advocate further intensive studies before
employing such microorganisms in environmental processes.
Biosorption of metal ions onto microorganisms involve a combination of the
metal-binding mechanisms including physical adsorption, ion exchange, complexation
and precipitation (Ahalya et al., 2003). A variety of ligands is located on the fungal cell
walls is known to be involved in metal chelation. These include carboxyl, amine,
hydroxyl, phosphonic and sulphonic groups (Le Cloirec et al., 2003). Metal ions could
be adsorbed by complexing with negatively charged reactions sites on the cell surface.
These tables showed that both active (living) and inactive (dead) cells of each
microorganisms have been tested to adsorb Pb(II), Cd(II), Cu(II) and Cr(VI) from
aqueous solution. Biosorbent used in these tables were focused only on Pb(II), Cd(II),
Cu(II) and Cr(VI) ions removal as these are the metals used in this study.
2.3 REMOVAL OF HEXAVALENT CHROMIUM USING ORGANIC
ADSORBENTS
Ayub et al (2003) evaluated Chromium removal by Adsorption on coconut Shell.
The effect of pH, contact time, adsorbent dose, and concentration of metal, particle size
and temperature were studied. The coconut shell exhibits good adsorption
characteristics and the data follow both Freundlich and Longmuir models.
Babu et al (2008) investigated that adsorption of Cr (VI) using activated neem
leaves using kinetic studies. The adsorption of Cr (VI) is found to be maximum (99 %)
at low values of pH in the range of 1 – 3. A small amount of the neem leaves adsorbent
(10 g/L) could remove as much as 99 % of Cr (VI) from a solution of initial
concentration 50 mg/L. The adsorption process of Cr (VI) is tested with Langmuir
isotherm model.
18
 Baisakh and Patnaik (2002) studied “Removal of Hexavalent Chromium from
Aqueous Solutions by Adsorption on Coal Char. The effects of pH, temperature and
concentration of hexavalent Chromium were investigated. Under ambient temperature
and pH condition removal efficiency to the extent of 70 %. The rate process follows first
order kinetics.
Bishnoi et al (2003) conducted a study on Cr (VI) removal by rice husk activated
carbon from aqueous solution. They found out that the maximum metal removal by rice
husk took place at pH 2.0 and multilayer adsorption might occur on the surface of the
adsorbent, as indicated by the applicability of the Freundlich isotherm for the
equilibrium data.
Emine Malkoc et al (2006) conducted batch and column studies to find the
adsorption capacity of Chromium (VI) on pomale an olive oil indent waste. The effect
of pH on the Cr (VI) was investigated by varying the pH value as 2, 3, 4 and 5 by fixing
the contact time as 180 min. The adsorption capacity of Cr (Vl) at pH 2 was 8.4mg/g
which reduced to 2.7 mg/g at pH 5. In the column study influent flow rate were varied
from 5 mL/min to 20 mL/min with bed depth as 10 cm. It was found that breakthrough
curve becomes steeper when the flow rate was increased.
Hameed et al (2009) have studied the effect of pH on the adsorbing capacity of
dye using agricultural waste. pH was varied from 2 to 10 and it was found that the
adsorbing capacity increased from 13.29 mg/g to 104.50 mg/g when the pH varied
between 2 and 10 .The authors used pineapple stem for removing cationic dye and
studied the potentiality of its usage. The surface texture of the adsorbent was studied
using Scanning Electron Microscope before and after adsorption and Fourier Transform
Infra-Red analysis was applied and the spectra was recorded. From the study, the results
indicate that pineapple stem was very effective in its adsorbent property.
Kermit Wilson et al (2006) have carried out research on select metal adsorption
by using activated carbon made from peanut shell. The metals considered were
cadmium, copper, lead, nickel and zinc. The activated carbon was prepared by different
methods such as pyrolysis steam activation, oxidation and ash removal. It was found
19
that carbons with higher titratable functional groups are better at binding metal ions than
those carbons with fewer amounts of titratable functional groups.
Li et al (2009) studied the feasibility of preparation of activated carbon from
coconut shell chars in pilot – scale microwave heating equipment. Experiments
conducted to prepare activated carbon by microwave heating indicated that microwave
energy decreases the reaction temperature, saving the energy and shortening processing
time remarkably compared to the conventional heating.
Manjeet Bansal et al (2008) studied the removal of Cr (VI) from aqueous solution
using rice husk. He used two forms of rice husk namely boiled rice husk and
formaldehyde treated rice husk. He conducted batch studies by varying the parameters
such as pH, dosage and agitation speed. The initial concentration of the solution was
taken as 100mg/L, the contact time was fixed as 180 min, pH of the solution was
maintained as 2. The removal efficiency for boiled rice husk was found to increase from
33.2% to 71% with increase in dosage from 2 g/L to 4 g/L. Similarly for formaldehyde
treated rice husk the removal efficiency increased from 38% to 78.5% with the increase
in dosage from 5 g/L to 4 g/L. The results indicate that the metal removal was found to
be maximum at pH 2. It is also found that there is no much difference in adsorption
capacity of boiled rice husk and formaldehyde treated rice husk.
Olayinka Kehinde et al (2009) have compared the efficiencies of two low cost
