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International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056
Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072
© 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7144
An Investigation into the efficacy of Fungal Biomass as a Low Cost Bio-
adsorbent for the removal of Lead from aqueous solutions
Swati Rastogi1, Jeetendra Kumar2, Rajesh Kumar3
1Research Scholar, Rhizosphere Biology Laboratory, Department of Microbiology, Babasaheb Bhimrao Ambedkar
University (a central university) vidya vihar raebareli road, Lucknow, India
2M.Sc. Student, Rhizosphere Biology Laboratory, Department of Microbiology, Babasaheb Bhimrao Ambedkar
University (a central university) vidya vihar raebareli road, Lucknow, India
3Professor, Rhizosphere Biology Laboratory, Department of Microbiology, Babasaheb Bhimrao Ambedkar
University (a central university) vidya vihar raebareli road, Lucknow, India
---------------------------------------------------------------------***---------------------------------------------------------------------
Abstract - Some of the heavy metals such as lead (Pb2+)
even in low concentration pose a threat to human wellbeing
and other life forms. Various anthropogenic and industrial
sources discharge this toxic metal into the biosphere. The
present study explores the efficiency of untreated dead
biomass of Penicillium sp. (a fungus)inthebio-sorptionof Pb2+
ions from aqueous solutions. Different factors viz., initial Pb2+
ion concentration, adsorbent dose and contact time were
studied. The maximum adsorption percentage (78.03%) of
Pb2+ was found under the optimum conditions of 10 mg/l of
Pb2+, an adsorbent dose of 1g/L and contact time of 2 hours.
Langmuir adsorption isotherm was best fitted for the present
study (R2=0.9984). Bio-sorption reaction mechanism was
explained through FTIR analysis of fungal biomass which
revealed the presence of carbonyl, methylene, phosphate,
carbonate and phenolic groups and theirpossibleinvolvement
in the Pb2+ ions bio-sorption process. SEM and EDX details
provide the structuralcharacterizationandopticalabsorption
peaks of dead fungal biomass and explain its surface
morphology in the adsorption and removal of Pb2+ ions from
aqueous solutions respectively. It can be concluded that
untreated dead biomass of Penicillium sp. is a promising,
efficient, low-cost bio-adsorbent for the removal of Pb2+ ions
from the environment and wastewater effluents.
Key Words: Biosorption; low-cost bio-adsorbent;
SEM; FTIR; Langmuir isotherm model
1. INTRODUCTION
Enhanced industrial andeconomic developmenthasresulted
in heavy metal pollution and their vast distribution in the
environment which in turn poses a major challenge for their
effective treatment and management due to their toxic and
persistent nature and accumulation in the food chain [1].
Many conventional technologies including filtration, ion-
exchange, precipitation, coagulation, membrane separation,
solvent extraction are available for mitigation of these
dangerous pollutants but demands high energy and
operation costs even lead to the generation of secondary
pollutants/sludge. Due to which focus has been shifted
towards the application of cost-effective, eco-friendly non-
conventional methodsforremediationoftheseheavymetals.
Bioadsorption is one of the emerging process/methods
which is considered to be beneficial for the removal of metal
ions from aqueous solutions. Microbial biomasseitherliving
or dead as bioadsorbents has been found to be effective in
the amelioration of heavy metal ions from wastewater/soil.
Literature survey supports the use of fungi, algae, and
bacteria as adsorbents for several heavy metals [2] [3].
Recently, lead removal from soil and aqueous samples have
been reported by many fungal species like Aspergillus niger
[4][6][11][14], Trichoderma reesi [4], Mucor arcindloides [4],
Saccharomyces cerevisiae [4][9] , Penicillium austurianum [4],
Penicillium verrucosum[5], Botrytiscinerea[7], Phanerochaete
chrysosporium [8], Mucor rouxii [10], Aspergillus fumigatus [11],
Penicillum simplicissimum [11], Trichoderma
asperellum[11],Penicillium chrysogenum[12] [13] [15], Aspegillus
nidulans [15], Aspergillus flavus [15], Rhizopus arrhizus [15],
Trichoderma viride [15].
