Republic of the Philippines

DEPARTMENT OF EDUCATION

Region XI

Atty. Orlando S. Rimando National High School

Binuangan, Maco Compostela Valley

MOSSES AS A REMOVER OF LEAD (Pb) FROM AQUEOUS SOLUTION

A Science Investigatory Project

Presented to the Science Department of

Atty. Orlando S. Rimando National High School

In partial fulfilment of the requirements in Independent Research II

Nur-aine L. Hajijul

Chariz R. Moldez

Jennis Gail T. Siaton

Regine M. Jamin

March 2019

1
 Table of Contents

Chapter 1

Tittle Page ....................................................................................................

Introduction ..................................................................................................

Research Questions .....................................................................................

Hypothesis ...................................................................................................

Review of Related Literature ........................................................................

Scope and Limitations ..................................................................................

Significance of the Study ..............................................................................

Definitions of Terms .....................................................................................

Chapter 2

Methods .......................................................................................................

Materials.......................................................................................................

Testing .........................................................................................................

Locale of the Study.......................................................................................

Research Design ..........................................................................................

Sampling Procedure .....................................................................................

Statistical Treatment.....................................................................................

Chemical Safety Post-Procedure .................................................................

2
 Acknowledgement

CHAPTER 1

3
INTRODUCTION

Lead-contaminated water is a serious environmental concern that has recently

proved to be disastrous when left untreated. Lead rarely occurs naturally in water; it

usually gets into the water from the delivery system. Lead pipes are the main contributor

to high lead levels in tap water (Centers for Disease Control and Prevention, 2015). Lead

poisoning can result from a single high-level (acute) exposure or through a number of

smaller repetitive (chronic) exposures. Lead has no beneficial effect on humans. Once it

has been ingested into the body, lead is distributed in the bloodstream to red blood cells,

soft tissues and bone. In rare instances, damage to the brain and central nervous system

also may occur.

Lead has been identified in at least 1,272 of the 1,684 hazardous waste sites that

have been proposed for inclusion on the EPA National Priorities List (NPL) (HazDat,

2006). However, the number of sites evaluated for lead is not known. Of these sites, 1,258

are located within the United States, 2 are located in Guam, 10 are located in the

Commonwealth of Puerto Rico, and 2 are located in the Virgin Islands. (Fedrip, 2005).

Health effects from lead exposure are a concern both at the workplace and in the

home. Because of this, the student researchers are finding an alternative way to prevent

further effect of lead to streams of water. This study is more revised and its scope is just

4
around our municipality so it is affluent to conduct this study. This study is intended for

people that lives around lead contaminated waters, especially fishermen.

RESEARCH QUESTIONS

This study is to be conducted to answer the following problems and questions:

1. What is the advantage of using the mosses as an alternative source against lead

(Pb)?

2. What components do the mosses have to remove the lead (Pb) from aqueous

solution?

HYPOTHESIS

1. Mosses can’t be used as a remover for lead (Pb).

2. Mosses is not effective in removing lead (Pb) from aqueous solution.

REVIEW OF RELATED LITERATURE

MOSSES

Mosses, one of the major groups of bryophytes, are ubiquitous and dominant

components of ground-layer vegetation in both upland forests and peatlands across the

boreal biorne. These plants have received attention in several recent reviews for their

importance in regulating soil hydroclimate and nutrient cvclinz in boreal ecosvstems (van

Breemen 1995; Turetsky 2003; Nilsson and Wardle 2005).

5
 Recent studies affiliated with the Bonanza Creek Long Term Ecological Research

(BNZ-LTER) program also have documented relationships between moss composition

and ecosystem parameters such as aboveground tree productivity and soil C storage

(Hollingsworth et al 2008) and have suggested that moss abundance plays a critical role

in post-fire successional trajectories (Johnstone et al. 2010) and perrna-frost stability

(Jorgenson et al. 2010).

Generally, moss biomass on the forest floor tends to be effective in buffering soils

from variation in atmospheric climate because of its low thermal conductivity, high

porosity, and high water holding capacity (Rydin and McDonald 1985; O'Donnell et al.