adsorbents in the removal of chromium and nickel from aqueous solution. Coconut husk
and teak tree bark based adsorbents were employed for the removal of chromium and
nickel. The influence of pH, dosage, contact time and temperature were studied. The
coconut husk adsorbents gave better results the teak tree bark adsorbents at the increased
contact time it was also found that the percentage of adsorption of the adsorbents
increased with increasing adsorbent dosage. From the results it is found that coconut
husk adsorbent was more efficient in removing the metals when compared with teak tree
bark adsorbent.
Rakhi Gopalan (2007) has investigated the adsorption of hexavalent Chromium
from the chrome plating industry effluent using tapioca peel carbon. Both batch and
20
column experiments were conducted and the optimum pH and dosage were found to be
3 and 400 mg respectively. The removal efficiency was found to be 82% based on the
experiments. They have concluded that tapioca peel carbon is more effective in
removing Chromium from wastewater.
Ramesh et al (2009) have prepared activated carbon from green pea peels for
treating dyeing industry wastewater. Wastewaters from textile industry were collected.
100 ml of the sample was collected and agitated with 0.5 g of adsorbent at 120 rpm. The
adsorbent dose was varied from 0.5 g to 3.5 g and pH was varied from 3 to 10. The
results revealed that the removal efficiency increases with agitation time and attains
equilibrium after 60 minutes and remains constant thereafter.
Rao et al (2003) have conducted experiments on removal of Cr(VI) and Ni(II)
using low – cost adsorbents. In this study, the efficiency of low cost materials, namely
bagasee and fly ash were assessed. These low cost adsorbents are used for the removal
of Chromium and Nickel (II) from aqua solution. The effects of pH, contact time, initial
concentration of adsorbate and particle size on the uptake of Chromium and Nickel (II)
have been studied in the experiments. The order of selectivity is PAC > bagasee > fly
ash for Cr (VI) removal and PAC > fly ash > bagasee for Ni (II) removal at optimum
conditions.
Samantaroy et al (1998) made a comparative study on the removal of chromium
(VI) by different adsorbents namely Kendu fruit Gum Dust, Drumstick Gum Dust and
Green Peas Skin Dust. It was found that KGD is the most effective one and concluded
that the removal of Cr (VI) upto 100% was observed with KGD, DGD and GPSD.
Rai and Surendrakumar (1999) conducted experiments on Treatment of
Chromium bearing wastewater by adsorption on brick kiln and fly ash. Suitability of
brick kiln ash and fly ash for removing Chromium (VI) exhibited good sorption at a pH
of 1.3 and particle size 124 – 853 μm. The author reported that fly ash adsorbent was
more effective than brick kiln ash due to smaller particle size.
Singh et al (2001) studied the removal of Chromium (VI), Iron (III) and mercury
(II) from aqueous solutions using activated carbon obtained from used Tea leaves. Break
21
through and exhaustive capacities, removal and recovery of Cr (VI), Hg (II) and Fe (III)
and effect of diverse ions have been studied. The experimental results showed that Cr
(VI), Fe (III) and Hg (II) were strongly adsorbed by the adsorbent; Zi (II), Mn (II) and
Cu (II) were adsorbed partially.
Sreenivasulu et al (2010), have conducted experiments on the adsorption of
chromium on acid treated soapnut tree stem bark carbon. The concentration of
chromium was varied from 1.25 mg/L to 50 mg/L. The stirring speed was maintained
as 120 rpm. Equilibrium time was maintained at 5 hours. The experiment data show
68.4% removal for chromium ion at pH 3.7 and adsorbent dose of 6 g/L.
Subramanyan Vasudevan et al (2009) have carried out a research on the removal
of iron from drinking water by electro coagulation. The parameters such as pH,
temperature and current density were considered. Langmuir and Freundlich isotherms
were plotted. The results revealed that the removal efficiency of 98.8% was achieved at
pH 6.5.The temperature studies also showed that the adsorption was endothermic and
spontaneous.
Tonni Agustiono Kurniawan et al (2006) compared the effects of low cost
adsorbents for treating waste waters laden with heavy metals, it was found that the low
cost adsorbents prepared from agricultural waste have demonstrated outstanding
removal capacity for Cr (VI). The adsorption capacity was 170 mg/g of hazelnut shell
activated carbon, for Ni (II), it was 158mg/g of orange peel, for Cu (II), it was 154.9mg/g
of chemically modified soybean hull, for Cu (II), it was 52.08 mg/g of jackfruit.
Vishwanath (1974) has investigated the effect of Lignite in the removal of Nickel
from the aqueous solution and conducted batch experiments. The effect of the
parameters such as sorbent particle size, pH sorbent/sorbent ratio, retention time,
temperature and carbonate ion concentration was varied. The removal efficiency was
found to be 98% at the retention time of 30 minutes and the optimum pH was found to
be 12.6.
Wang et al (2008) investigated the removal of Cr (VI) from aqueous solutions
using Alligator weed, a freshwater macrophyte, was investigated in batch studies.
22
Various factors including solution pH, Cr (VI) concentrations, agitation time, and
temperature were taken into account and promising results obtained. An initial solution
pH of 1.0 was most favourable for Cr (VI) removal. The results suggested that the Cr
(VI) adsorption at all temperatures was best represented by the pseudo-second-order
equation.