The utility of fungal biomass as bioadsorbent is in the fact as
are easy to separate, rarely sensitive to nutrient variation,
pH etc, has less nucleic content in biomass, easily cultivated
for large scale production, provides large surface area for
adsorption reactions and are non-toxic [16][17].
International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056
Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072
© 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7145
The objective of the present study was to explore the lead
removal potential of fungal biomass as biosorbent from
aqueous solution in the batch process. The effect of
parametersinfluencing biosorptionprocessesssuchasinitial
lead concentration, adsorbent dose and contact time were
also studied. The affirmation of lead biosorption on fungal
biomass was done through Fourier transform infrared
spectroscopy (FTIR) and scanning electron microscopy
combined with X-ray energy dispersive spectrometer(SEM-
EDX) analysis upon exposure to lead ions in the aqueous
solutions.
2. MATERIAL AND METHODS
2.1 Preparation of stock and standard
solutions of lead nitrate [Pb (NO3)2]
In a clean beaker containing 100ml distilled water, 1.59g of
Pb(NO3)2 was dissolved with constant stirring to get a
10000ppm stock solution and afterward 10µL, 20µL and
30µL were pipetted out from ittomakestandardsolutions of
5ppm, 10ppm and 15ppm in 20ml distilled water
respectively. The standard solutions were prepared fresh
and pH was measured each time before the experiment was
conducted.
2.2 Preparation and characterization of bio-
adsorbent
Dead and dried fungal biomassof Penicilliumsp. wasusedfor
removal of Pb2+ ions from aqueous solutions. It was
procured from microbiology department of Lucknow
University in dried and dead form only. Thecharacterization
of fungal adsorbent was done with the help of technique
scanning electron microscopy-energy dispersive X-Ray
(SEM-EDX) and Fourier-transform infrared spectroscopy
(FTIR).
2.2.1 Fourier transformed infrared (FTIR)
spectral analysis
For FTIR studies, 4-5 mg of biosorbent (the lead-treatedand
the untreated fungal biomass)wascoveredin400mgof KBr.
Round discs were obtained by pressing the ground material
with the aid of a bench press and further readings were
measured on NicoletTM 6700 model (ThermoScientific,USA)
FTIR spectrometer.
2.2.2 Scanning electron microscopy combined
with X-ray energy dispersive spectrometer
(SEM-EDX) analysis
The surface morphology and lead binding pattern in the
lead-treated and the untreatedfungal biomasssampleswere
recorded usingscanningelectron microscopycombinedwith
X-ray energy dispersive spectrometer (SEM-EDX, JEOL, JSM
6490 LV, Japan).
2.3 Batch adsorption experiments
Batch equilibrium experiments were performed with 20ml
of a standard solution of concentration 5mg/l, 10mg/l and
15mg/l and a desired adsorbent dose of 0.1g to 1g. The
parameters studied include initial metal ion concentration,
contact time and adsorbent dose. The mixture was
transferred to 50ml Erlenmeyer flasks and stirred for 2h to
6h, filtered using Whatman filter No. 42 and analyzed for
residual metal concentration through inductively coupled
plasma- mass spectrometry (ICP-MS). All the experiments
were performed at room temperature conditions.
The amount of lead adsorbed onto fungal biomass was
calculated by using the following expression Eq. (1)
(1)
Where qe is the quantity of lead biosorbed onto fungal
biomass as mg/g; V is the volume of lead solution inliters; Co
and Cf are initial and final concentrations (mg/l) of lead in
the solution and M is the weight of biomass taken for the
batch experiment in g.
2.4 Adsorption isotherm Models
The mathematical relationshipbetweentheconcentrationof
adsorbate before and remained after the sorption
experiment is explained by using adsorption isotherm
models. Langmuir and Freundlich adsorption isotherm
models are used in this study. These are commonly used to
assess the sorption capacity of bio-adsorbent in the
adsorption system.
2.4.1 Langmuir Isotherm
It is generally based on monolayer adsorption process i.e.,
once all the sites available on adsorbent are filled, no more
adsorption process can take place or say it reaches the
saturation point.