2009). In addition to controlling soil climate, mosses regulate C and nutrient cycling in

boreal ecosystems by (i) contributing to ecosystem net primary productivity, (ii)

contributing to ecosystem N inputs via N-fixing symbioses with cyanobacteria (DeLuca et

al. 2002), (iii) effectively sequestering atmospheric N, thereby preventing N uptake by

vascular plants (Li and Vitt 1997), (iv) producing recalcitrant litter that resists microbial

breakdown (Hobbie et al. 2000; Lang et al. 2009), and (v) regulating plant community

structure by inhibiting seedling germination and the success of many boreal tree species

(Johnstone et al. 2010).

Because mosses possess unique physiological and ecological traits that influence

soil climate, nutrient cycling, and vascular plant germination, changes in moss structure

and function are important for predicting future landscape patterns and processes in a

changing climate across the boreal region. While the BNZ-LTER program has long

recognized the linkages between mosses, peat accumulation, and permafrost dynamics,

6
and the importance of these linkages for boreal forest nutrient cycling (Van Cleve et al.

1991),

Mosses have spread all around the world and are found in wet environments

such as rainforests, wetlands and alpine ecosystems. They are also common in urban

areas with a wet climate and often establish on driveways, sidewalks, brick walls and

other man-made structures. Mosses require water to reproduce which is why they

struggle to survive in drier climates.

Mosses are important for a number of reasons and from many di􀁷erent aspects of

life on Earth. For insects and other invertebrates, mosses can provide a great habitat and

source of food. At a larger scale, mosses perform a number of functions that help

ecosystems perform e􀁷ectively such as filtering and retaining water, stabilizing the

ground and removing CO ‚ from the atmosphere.

Humans have also utilized mosses for a number of reasons. Traditionally, moss

has been used for packing food, helping to insulate houses, and peat formed from semi-

decomposed Sphagnum moss was used as a fuel in the Northern Hemisphere. More

recently, mosses have been used in the florist trade.

The role of mosses in ecosystem succession is likely to be affected by interactions

between directional climate change and climate-mediated disturbances such as wildfire.

Higher nutrient availability in warmer and more oxic soil conditions is expected to favor

vascular plants at the expense of mosses, which tend to be adapted to nutrient-poor

conditions. Increased vascular biomass and litter production can also inhibit moss growth

by shading and (or) burial (Van Wijk et al. 2003). Such shifts in vascular species

7
composition at the expense of mosses below) will influence fuel loading and flammability,

with potential consequences for fire frequency and severity.

Moss cover on the forest floor has a strong influence on the spatial patterns and

overall severity of combustion during boreal wildfires. In general, ground-layer fuels

dominate combustion during many boreal wildfires, with thick layers of live moss and dead

moss tissue serving as an important fuel type (Amiro et al. 2001).

Due to its low bulk density, feather moss biomass can dry out quickly and serve

as flammable fuels. Sphagnum mosses, particularly species that form hummocks such

as S. fuscum, have high water retention due to a dense canopy structure, efficient wicking

ability (Rydin and McDonald 1985), and slow decomposition rates that maintain

macropore structure over time (Turetsky et al. 2008).

Thus, while the mosses in hummocks exist farther from the water table than in

hollows, these microforms tend to have higher surface soil moisture contents. During

periods of drought, high water retention in hummocks can have an important influence on

vascular stress and survival. Sphagnum hummocks also tend to be the last ground-layer

fuels to combust, often escaping deep burning during boreal wildfires (Shetler et al. 2008;

Johnstone 2010). This resistance to burning creates "Sphagnum sheep" (unburned

Sphagnum hummocks interspersed within charred hollows and flat microforms) that play

an important role in post-fire soil C storage (Shetler et al. 2008).

Lead (Pb)

Lead is the most important toxic heavy element in the environment. Due to its

important physico-chemical properties, its use can be retraced to historical times. Globally

8
it is an abundantly distributed, important yet dangerous environmental chemical

(Mahaffay, 1990). Its important properties like softness, malleability, ductility, poor

conductibility and resistance to corrosion seem to make difficult to give up its use. Due to

its non-biodegradable nature and continuous use, its concentration accumulates in the

environment with increasing hazards.

Human exposure to lead and its compounds occurs mostly in lead related

occupations with various sources like leaded gasoline, industrial processes such as

smelting of lead and its combustion, pottery, boat building, lead based painting, lead

containing pipes, battery recycling, grids, arm industry, pigments, printing of books, etc.

Though its widespread use has discontinued in many countries of the world, it is still used

in many industries like car repair, battery manufacturing and recycling, refining, smelting,

etc.