23
 CHAPTER 3

MATERIALS AND METHODS

3.1 CHARACTERISTICS OF TANNERY WASTEWATER

Tannery wastewater contains large amount of chemical compounds including

toxic substances. The major public concern over tanneries has traditionally been about
odours and water pollution from untreated discharges. Important pollutants associated
with the tanning industry include chlorides, tannins, chromium, sulphate and sulphides
as addition to trace organic chemicals and increasing use of synthetic chemicals such as
pesticides, dyes and finishing agents, as well as from the use of newer processing
chemical solvents. These substances are frequently toxic and persistent, and affect both
human health and the environment. Tannery effluent is among one of the hazardous
pollutants of industry. Major problems are due to wastewater containing heavy metals,
toxic chemicals, chloride, lime with high dissolved and suspended salt and other
pollutants. Bioremediation technology to treat hazardous waste has gained considerable
attention as it is ecologically sound and economical relative to other technologies and it
has been used successfully in many countries of the world

Many conventional processes were carried out to treat wastewater from tannery
industry such as biological process, oxidation process and chemical process etc. Among
these, physical and chemical methods are considered very expensive in terms of energy
and reagents consumption. Various physiochemical techniques used for wastewater
treatment can be applied to tannery wastewater (to the entire process or to individual
step in the process) but these processes are expensive. Biological treatment of
wastewater is more favorable and cost effective as compared to other physiochemical
methods. Various microorganisms are capable of reducing the content of pollutants
significantly by utilizing them as energy and nutrient source in the presence or absence
of oxygen.

24
 FIG 3.1 Tannery effluent obtained from a Tannery in Arcot

TABLE 3.1

General Standards of Heavy metal for Waste water disposal as per CPCB

S. No. Parameter Standards

Inland Public Land for Marine coastal areas
surface sewers irrigation
water
1 2 3
A B C d
1 Arsenic as As 0.2 0.2 0.2 0.2
mg/l, max.
2 Mercury (as 0.01 0.01 -- 0.01
Hg) mg/l,
max.
3 Lead (as Pb) 0.1 1.0 -- 0.01
mg/l, max
4 Cadmium (as 2.0 1.0 -- 2.0
Cd) mg/l,
max.
5 Hexavalent 0.1 2.0 -- 1.0
chromium
(as Cr+6),
mg/l max.
6 Total 2.0 2.0 -- 2.0
chromium
(as Cr) mg/l,
max
25
7 Copper (as 3.0 3.0 -- 3.0
Cu) mg/l,
max.
8 Zinc (as Zn) 5.0 15 -- 15
mg/l, max
9 Selenium ( as 0.05 0.05 -- 0.05
Se) mg/l, max
10 Nickel (as Ni) 3.0 3.0 -- 5.0
mg/l, max.
11 Manganese 2 mg/l 2 mg/l -- 2 mg/l
(as Mn)
12 Iron (as Fe) 3 mg/l 3 mg/l -- 3 mg/l
13 Vanadium (as 0.2 0.2 -- 0.2 mg/l
V) mg/l mg/l