The general and the linearized form of this isotherm are
represented in equations (2) and (3) respectivelyasfollows:
(2)
(3)
Where qe is the amount of sorbed metal at equilibrium
(mg/g), qmax is the monolayer sorption capacity (mg/g), b is
the Langmuir constant, Ce is concentration of metal ions in
solution at equilibrium.
International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056
Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072
© 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7146
When a plot of (Ce/qe) versus Ce is plotted, a straight line
with a slope of (1/qmax) and an intercept of (1/bqmax) must
be obtained.
2.4.2 Freundlich isotherm
Heterogenous surfaces involving adsorption processes are
explained by using Freundlich isotherm. It is empirical and
does not assume monolayer adsorption but is quiteuseful to
explain adsorption data andfractionofactivessitesavailable
for sorption study.
The general and the linearized form of this isotherm are
represented in equations (4) and (5) respectivelyasfollows:
(4)
(5)
Where Ce is the equilibrium concentration (mg/g), qe is the
adsorbed amount at equilibrium (mg/g),Kf istheadsorption
capacity (L/mg) and 1/� is adsorption intensity, it also
indicates the relative energy distribution and the
heterogeneity of the adsorbate sites.
3. RESULTS AND DISCUSSION
3.1 Effect of initial metal ion concentration
The effect of initial metal ion concentration was determined
at operational conditions of 0.1g adsorbentdoseandcontact
time of 2h at pH 5.5 for initial metal ionconcentrationfrom5
mg/l to 15 mg/l. Sorption capacity (qe) was found to be
increased from 0.42 mg/g to 1.136 mg/g indicating an
increase in adsorption rate with increase in initial metal ion
concentration (Fig. 1) and reduced amount of metal ions in
the aqueous solution. These findings could be explained as
there is an acting huge force for rapid mass transfer, more
the concentration of initial metal ions more will be
adsorption rate.[18][19]
Fig.1. Initial metal ion concentration Vs % Adsorption
3.2 Effect of contact time
An investigation into the adsorption process in relation to
contact time is an important parameter as it can help us to
assess the efficiency of adsorption and to determine the
saturation point with constant adsorbent dose and fixed
initial metal concentration. Because at the beginning of the
sorption process, more of the actives sites on the adsorbent
is available and unoccupied and with the increase in time
actives sites begins to get saturated by the metal ions
present in the solution thereby decreasing the adsorption
rate. [20] In the present study, the effect of contact time was
determined at 10ppm/20ml with 0.1g adsorbent dose at pH
5.5 with contact time range from 2h to 6h and results
showed decrease in concentration of metal ions in the
solution and increased adsorption rate as up to 6 hours
which can be explained due to unsaturation of actives sites
and were still available for adsorption process (Fig. 2).
Fig.2. Contact time Vs % Adsorption
3.3 Effect of adsorbent dose
There is a direct relationship between adsorbent dose and
percent adsorption. As we increase the dose of adsorbent
and keep the initial metal ion concentration constant, more
active sites are available for the metal ion to get adsorbed at
the surface of adsorbent [21]. In the present study, the effect
of adsorbent dose was determined at fixed 10ppm/20ml
initial metal ion concentration and a contacttime of6hour at
pH 5.5 and adsorbent dose ranges from 0.1g to 1g and
results indicates increased adsorption rate as more actives
sites are available with increased adsorbent dose at fixed
concentration of metal ion (Fig. 3).
Fig. 3 Adsorbent dose Vs % Adsorption
International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056
Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072
© 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7147
3.4 SEM-EDX and FTIR analysis
SEM-EDX analysis tells us about the surface morphologyand
structural characterization of adsorbent along with their
chemical composition and relative spectral energies.Results
of lead untreated and treated adsorbentareshowninfig.4,5
and 6 respectively. An attempt was made to explore the
surface of dry fungal adsorbent for any modification which
could have been occurred due to adsorption of lead ions and
fig. 4(b) indicates rough irregular morphology containing
dead ruptured hyphae of fungal biomass loaded with lead
ions in comparison to fig. 4(a) which is taken before
adsorption of lead ions. EDX analysis as shown in fig. 5(
before adsorption) and fig. 6( after adsorption) respectively
revealed change in inorganic chemical constituents by
weight percent and some reduction indicating ion-exchange
could be considered as one of the dominant adsorption
mechanism in lead ion biosorption onto fungal biomass.