Lead is a highly poisonous metal affecting almost every organ in the body. Of all

the organs, the nervous system is the mostly affected target in lead toxicity, both in

children and adults. The toxicity in children is however of a greater impact than in adults.

This is because their tissues, internal as well as external, are softer than in adults. Long-

term exposure of adults can result in decreased performance in some tests of cognitive

performance that measure functions of the nervous system. Infants and young children

are especially sensitive to even low levels of lead, which may contribute to behavioural

problems, learning deficits and lowered IQ (Rubin & Strayer, 2008).

Long-time exposure to lead has been reported to cause anaemia, along with an

increase in blood pressure, and that mainly in old and middle aged people. Severe

damage to the brain and kidneys, both in adults and children, were found to be linked to

9
exposure to heavy lead levels resulting in death. In pregnant women, high exposure to

lead may cause miscarriage. Chronic lead exposure was found to reduce fertility in males

(Sokol & Berman, 1991). Blood disorders and damage to the nervous system have a high

occurrence in lead toxicity.

Several methods are used to detect elevated blood lead levels. The presence of

changes in blood cells visible under the microscope or deletion of dense lines in the bones

of children seen on X-ray are signs used for detecting lead poisoning. However the main

tool to detect elevated levels of body lead is to measure the level

of lead in blood samples. This test gives however only an account of lead present in

circulating blood but cannot show how much lead is stored in the body. As of 2012, the

Centers for Disease Control and Prevention (USA) have set the standard elevated blood

lead level for adults to be 10 μg/dL and for children 5 μg/dL of the whole blood (CDC,

2012).

All along human history, lead poisoning has been reported to have severe effects.

Occasional lead poisoning was found to be caused by lead salts used in pottery glazes

leached by acidic fruit juices. Beethoven's death has been treated in various reports.

Many of them have concluded that he died because of the toxic doses of lead-based

treatment administered by his doctor. Analysis of his hair was found to contain elevated

levels of lead (Mai, 2006). It is also assumed that in the eighteenth and early nineteenth

century lead was illegally added to wine both as a sweetener and to make it appear fresh

(Mai, 2006). Lead poisoning is believed to be primarily responsible for the collapse of the

Roman Empire, in which lead acetate was used as a sweetener of wine. Its prolonged

use was considered to have caused dementia to many Roman emperors.

10
 Lead poisoning has also been found to be the cause of anaemia in a number of

cases as lead inhibits porphobilinogen synthase and ferrochelatase, preventing both

porphobilinogen formation and the incorporation of iron into protoporphyrin IX, which

prevents heme synthesis (Cohen et al., 1981) or causes ineffective heme synthesis and

subsequently microcytic anaemia. One of the mechanisms by which lead interferes with

cognition is that it acts as calcium analogue which interferes with ion channels. It has

been observed that Pb is a potent reversible and selective blocker of voltage-dependent

calcium channels at low concentrations (Busselberg et al., 1993). In a recent study, the

authors showed that the toxic effects on blood cells of rats caused by lead nitrate were

alleviated by sodium selenite. They also showed that effects of lead nitrate were more

harmful in diabetic than in non-diabetic rats (Bas et al., 2015). Oxidative stress was

studied by low level lead exposure in first grade Uruguayan children, suggesting its

potentially adverse effects on oxidative stress (Roy et al., 2015). Impaired respiratory

function was observed in workers exposed to lead with elevated blood lead concentration

and zinc protoporphyrin concentration (Jurdziak et al., 2015).

Lead is a common environmental pollutant. Exposure to lead occurs mainly at

occupational sites, production of lead-acid batteries or pipes, metal recycling and

foundries (Woolf et al., 2007). Children living near such places are also at risk of elevated

blood lead levels. In August of 2009, 2000 children living near zinc andmanganese

smelters were found to be poisoned with lead, an incident which resulted in riots (Watts,

2009).

Other common things which cause lead exposure are lead in the air, household

dust, soil, water, and commercial products (Rossi, 2008). In cases of chronic exposure,

11
lead often sequesters in the highest concentrations first in the bones then in the kidneys.

According to the US Centres for Disease Control and Prevention and the World Health

Organization, a blood lead level of 10 μg/dL or above is a cause for concern. However

there is no threshold which lead exposure can be considered safe. It has been found to

impair development and have harmful effects even at lower levels (Rossi, 2008; Barbosa

et al., 2005).