TABLE 3.2

Discharge Standards specific to Dye Industry as per CPCB

Parameter Concentration not to exceed,

milligram per litre (mg/l),
except pH

pH 5.5 – 9.0

Total suspended solids 100

Bio-chemical oxygen demand

30
(BOD)

Chemical oxygen demand (COD) 250

Total residual chlorine 1

Oil and grease 10

Total chromium as Cr 2

Sulphide as S 2
TABLE 3.3

26
 Discharge Standards specific to Tannery Industry as per CPCB

S. Effluent Parameter Concentration

No.
1 Ph 6.5 – 9.0
2 BOD (3 days at 27 C) 100 mg/l
3 Suspended solids 100 mg/l
4 Sulphides as S 1 mg/l
5 Total Chromium as Cr 2 mg/l
6 Oil and Grease 10 mg/l
7 Waste water Generation 28 m3/T

3.2 MATERIALS USED FOR BIOSORPTION

3.2.1 Fish scales:

Biosorption of heavy metals by Mozambique tilapia (M. tilapia) fish scales is one
of the treatment that has emerged as an environmental friendly method for the removal
of metal from synthetic and domestic wastewater. The objectives of this study the
efficiency of fish scale in removing chromium ions wastewater. This study highlighted
that fish scale is a promising adsorbent in removing chromium ion from synthetic and
domestic wastewater solution. Seashell, mandarin peels, rice bran, crab carapace, pecan
nutshell, algae-yeast, peanut shell, palm shell, and others have been discovered as a
natural absorbent to remove heavy metals.

Recently, studies had reported the used of fish scales. The cell wall surface of fish
scale contained several of functional group such as carbonyl, nitro, and amine groups
for metal ions attached onto fish scale and the porous layer may provide a good
possibility of metal ions to be adsorbed on its surface. Therefore, this research will
explore the unexploited property of local fish scales, available in abundance as waste

27
material from the wet market, as a new biosorptive approach to remove target heavy
metals such as chromium ions from wastewater.

3.2.1.1 Preparation of biosorbent

Waste fish scales fresh water fishes like Catla were obtained from the fish market.
The fish scales were soaked in water and stored in a thick plastic container to remove
any adhering dust and soluble impurity. Then the fish scales are washed and sun-dried.
The fish scales are used without powdering the process.

3.2.2 Banana peels.

Banana peel, which is a discarded fruit waste and which are readily available
waste in the market and are used to prepare environment benign bio adsorbent for the
adsorption of impurities from aqueous solution. Banana peels were washed, dried and
shredded before being used for treatment of waste water.

Banana peel seems to be good adsorbent and can be used as valuable material for
cleaning of water. Bio adsorbent prepared from banana peels has been reported for the
removal of chromium, cadmium and copper ions from aqueous solution.

Fresh banana peels were collected from domestic wastes, as its availability and
transportation was easy. Banana peel contains lipids (1.7%), proteins (0.9%), crude fiber
(31%) and carbohydrates (59%). The various minerals present are potassium (78.10
mg/g), manganese (76.20 mg/g), sodium (24.30 mg/g), calcium (19.20 mg/g) and iron
(0.61 mg/g). The peels were washed several times with tap water and followed by
distilled water. The washed material then cut in to small pieces and allowed to dry in a
hot air oven at 80˚C for 24 hours. The moisture content was lost from it and the color
change was observed from yellow to brownish black. The dried material was finely
ground and shredded.

28
3.2.2.1 Preparation of biosorbent

The peels were washed several times with tap water, followed by distilled water.
The washed material was then cut in to small pieces and allowed to dry in a hot air oven
at 80˚C for 24 hours. The moisture content was lost after oven drying and the color
change was observed from yellow to brownish black. The dried material was finely
ground and shredded.

3.2.3 Rice Husk

Rice husk is a by-product of the rice milling industry. Rice husk is commonly
used as a low-value energy resource, burned in the field, and/or discarded into the
environment. Rice husk mainly consists of ash and organic matter, which comprises of
organic cellulose, hemi-cellulose, lignin and inorganic silicon dioxide. Rice husk is
composed of 20 % ash, 32.2-38 % cellulose, 21.3- 22 % lignin 21.4 % of hemi-cellose,
18 % pentose, and 2 % of other organic components. As rice husk is insoluble in water,
it has good chemical stability, structural strength due to the high silica content.