[22][23]
(a) (b)
Fig. 4 SEM micrographs of fungal adsorbent before
sorption (a) and after sorption (b) of Pb2+ ions (15ppm)
at 1000X magnification
Fig. 5 EDX spectrum of Fungal adsorbent before
adsorption
Fig. 6 EDX spectrum of Fungal adsorbent after
adsorption
FTIR analysis is done to determine the types of functional
groups present on the surface of the adsorbent. Results ofIR
spectra before and after adsorption are presentedinfig.7(a)
and 7(b) respectively. The largest peak of wave number
3384 cm-1 shown in Figure 7, corresponds to either free or
H-bonded O-H groups that could be present in carboxylic
acids on the surface of the fungal adsorbent. The peak at
2924.2 cm-1 wavenumbercorrespondstotheC-Hstretching
that could exist in the alkanes. As for peaks ofwavenumbers
in the range of 1650.2 to 1314.0 cm-1, are an indication of
the olefinic and aromatic bond, and bands of wavenumber
1235.5 to 573.3 cm-1 are an indication of C-O bond. C-O
bond may be due to carboxylic acids,hydroxygroupsorfiber
carbonaceous that are present in the structure of fungal
adsorbent that is composed of lignin and cellulose. Those
groups probably act as proton donors that once get
deprotonated, the hydroxyl group or the carboxyl group
adsorb the heavy metal ions.
As shown in Fig. 7(b), the peak representing the O-H bond
was only shifted from 3384 cm-1 to 3392.3cm-1,whichisan
insignificant change. Whereas the peak of the C=O bond was
shifted from 573.3 to 607.0 cm-1, and the peak of the C-O
bond was shifted from 1066.8 to 1107.8 cm-1. Those shifts
can be attributed to the surface complexation created
between Pb2+ and the carboxylic acids functional groups;
hence, it is an indication that chemical complexation
between Pb2+ and the carboxyl groups could be one of the
mechanisms partially responsible for the biosorption of
Pb2+ via fungal adsorbent. [24]
(a)
International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056
Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072
© 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7148
(b)
Fig. 7 FTIR spectrum of fungal adsorbent before
adsorption (a) and after adsorption (b)
3.5 Adsorption isotherms
The values of Langmuir and Freundlich constants for lead
ions uptake on dry fungal biomass are given in Table 1 and 2
respectively. The valuesofLangmuirconstantsqmaxand b are
calculated by plotting against 1/qe and 1/Ce and Freundlich
constants KF and n were obtained by the plot of log qe vs log
Ce. The R2 value of Langmuir constant was 0.9984 wasfound
to be higher than R2 value of Freundlich (0.9809), indicating
that Langmuir data fitted well than Freundlich data in
adsorption process (Figure 4 & 5).Itisconsideredto be well-
fitted model if R2 value is close to unity. [25]
Figure 4: Langmuir isotherm
Table 1: Parameters of Langmuir isotherm
qmax (mg/g) b(l/mg) R2
0.16 25.94 0.9984
Figure 5: Freundlich isotherm
Table 2: Sorption Parameters of Freundlich isotherm
Kf n R2
3.80 4.59 0.9809
4. CONCLUSIONS
The present research was designed to investigate the
efficiency of a dry fungal adsorbent for the removal of lead
ions from their aqueous solutions. The effect of parameters
such as initial metal ion concentration, contact time and the
adsorbent dose was selected for adsorption study. The
selected dead biomass of fungus showed a notable metal
adsorption capacity over a limited range of parameters. The
Langmuir adsorption isotherm was found to be best fitted
with the present study.
The dead fungal biomass may be employed in the future for
heavy metal remediation from real wastewater and
comparative study can be assessed between dead and living
fungal biomass. Further, column studies could be done to
compare metal uptake and sorption capacity with different
metal ion concentrations, adsorbent dose and contact time.
Column bed height could be varied to compare adsorption
efficiency.
5. ACKNOWLEDGMENT
Authors are thankful to the Head and Coordinator,
Department of Microbiology, University of Lucknow,
Lucknow for biomass availability and support.