A variety of compounds formed by lead exists in the environment in different forms

(Grant, 2009). Poisoning and its features also differ between organic and inorganic lead

(Kosnett, 2007). Organic lead poisoning is now very rare around the world because of

withdrawal of organic lead compounds as gasoline additives. Nevertheless, such

compounds are still used in industrial settings. Organic lead compounds cross the skin

and respiratory tract easily and quickly, affecting predominantly the central nervous

system.

Bioremediation

Bioremediation can be defined as any process that uses microorganism isms or

their enzymes to return the environment altered by contaminants to its original condition.

Bioremediation may be employed in order to attack specific contaminants, such as

chlorinated pesticides that are degraded by bacteria, or a more general approach may be

taken, such as oil spills that are broken down using multiple techniques including the

addition of bio surfactant to facilitate decomposition of crude oil by bacteria (Juwarkar et

al. 2008).

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 Bioremediation may be either aerobic (Wiegel and Wu 2000; Bedard and May

1996) or anaerobic (Komancova et al. 2003). Due to the problem associated with either

of this method to treat highly complex compounds, sometime sequential anaerobic–

aerobic bioremediation processes are also adopted to remedi ate contaminated sites

(Master et al. 2002). Remediation using fungal strains in some cases proved as a highly

effective remediation approach (Kuba´tova´ et al. 2001).

Bioremediation is an increasingly popular alternative to conventional chemical

methods for treating waste compounds and media with the possibility to degrade

contaminants, since it uses natural microbial activity mediated by different consortia of

microbial strains. Many studies on bioremediation have been reported and the (A.A.

Juwarkar et al.) scientific literature has revealed the progressive emergence of various

advances in bioremediation techniques. This chapter emphasizes on recent trends in

bioremediation techniques/processes for decontamination of different environmental

matrices.

SCOPE AND LIMITATIONS

This experiment is conducted to investigate the effectiveness of horticultural

mosses in the removal of lead from aqueous solution. We think that mosses can function

as an excellent lead adsorbent in the protonema stage of development. This valuable

ability means that moss protonema will likely make exceptional wastewater cleaners in

mining and chemical industries. The mosses will be collected in Maco Compostela Valley.

13
We will review the use of moss in the removal of heavy metals from wastewater and will

conduct laboratory studies to evaluate the adsorption capacity of mosses for lead.

SIGNIFICANCE OF THE STUDY

The findings of the study will redound to the benefit of society considering that it

can enable us to prevent harmful effects on our central nervous system and reproductive

system caused by contaminated wastewater. We can also prevent continuous toxic level

of exposure and consequently clinical poisoning caused by excessive contamination of

lead in industrial activities that is generally contained in the air, food and water we take

in. This study can also inspire fellow researchers especially to those who will be

conducting this kind of research, to expand more on the current study and to be well

informed about its benefits to use it in a better way.

DEFINITION OF TERMS

Mosses. Mosses are a phylum of non-vascular plants. They produce spores for

reproduction instead of seeds and don’t grow flowers, wood or true roots. Instead of roots,

all species of moss have rhizoids. The mosses sit within a division of plants called

the Bryophyta under the sub-division Musci. (Basic Biology, 2018). This is the main

variable of the study.

14
 Lead (Pb). A chemical element with a symbol Pb and atomic number 82. A heavy

metal that is denser than the most common materials (Thoughtco.com). This serves as

our dependent variable of the study.

Industrial Wastes. waste produced by industrial activity which includes any

material that is rendered useless during a manufacturing process such as that of factories,

industries, mills and mining operations. (Encyclopedia, 2018)

Aqueous Solution. A solution in which the solvent is water. It is mostly shown in

chemical equations by appending to the relevant chemical formula. (Basic Biology, 2015).

Adsorption. The adhesion of atoms, ions, or molecules from a gas, liquid or

dissolved solid to a surface. This process fills of the adsorbate on the surface of

adsorbent. Thus process differs from adsorption, in which a fluid is dissolved by or

permeates a liquid or solid, respectively. (Encyclopedia, 2014)

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 CHAPTER 2

METHODS

This chapter presents all the methods and instruments employed in the procedure

and the statistical treatments utilized in analyzing the data gathered in order to determine

the Effectiveness of Funaria Hygrometrica to test its capability in adsorbing lead from

contaminated wastewater.

RESEARCH DESIGN

The researcher used pretest-posttest true experimental design. The experiment

will be carry out using mosses. The materials are dried for 4 hours at 90 ± 2°C and then

grounded and sieved to particles with size of 1-2 mm. The mosses will be collected from

Anislagan, Maco, Compostela Valley Province.