3.2.3.1 Preparation of Biosorbent

A homogeneous mixture of rice husk was thoroughly washed with tap water to
clean dirt and mud. The cleaned rice husk was allowed to dry in sunlight for 24 hours
for the removal of moisture. Then the rice husk is blended with blender to form a fine
powder.
3.2.4 Rice Husk Ash (RHA)

RHA is light in weight and has approximately 15-20 % of paddy weight. Rice
husk ash is produced from the combustion of rice husk. The colour of RHA varies from
white to grey. Rice husk is commonly used as a low-value energy resource, burned in
the field, and or discarded into the environment. Rice husk mainly consists of ash and
organic matter, which comprises an organic cellulose, hemi-cellulose, lignin and
inorganic silicon dioxide. Further, reported that rice husk is composed of 20 % ash, 32.2-
38 % cellulose, 21.3- 22 % lignin, 21.4 % hemi-cellulose, 18 % pentose, and 2 % of
29
other organic components. As rice husk is insoluble in water, it has good chemical
stability and structural strength due to the high silica content. RHA, produced after the
burning of Rice husks (RH) has high reactivity and pozzolanic property. Rice husk ash
contains nearly 90-96 % of silica which may be used as one of the ingredients for making
ceramic, polymers and electronic products, it has highly porous and light in weight, with
a very high external surface area.
3.2.5 Crown flower

The bio sorption of chromium heavy metal ions on Calotropis procera (Crown
flower) appears to be an efficient and low cost alternative to be considered in industrial
wastewater treatment. Calotropis procera, also called rooster tree, giant milkweed or
Sodom apple, is a member of flowering plant species in the dogbane family,
Asclepiadaceae. This plant is a soft wooded, evergreen, perennial shrub. Giant
milkweed is native to West Africa as far south as Angola, North and East Africa and
south Asia. Calotropis procera which is known to be locally used in coagulation of milk
to cheese. This implies that it might have some abilities to coagulate or adsorb metallic
macromolecules and therefore expected to sorb heavy metals from aqueous solutions
containing them.

3.2.5.1 Preparation of Biosorbent

Calotropis procera (Crown Flower) is available in many places such as banks of

lakes, roadways, barrel land etc. This flower is collected from lands in and around our
hostel. The collected flowers are sun dried and the flowers are blended in blender to
form a fine powder.

3.2.6 Banana stalk

Banana Stalks (BS) are generated in large amount as waste product after the
consumption of banana and these are left to decompose being of non-economic
importance . However, Banana Stalk is a rich lignocellulosic agricultural waste, which
can be used to recover energy as an adsorbent.

30
 FIG 3.2 Fish scales (Washed and
FIG 3.3 Banana peels (Shredded)
Sun-dried)

FIG 3.4 Rice husk (Powdered) FIG 3.5 Rice husk ash

FIG 3.6 Crown flower (powdered) FIG 3.7 Banana stalk (Shredded)

31
3.2.6.1 Preparation of Biosorbent

Fresh and green Banana stalks are collected and dried in sunlight. The dried
material is cut into small pieces and shredded using blender.
3.3 METHOD OF CHROMIUM DETERMINATION

Chromium is found chiefly in chome-iron ore. Chromium is used in alloys, in

electroplating and in pigment. Chromium salts are used extensively in industrial
processes and may enter a water supply through the discharge of wastes. Chromate
compounds frequently are added to cooling water for corrosion control. Chromium may
exist in water supplies in both the hexavalent and the trivalents state although the
trivalent form rarely occurs in potable water. FAO recommended the maximum limit
for irrigation water as 100µg/L. USEPA drinking water standard for chromium is
0.1mg/L.

3.3.1 Methods for analysis:

A. Inductively coupled plasma method/Atomic absorption spectrophotometer

B. Colorimetric method

Colorimetric method has been followed to find the Hexavalent Chromium(Cr6+)

concentration and has been discussed below

3.3.1.1 Colorimetric method

Principle:

This procedure measures only hexavalent chromium (Cr6+). Therefore, to

determine total chromium convert all chromium to the hexavalent state by oxidation
with potassium permanganate. The hexavalent chromium is determined colorimetrically
by reaction with diphenylcarbazide in acid solution. A red-violet colour of unknown
composition is produced which is measured at 540 nm. To determine total chromium,
digest the sample with a sulphuric-nitric acid mixture and then oxidise with potassium
permanganate before reacting with the diphenylcarbazide.
32
Interferences:

The reaction with diphenylcarbazide is nearly specific for chromium. Hexavalent

molybdenum and mercury salts will react to form colour with the reagent but the
intensities are much lower than that for chromium at specified pH. Vanadium interferes
strongly but concentrations up to 10 times that of chromium will not cause trouble. Iron
in concentrations greater than 1mg/L may produce a yellow colour but the ferric ion
(Fe3+) colour is not strong and no difficulty is encountered normally.