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International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056
Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072
© 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7149
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IRJET- An Investigation Into the Efficacy of Fungal Biomass as a Low Cost Bio-Adsorbent for the Removal of Lead from Aqueous Solutions

  • 1. International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056 Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072 © 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7144 An Investigation into the efficacy of Fungal Biomass as a Low Cost Bio- adsorbent for the removal of Lead from aqueous solutions Swati Rastogi1, Jeetendra Kumar2, Rajesh Kumar3 1Research Scholar, Rhizosphere Biology Laboratory, Department of Microbiology, Babasaheb Bhimrao Ambedkar University (a central university) vidya vihar raebareli road, Lucknow, India 2M.Sc. Student, Rhizosphere Biology Laboratory, Department of Microbiology, Babasaheb Bhimrao Ambedkar University (a central university) vidya vihar raebareli road, Lucknow, India 3Professor, Rhizosphere Biology Laboratory, Department of Microbiology, Babasaheb Bhimrao Ambedkar University (a central university) vidya vihar raebareli road, Lucknow, India ---------------------------------------------------------------------***--------------------------------------------------------------------- Abstract - Some of the heavy metals such as lead (Pb2+) even in low concentration pose a threat to human wellbeing and other life forms. Various anthropogenic and industrial sources discharge this toxic metal into the biosphere. The present study explores the efficiency of untreated dead biomass of Penicillium sp. (a fungus)inthebio-sorptionof Pb2+ ions from aqueous solutions. Different factors viz., initial Pb2+ ion concentration, adsorbent dose and contact time were studied. The maximum adsorption percentage (78.03%) of Pb2+ was found under the optimum conditions of 10 mg/l of Pb2+, an adsorbent dose of 1g/L and contact time of 2 hours. Langmuir adsorption isotherm was best fitted for the present study (R2=0.9984). Bio-sorption reaction mechanism was explained through FTIR analysis of fungal biomass which revealed the presence of carbonyl, methylene, phosphate, carbonate and phenolic groups and theirpossibleinvolvement in the Pb2+ ions bio-sorption process. SEM and EDX details provide the structuralcharacterizationandopticalabsorption peaks of dead fungal biomass and explain its surface morphology in the adsorption and removal of Pb2+ ions from aqueous solutions respectively. It can be concluded that untreated dead biomass of Penicillium sp. is a promising, efficient, low-cost bio-adsorbent for the removal of Pb2+ ions from the environment and wastewater effluents. Key Words: Biosorption; low-cost bio-adsorbent; SEM; FTIR; Langmuir isotherm model 1. INTRODUCTION Enhanced industrial andeconomic developmenthasresulted in heavy metal pollution and their vast distribution in the environment which in turn poses a major challenge for their effective treatment and management due to their toxic and persistent nature and accumulation in the food chain [1]. Many conventional technologies including filtration, ion- exchange, precipitation, coagulation, membrane separation, solvent extraction are available for mitigation of these dangerous pollutants but demands high energy and operation costs even lead to the generation of secondary pollutants/sludge. Due to which focus has been shifted towards the application of cost-effective, eco-friendly non- conventional methodsforremediationoftheseheavymetals. Bioadsorption is one of the emerging process/methods which is considered to be beneficial for the removal of metal ions from aqueous solutions. Microbial biomasseitherliving or dead as bioadsorbents has been found to be effective in the amelioration of heavy metal ions from wastewater/soil. Literature survey supports the use of fungi, algae, and bacteria as adsorbents for several heavy metals [2] [3]. Recently, lead removal from soil and aqueous samples have been reported by many fungal species like Aspergillus niger [4][6][11][14], Trichoderma reesi [4], Mucor arcindloides [4], Saccharomyces cerevisiae [4][9] , Penicillium austurianum [4], Penicillium verrucosum[5], Botrytiscinerea[7], Phanerochaete chrysosporium [8], Mucor rouxii [10], Aspergillus fumigatus [11], Penicillum simplicissimum [11], Trichoderma asperellum[11],Penicillium chrysogenum[12] [13] [15], Aspegillus nidulans [15], Aspergillus flavus [15], Rhizopus arrhizus [15], Trichoderma viride [15]. The utility of fungal biomass as bioadsorbent is in the fact as are easy to separate, rarely sensitive to nutrient variation, pH etc, has less nucleic content in biomass, easily cultivated for large scale production, provides large surface area for adsorption reactions and are non-toxic [16][17].