MATERIALS
500 grams of dried mosses
100 mL of lead
500 mL of distilled water
Safety gloves
Face mask
Clean container
5 clean jars
Laboratory apparatus

16
TESTING

LOCALE OF THE STUDY

This study will be conducted at Prk 1 Anislagan Maco. We still seek the approval

from the person who is in charged in managing water supplies located at Prk 1 Anislagan

Maco to ensure that our collection of contaminated wastewater is not illegal. The

collection of Mosses was collected at Prk.1 Anislagan Maco. Mosses is prevalently

available not only within Maco but even in the surrounding communities.

Map of Maco Prk. 1 Anislagan, Maco, Compostela Valley Province

PRE-PROCEDURE

Research Proposal will be submitted to the (Place sa collection sa inyong

contaminated soil) to secure approval for the use of their contaminated wastewater.

Further, another copy of the research proposal will be submitted to Engr. Paolo Marrence

D. Tuyor, from Tribal Mining Corporation, Tiboli, South Cotabato, requesting his expertise

17
in mining engineering as the study qualified scientist and provide the researcher technical

guidance and close supervision for the entire duration of the experimentation. Approval

for the use of Mosses will be collected in Prk. 1 Anislagan Maco.

SAMPLING PROCEDURE

Preparation of adsorbent

The adsorption experiments are performed by batch technique, mixing samples

of cca. 0.125 g of mosses with volumes of 25mL solution of known concentration of lead

(34-507 mg Pb (ll)/L), in a 10 mL conical flas. The initial ph of work solutions is obtained

adding 2.5 mL of 2N HNOӡ solution (for pH ■ 4.0 and 6.0), and was measured with a

radelkis OK-281 pH/ion-meter, equipped with a combined glass electrode. After a

determined time (usually 24 hours) with intermittent stirring, the phases are separated by

filtration and the Pb (II) concentration in filtrate is analyzed spectrophotometrical with 4-(

2-pyridylazo)- resorcinol (Digital Spectrophotometer S 104 Dӡ ■ 530 nm, 1 cm glass cell

against a blank solution (13), using a prepared calibration graph.

Effect of Mosses Pb adsorption

In the experiment concerning the effect of mosses, a range of sorbent samples

from 0.125 to 1.000g were used. For the contact time experiment, we use the same

procedure as describe above, with the differences that the samples were filtrated after a

specific period of time.

18
 The residual lead concentration (C Pb,mg/g) and the percent of lead removal (R,%)

were calculated according to their definition. The selectivity coefficients are calculated as

the ration between the lead concentration and interfering ions concentration which gives

a 5% change of removal percent in a reference solution.

Effect of agitation on Pb adsorption

The agitation on the shaker was decided to be 100 rpm and 165 rpm to make the

process more efficient for a contact time of 60 mins and 90 mins after the pH adjustment

and addition of .1 g of adsorbent was done. Afterwards filtration using Whatmann filter

paper number 1 was done followed by giving the samples to Atomic absorption

Spectroscopy to check the adsorption.

Effect of contact time on Pb adsorption

The contact time used was 60 mins and 90 mins during the agitation period after

which again filtration occurred and adsoption was checked by subjecting the samples to

the Atomic absorption Spectroscopy.

19
Chemical Safety Post-Procedure.

The contaminated biomass by-products of phytoremediation technique need

volume reduction through various techniques as follows in order to handle it safely.

1 All types of 27 protective measures were used while doing the experiments.

Safety measures included wearing safety gloves, masks and goggles and lab

apron. All the equipment used in this study were cleaned properly to prevent

cross contamination.

2 The glassware was cleaned using a standard procedure, which led washing of

the item with tap water and soap followed by a nitric acid (50% by volume) wash

and finally distilled water wash.

3 Proper dilutions of filtered sample were prepared and analyzed by Atomic

Absorption (AA) Spectrometry (EPA, 1996). Digestion of samples were done in

triplicate and performed under a hood to ensure safety.

4 All volatile materials must be handled in a hood with the exhaust fan on. When

refluxing any solutions, perform this operation under a hood, and use boiling

beads in the reflux vessel to prevent any excessive "bumping" and possible

glass breakage.

5 Disposal of all used solutions in an approved manner as directed by the

qualified scientist.