Apparatus and equipment

i. Spectrophotometer: for use at 540 nm, with a light path of 1cm.

Reagents and standards

Use reagents and distilled water free from Chromium contamination.

i. Stock chromium solution: Dissolve 141.4 mg K2Cr2O7 in water and dilute to

100mL; 1mL = 500µg Cr on curve.
ii. Standard chromium solution: Dilute 1mL stock chromium solution to 100mL;
1mL = 5µg Cr.
iii. Nitric acid: HNO3, conc.
iv. Sulphuric acid: H2SO4, conc.; H2SO4, 1+1 (18N); H2SO4 (6N)
v. Sulphuric acid: H2SO4, 0.2N: Dilute 17mL 6N H2SO4 to 500mL with water.
vi. Phosphoric acid: H3PO4, conc.
vii. Diphenylcarbazide solution: Dissolve 250mg 1, 5-diphenylcarbazide (1,5-
diphenylcarbohydrazide) in 50 mL acetone. Store in a brown bottle. Discard
when solution becomes discoloured.
viii. Sodium hydroxide 1N: Dissolve 40g NaOH 1L water. Store in plastic bottle.

33
Procedure

Preparation of calibration curve:

To compensate for possible slight losses of chromium during analytical

operations, treat chromium standards by the same procedure as the sample. Accordingly,
pipette measured volumes of standard chromium solution (5µg/L) ranging from 2 to
20mL, to give standards for 10 to 100 µg Cr, into 250mL beakers or conical flasks.

Colour development and measurements:

Add 0.25mL (5 drops) H3PO4. Use 0.2N H2SO4 and pH meter to adjust solution
to pH ± 0.5. Transfer solution to a 100mL volumetric flask, dilute to 100mL and mix.
Add 2mL diphenyl carbazide solution, mix and let stand 5 to 10 min for full colour
development. Transfer an appropriate portion to 1 cm absorption cell and measure its
absorbance at 540nm, using reagent water as reference. Correct absorbance of a blank
carried through the method. From the corrected absorbance, determine micrograms
chromium present by reference to the calibration curve.

Construct a calibration curve by plotting corrected absorbance values against

micrograms chromium in 102mL final volume. Repeat the experiment with samples.

3.4 METHOD OF EFFLUENT TREATMENT USING BIOSORBENTS

A calibration curve is needed to determine the amount of chromium. Hence, 6
standard solution of Hexavalent Chromium is prepared of concentrations 10 ppm, 5
ppm, 1 ppm, 0.5 ppm, 0.2 ppm and 0.1 ppm. For the color development, orthophosphoric
acid and diphenyl carbazide solution are added to the standards. The absorbance for
each standards at 540 nm is noted down. A calibration curve is plotted with Absorbance
in X axis and Concentration in Y axis. From this calibration curve, the Hexavalent
chromium present in the sample can be found out.

34
 FIG 3.8 Color development in Standard solution of 10 ppm
The collected effluent sample is filled in 6 Nos. 500 mL beakers. The sample
maybe either diluted or not depending on the requirement. A small amount of effluent
sample is taken in the beaker and added with 2mL of Ortho Phosphoric acid and 2mL
of 1,5 – diphenyl carbazide. Now the sample is allowed to stand for 5 – 10 minutes. A
reddish violet color development is noted in the sample. Now a small amount of this
sample is taken in the 1 cm sample cell and measured for absorbance at 540 nm
wavelength in the UV-Vis spectrophotometer. The concentration corresponding to the
absorbance gives the concentration of hexavalent chromium present in them.

FIG 3.9 UV Visible Spectrophotometer

35
 Now, the effluent in beakers are added with dosage of the one of the biosorbents
as 2.5 mg/l, 5 mg/l and 10mg/l for the first three beakers and with constant dosage of 10
mg/l for the remaining 3 beakers. The time at which the biosorbents are added is noted
down. Then for the corresponding time, the effluent with biosorbents are filtered and
collected in a separate beaker. Now, the same procedure for determining the chromium
concentration using absorbance is adopted to determine the amount of Chromium
present in the treated sample effluent.

FIG 3.10 Effluent sample with Shredded Banana peels

36
 CHAPTER 4

RESULTS AND DISCUSSION

4.1 TREATMENT OF TANNERY EFFLUENT USING BIOSORBENTS

The effluent is treated with each biosorbent for varying period of time for constant
dosage and for varying dosage for a constant period of time. The Trivalent chromium is
necessary for all living organisms as a dietary nutrient. It is required to potentiate insulin
and for normal glucose metabolism. Cr (III) is found in most fresh foods and drinking
water. At the same time, the hexavalent Chromium is highly carcinogenic and occurs
rarely by natural methods and is mostly due to man-made activities. This hexavalent
chromium can cause skin irritations, nose irritations and lung cancer if exposed. Hence,
removal of these hexavalent chromium present in the industrial effluent becomes vital.