  • 2. International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056 Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072 © 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7145 The objective of the present study was to explore the lead removal potential of fungal biomass as biosorbent from aqueous solution in the batch process. The effect of parametersinfluencing biosorptionprocessesssuchasinitial lead concentration, adsorbent dose and contact time were also studied. The affirmation of lead biosorption on fungal biomass was done through Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy combined with X-ray energy dispersive spectrometer(SEM- EDX) analysis upon exposure to lead ions in the aqueous solutions. 2. MATERIAL AND METHODS 2.1 Preparation of stock and standard solutions of lead nitrate [Pb (NO3)2] In a clean beaker containing 100ml distilled water, 1.59g of Pb(NO3)2 was dissolved with constant stirring to get a 10000ppm stock solution and afterward 10µL, 20µL and 30µL were pipetted out from ittomakestandardsolutions of 5ppm, 10ppm and 15ppm in 20ml distilled water respectively. The standard solutions were prepared fresh and pH was measured each time before the experiment was conducted. 2.2 Preparation and characterization of bio- adsorbent Dead and dried fungal biomassof Penicilliumsp. wasusedfor removal of Pb2+ ions from aqueous solutions. It was procured from microbiology department of Lucknow University in dried and dead form only. Thecharacterization of fungal adsorbent was done with the help of technique scanning electron microscopy-energy dispersive X-Ray (SEM-EDX) and Fourier-transform infrared spectroscopy (FTIR). 2.2.1 Fourier transformed infrared (FTIR) spectral analysis For FTIR studies, 4-5 mg of biosorbent (the lead-treatedand the untreated fungal biomass)wascoveredin400mgof KBr. Round discs were obtained by pressing the ground material with the aid of a bench press and further readings were measured on NicoletTM 6700 model (ThermoScientific,USA) FTIR spectrometer. 2.2.2 Scanning electron microscopy combined with X-ray energy dispersive spectrometer (SEM-EDX) analysis The surface morphology and lead binding pattern in the lead-treated and the untreatedfungal biomasssampleswere recorded usingscanningelectron microscopycombinedwith X-ray energy dispersive spectrometer (SEM-EDX, JEOL, JSM 6490 LV, Japan). 2.3 Batch adsorption experiments Batch equilibrium experiments were performed with 20ml of a standard solution of concentration 5mg/l, 10mg/l and 15mg/l and a desired adsorbent dose of 0.1g to 1g. The parameters studied include initial metal ion concentration, contact time and adsorbent dose. The mixture was transferred to 50ml Erlenmeyer flasks and stirred for 2h to 6h, filtered using Whatman filter No. 42 and analyzed for residual metal concentration through inductively coupled plasma- mass spectrometry (ICP-MS). All the experiments were performed at room temperature conditions. The amount of lead adsorbed onto fungal biomass was calculated by using the following expression Eq. (1) (1) Where qe is the quantity of lead biosorbed onto fungal biomass as mg/g; V is the volume of lead solution inliters; Co and Cf are initial and final concentrations (mg/l) of lead in the solution and M is the weight of biomass taken for the batch experiment in g. 2.4 Adsorption isotherm Models The mathematical relationshipbetweentheconcentrationof adsorbate before and remained after the sorption experiment is explained by using adsorption isotherm models. Langmuir and Freundlich adsorption isotherm models are used in this study. These are commonly used to assess the sorption capacity of bio-adsorbent in the adsorption system. 2.4.1 Langmuir Isotherm It is generally based on monolayer adsorption process i.e., once all the sites available on adsorbent are filled, no more adsorption process can take place or say it reaches the saturation point. The general and the linearized form of this isotherm are represented in equations (2) and (3) respectivelyasfollows: (2) (3) Where qe is the amount of sorbed metal at equilibrium (mg/g), qmax is the monolayer sorption capacity (mg/g), b is the Langmuir constant, Ce is concentration of metal ions in solution at equilibrium.