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STATISTICAL TREATMENT

The following statistical tool aided the researchers in providing answers to the

research questions posted in the study.

Mean. This statistical tool will be used to answer research question number 2

(What is the advantage of using the mosses as an alternative source against lead?)

21
 Chapter 4

DISCUSSION

The primary purpose of this study is to use mosses as a remover of lead from any

aqueous solution that can be a great help for any water sources that are contaminated

with lead. Earlier studies regarding mosses used filtering which can be expensive and

22
hard to find in some places. This study utilized the abundance of mosses in the

researcher’s locality by using dried mosses as a remover of lead.

CONCLUSION

With the support of the aqueous solution result that proved the removability of our

product, makes it suitable for mass production and substitution of the commercial filtering

in their everyday use. However further research and experimentation should be done to

increase the efficiency of the product.

RECOMMENDATIONS:

 Find a way to completely remove the lead from aqueous solution.

 Try to make the mosses more durable for multi-purposes.

 Add more tests to guarantee the quality of the mosses.

REFERENCES

MATERIALS AND METHODS

23
Lead(II) solutions

All chemicals were analytical grade and purchased from Merck (Darmstadt, Germany). The lead

stock solution was prepared by dissolving an accurate quantity of PbCl2 in deionized water. Other

lead working solutions were freshly prepared by diluting the lead stock solution with deionized

water. The solution pH was adjusted by adding 0.1 M H2SO4 or 0.1 M NaOH, as required.

The biosorbent

This type of aquatic plants was chosen because their high capacity for assimilating nutrients, toxic

organics and heavy metals, which has been related in monitoring programs of rivers water quality.

Another’s relevant reasons: this material is abundant in nature, require little processing and there
is

a bit applications in literature about them.

The aquatic moss Fontinalis antipyretica was taken from the Selho River, a tributary of the Ave

River in Northern Portugal. The background metal content in mosses was considered to be of

-1
natural origin and negligible (sensibly 114 μg g ). The samples were rinsed with river water in situ

and then in the laboratory with deionized water, selecting only the green parts of the plants. The

plants were dried in an oven at 70ºC for 24 h and then ground in a RETSCH ZM 100 ultra-

centrifugal mill at 1400 rpm for approximately 90 seconds. The fraction with particle size between

150 μm - 300 μm was selected for this study.

Equilibrium experiments

Equal amounts of 100.0 (±0.1) mg of the biosorbent were accurately weighted and transferred to

24
 -1
100 ml glass bottles and 50 mL of lead solution with concentration between 10 and 100 mg L
was

added to each bottle. The bottles were then shaken for 24 h at constant room temperature
(20±1ºC)

using a rotary shaker (P-Selecta Rotabit). Each experiment was performed in duplicate and blank

solutions (metal solution without biosorbent) were shaken simultaneously. The contact time was

previously determined from kinetic studies, carried out at the same temperature and stirring rate,
as

that required to attain equilibrium. After equilibrium was reached the liquid phase was separated

from the biosorbent by vacuum filtration through 0.45 μm Gelman Sciences sterilized membranes,

and the supernatant was analysed for the residual lead. The metal uptake was calculated by the

equation

−=
)//()( VmCCq
0 adsee
( 1 )

where V (L) is the volume of lead solution,

0

-1
C (mg L ) is the initial lead concentration,
e

-
C (mg L

1
ads
) is the concentration of lead in solution at equilibrium, and

m (g) is the mass of biosorbent (dry

weight).

Kinetic experiments

The influence of the initial metal concentration and biosorbent dosage on biosorption rate was

25
studied in batch mode.

Erlenmeyer flasks with 100 ml capacity were used in this study, each one containing 50 ml of metal

solution of a known initial concentration and a pre-weighted amount of dry biomass. The

-1
suspensions were mildly stirred for 300 min on a rotary shaking machine at 140 U min .

Biosorption kinetic experiments were performed at constant temperature of 20ºC, metal

-1 -1
concentrations of 10 and 100 mg l and biosorbent dosage of 2 g L . In all cases, the initial pH
was

adjusted in the range 5.0-5.2 by adding 0.1 M H2SO4 or 0.1 M NaOH, as required. Samples were

taken from each flask at pre-determined time intervals, starting at 3 min and ending at 300 min.

Then, the solution was filtered (0.45 μm Gelman Sciences sterilized membranes) and the metal

concentration in aqueous solution was measured. All the experiments were conducted in duplicate.

26