4.2 RESULTS

4.2.1 Preparation of Calibration curve

TABLE 4.1

Calibration Curve readings

S. Concentration of Absorbance
No. Standard solution
(ppm)
1 10 3.365
2 5 2.08
3 1 0.51
4 0.5 0.135
5 0.2 0.095
6 0.1 0.042

37
A graph is plotted with Absorbance in X axis and Concentration in Y axis using
Origin Pro software.

FIG 4.1 Calibration curve

4.2.2 Concentration of chromium in untreated sample

The calibration curve so obtained is used to determine the chromium present in

effluent sample. It is determined by using origin pro data analysis software.

TABLE 4.2

Concentration of Cr6+ in samples

S. Effluent sample Absorbance Concentration of Hexavalent Chromium

No. from calibration curve (ppm)
1 Dye 0.045 0.012
2 Tannery 1.848 5.138

38
 Since the Hexavalent chromium content in Dye effluent is very much lower than
the discharge standards, tannery effluent is used to determine the adsorption capacity of
the biosorbents.

4.2.3 EFFLUENT TREATED USING DIFFERENT BIOSORBENTS

4.2.3.1 RICE HUSK

TABLE 4.3

Variation of Dosage of Rice husk

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
2.5 1 0.414 4.168 18.42
5 1 0.384 3.824 25.15
10 1 0.305 2.92 42.85

Efficiency
45

40

35

30
Efficiency %

25

20

15

10

0
0 2 4 6 8 10 12
Dosage g/l

Fig 4.2 Efficiency Vs Dosage of Rice Husk Powder

39
 TABLE 4.4

Variation of Time of Contact of Rice husk

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
10 0.5 0.409 4.108 19.59
10 1 0.371 3.676 28.05
10 3 0.234 2.108 58.74

Efficiency Vs Time of Contact

70

60

50
Efficiency %

40

30

20

10

0
0 0.5 1 1.5 2 2.5 3 3.5
Time of Contact (hour)

Fig 4.3 Efficiency Vs Time of Contact of Rice Husk powder

40
4.2.3.2 CROWN FLOWER

TABLE 4.5

Variation of Dosage of Crown flower powder

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
2.5 1 0.448 4.556 10.82
5 1 0.349 3.424 32.98
10 1 0.276 2.588 49.34

Efficiency Vs Dosage
60

50

40
Efficiency %

30

20

10

0
0 2 4 6 8 10 12
Dosage g/l

Fig 4.4 Efficiency Vs Dosage of Crown flower powder

41
 TABLE 4.6

Variation of Time of Contact of Crown flower powder

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
10 0.5 0.430 4.348 14.90
10 1 0.498 4.04 20.92
10 3 0.323 3.128 38.77

Efficiency Vs Time of Contact

45

40

35

30
Efficiency %

25

20

15

10

0
0 0.5 1 1.5 2 2.5 3 3.5
Time of Contact hr

Fig 4.5 Efficiency Vs Time of Contact of Crown flower powder

42
4.2.3.3 RICE HUSK ASH

TABLE 4.7

Variation of Dosage of RHA

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
2.5 1 0.223 1.984 61.17
5 1 0.217 1.916 62.50
10 1 0.208 1.812 64.53

Efficiency Vs Dosage
65

64.5

64

63.5
Efficiency %

63

62.5

62

61.5

61
0 2 4 6 8 10 12
Dosage g/l

Fig 4.6 Efficiency Vs Dosage of RHA

43
 TABLE 4.8

Variation of Time of Contact of RHA

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
10 0.5 0.302 2.236 43.47
10 1 0.254 2.34 54.20
10 3 0.245 2.888 56.23

Efficiency Vs Time of Contact

60

50

40
Efficiency %

30

20

10

0
0 0.5 1 1.5 2 2.5 3 3.5
Time of Contact hr

Fig 4.7 Efficiency Vs Time of Contact of RHA

44
4.2.3.4 SHREDDED BANANA STALK

TABLE 4.9

Variation of Dosage of Banana stalk

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
2.5 1 0.466 2.38 53.42
5 1 0.428 2.164 57.64
10 1 0.424 2.14 58.11