  • 3. International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056 Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072 © 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7146 When a plot of (Ce/qe) versus Ce is plotted, a straight line with a slope of (1/qmax) and an intercept of (1/bqmax) must be obtained. 2.4.2 Freundlich isotherm Heterogenous surfaces involving adsorption processes are explained by using Freundlich isotherm. It is empirical and does not assume monolayer adsorption but is quiteuseful to explain adsorption data andfractionofactivessitesavailable for sorption study. The general and the linearized form of this isotherm are represented in equations (4) and (5) respectivelyasfollows: (4) (5) Where Ce is the equilibrium concentration (mg/g), qe is the adsorbed amount at equilibrium (mg/g),Kf istheadsorption capacity (L/mg) and 1/� is adsorption intensity, it also indicates the relative energy distribution and the heterogeneity of the adsorbate sites. 3. RESULTS AND DISCUSSION 3.1 Effect of initial metal ion concentration The effect of initial metal ion concentration was determined at operational conditions of 0.1g adsorbentdoseandcontact time of 2h at pH 5.5 for initial metal ionconcentrationfrom5 mg/l to 15 mg/l. Sorption capacity (qe) was found to be increased from 0.42 mg/g to 1.136 mg/g indicating an increase in adsorption rate with increase in initial metal ion concentration (Fig. 1) and reduced amount of metal ions in the aqueous solution. These findings could be explained as there is an acting huge force for rapid mass transfer, more the concentration of initial metal ions more will be adsorption rate.[18][19] Fig.1. Initial metal ion concentration Vs % Adsorption 3.2 Effect of contact time An investigation into the adsorption process in relation to contact time is an important parameter as it can help us to assess the efficiency of adsorption and to determine the saturation point with constant adsorbent dose and fixed initial metal concentration. Because at the beginning of the sorption process, more of the actives sites on the adsorbent is available and unoccupied and with the increase in time actives sites begins to get saturated by the metal ions present in the solution thereby decreasing the adsorption rate. [20] In the present study, the effect of contact time was determined at 10ppm/20ml with 0.1g adsorbent dose at pH 5.5 with contact time range from 2h to 6h and results showed decrease in concentration of metal ions in the solution and increased adsorption rate as up to 6 hours which can be explained due to unsaturation of actives sites and were still available for adsorption process (Fig. 2). Fig.2. Contact time Vs % Adsorption 3.3 Effect of adsorbent dose There is a direct relationship between adsorbent dose and percent adsorption. As we increase the dose of adsorbent and keep the initial metal ion concentration constant, more active sites are available for the metal ion to get adsorbed at the surface of adsorbent [21]. In the present study, the effect of adsorbent dose was determined at fixed 10ppm/20ml initial metal ion concentration and a contacttime of6hour at pH 5.5 and adsorbent dose ranges from 0.1g to 1g and results indicates increased adsorption rate as more actives sites are available with increased adsorbent dose at fixed concentration of metal ion (Fig. 3). Fig. 3 Adsorbent dose Vs % Adsorption
  • 4. International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056 Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072 © 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7147 3.4 SEM-EDX and FTIR analysis SEM-EDX analysis tells us about the surface morphologyand structural characterization of adsorbent along with their chemical composition and relative spectral energies.Results of lead untreated and treated adsorbentareshowninfig.4,5 and 6 respectively. An attempt was made to explore the surface of dry fungal adsorbent for any modification which could have been occurred due to adsorption of lead ions and fig. 4(b) indicates rough irregular morphology containing dead ruptured hyphae of fungal biomass loaded with lead ions in comparison to fig. 4(a) which is taken before adsorption of lead ions. EDX analysis as shown in fig. 5( before adsorption) and fig. 6( after adsorption) respectively revealed change in inorganic chemical constituents by weight percent and some reduction indicating ion-exchange could be considered as one of the dominant adsorption mechanism in lead ion biosorption onto fungal biomass. [22][23] (a) (b) Fig. 