Efficiency Vs Dosage
59

58

57
Efficiency %

56

55

54

53
0 2 4 6 8 10 12
Dosage g/l

Fig 4.8 Efficiency Vs Dosage of Banana stalk

45
 TABLE 4.10

Variation of Time of Contact of Banana stalk

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
10 0.5 0.433 2.192 57.10
10 1 0.404 2.026 60.34
10 3 0.397 1.986 61.13

Efficiency Vs Time of Contact

61.5
61
60.5
60
Efficiency %

59.5
59
58.5
58
57.5
57
56.5
0 0.5 1 1.5 2 2.5 3 3.5
Time of Contact hr

Fig 4.9 Efficiency Vs Time of contact of Banana stalk

46
4.2.3.5 FISH SCALES

TABLE 4.11

Variation of Dosage of Fish scales

Amount Time Absorbance Chromium Removal efficiency

of adsorbent duration concentration %
(g/l) (hour) (ppm)
2.5 1 0.884 2.384 53.34
5 1 0.860 2.316 54.67
10 1 0.834 2.241 56.14

Efficiency Vs Dosage

56.5

56

55.5
Efficiency %

55

54.5

54

53.5

53
0 2 4 6 8 10 12
Dosage g/l

Fig 4.10 Efficiency Vs Dosage of Fish scales

47
 TABLE 4.12

Variation of Time of Contact of Fish scales

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
10 0.5 0.948 2.527 49.76
10 1 0.895 2.375 52.73
10 3 0.624 1.597 67.88

Efficiency Vs Time of contact

80

70

60

50
Efficiency %

40

30

20

10

0
0 0.5 1 1.5 2 2.5 3 3.5
Time of Contact hr

Fig 4.11 Efficiency Vs Time of Contact of Fish scales

48
4.2.3.6 BANANA PEELS (SHREDDED)

TABLE 4.13

Variation of Dosage of Banana peels

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
2.5 1 1.450 4.000 21.71
5 1 1.147 3.13 38.74
10 1 0.931 2.518 50.71

Efficiency Vs Dosage
60

50

40
Efficiency %

30

20

10

0
0 2 4 6 8 10 12
Dosage g/l

FIG 4.12 Efficiency Vs Dosage of Banana peels

49
 TABLE 4.14

Variation of Time of Contact of Banana peels

Amount of Time Absorbance Chromium Removal efficiency

adsorbent (g/l) duration concentration %
(hour) (ppm)
10 0.5 1.183 3.238 36.62
10 1 0.935 2.53 50.48
10 3 0.877 2.364 53.73

Efficiency Vs Time of contact

60

50

40
Efficiency %

30

20

10

0
0 0.5 1 1.5 2 2.5 3 3.5
Time of contact hr

Fig 4.13 Efficiency Vs Time of contact of Banana peels

50
 CHAPTER 5

SUMMARY & CONCLUSION

5.1 SUMMARY

A comparative graph showing the Chromium removal efficiency of different

biosorbents at varying dosage and varying time is plotted.

COMPARISON FOR VARIATION OF DOSAGE WITH

EFFICIENCY FOR DIFFERENT BIOSORBENTS
70

60

50

40

30

20

10

0
2.5 g/l 5 g/l 10 g/l

FISH SCALES RICE HUSK RICE HUSK ASH

CROWN FLOWER BANANA STALK BANANA PEELS

FIG 5.1 Comparison of Variation of Dosage with Efficiency for different biosorbents

51
 COMPARISON FOR VARIATION OF TIME OF CONTACT
WITH EFFICIENCY FOR DIFFERENT BIOSORBENTS
80

70

60

50

40

30

20

10

0
30 MINS 1 HOUR 3 HOUR

FISH SCALES RICE HUSK RICE HUSK ASH

CROWN FLOWER BANANA STALK BANANA PEELS

FIG 5.2 Comparison of Variation of Time of Contact with Efficiency for different
biosorbents

5.2 CONCLUSION

Of all the biosorbents used in this project, Fish scales and Rice Husk ash provided
higher efficiency than all other biosorbents. The best biosorbent for duration is fish
scales which provided 67.88% of Hexavalent Chromium removal for a dosage of 10 g/l
in 3 hour. The best biosorbent for smaller amount of adsorbent is Rice Husk ash which
provided 64.53% efficiency for a dosage of 10 g/l in 1 hour.

5.3 SCOPE OF THE PROJECT

Further studies on this project can be conducted by combining different

biosorbents of higher efficiency in removing different heavy metal ion to make a
combination which can remove all the heavy metals present in the industrial effluent at
a comparatively lower cost.

52
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