4 SEM micrographs of fungal adsorbent before sorption (a) and after sorption (b) of Pb2+ ions (15ppm) at 1000X magnification Fig. 5 EDX spectrum of Fungal adsorbent before adsorption Fig. 6 EDX spectrum of Fungal adsorbent after adsorption FTIR analysis is done to determine the types of functional groups present on the surface of the adsorbent. Results ofIR spectra before and after adsorption are presentedinfig.7(a) and 7(b) respectively. The largest peak of wave number 3384 cm-1 shown in Figure 7, corresponds to either free or H-bonded O-H groups that could be present in carboxylic acids on the surface of the fungal adsorbent. The peak at 2924.2 cm-1 wavenumbercorrespondstotheC-Hstretching that could exist in the alkanes. As for peaks ofwavenumbers in the range of 1650.2 to 1314.0 cm-1, are an indication of the olefinic and aromatic bond, and bands of wavenumber 1235.5 to 573.3 cm-1 are an indication of C-O bond. C-O bond may be due to carboxylic acids,hydroxygroupsorfiber carbonaceous that are present in the structure of fungal adsorbent that is composed of lignin and cellulose. Those groups probably act as proton donors that once get deprotonated, the hydroxyl group or the carboxyl group adsorb the heavy metal ions. As shown in Fig. 7(b), the peak representing the O-H bond was only shifted from 3384 cm-1 to 3392.3cm-1,whichisan insignificant change. Whereas the peak of the C=O bond was shifted from 573.3 to 607.0 cm-1, and the peak of the C-O bond was shifted from 1066.8 to 1107.8 cm-1. Those shifts can be attributed to the surface complexation created between Pb2+ and the carboxylic acids functional groups; hence, it is an indication that chemical complexation between Pb2+ and the carboxyl groups could be one of the mechanisms partially responsible for the biosorption of Pb2+ via fungal adsorbent. [24] (a)
  • 5. International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056 Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072 © 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7148 (b) Fig. 7 FTIR spectrum of fungal adsorbent before adsorption (a) and after adsorption (b) 3.5 Adsorption isotherms The values of Langmuir and Freundlich constants for lead ions uptake on dry fungal biomass are given in Table 1 and 2 respectively. The valuesofLangmuirconstantsqmaxand b are calculated by plotting against 1/qe and 1/Ce and Freundlich constants KF and n were obtained by the plot of log qe vs log Ce. The R2 value of Langmuir constant was 0.9984 wasfound to be higher than R2 value of Freundlich (0.9809), indicating that Langmuir data fitted well than Freundlich data in adsorption process (Figure 4 & 5).Itisconsideredto be well- fitted model if R2 value is close to unity. [25] Figure 4: Langmuir isotherm Table 1: Parameters of Langmuir isotherm qmax (mg/g) b(l/mg) R2 0.16 25.94 0.9984 Figure 5: Freundlich isotherm Table 2: Sorption Parameters of Freundlich isotherm Kf n R2 3.80 4.59 0.9809 4. CONCLUSIONS The present research was designed to investigate the efficiency of a dry fungal adsorbent for the removal of lead ions from their aqueous solutions. The effect of parameters such as initial metal ion concentration, contact time and the adsorbent dose was selected for adsorption study. The selected dead biomass of fungus showed a notable metal adsorption capacity over a limited range of parameters. The Langmuir adsorption isotherm was found to be best fitted with the present study. The dead fungal biomass may be employed in the future for heavy metal remediation from real wastewater and comparative study can be assessed between dead and living fungal biomass. Further, column studies could be done to compare metal uptake and sorption capacity with different metal ion concentrations, adsorbent dose and contact time. Column bed height could be varied to compare adsorption efficiency. 5. ACKNOWLEDGMENT Authors are thankful to the Head and Coordinator, Department of Microbiology, University of Lucknow, Lucknow for biomass availability and support. REFERENCES 1. Jacob, Jaya Mary, et al. Biological approaches to tackle heavy metal pollution: A survey of literature. Journal of environmental management, 2018; 217: 56-70.
  • 6. International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056 Volume: 06 Issue: 03 | Mar 2019 www.irjet.net p-ISSN: 2395-0072 © 2019, IRJET | Impact Factor value: 7.211 | ISO 9001:2008 Certified Journal | Page 7149 2. Zhao M, Xu Y, Zhang C, Rong H, Zeng G. New trends in removing heavymetalsfromwastewater.Applied microbiology and biotechnology. 2016; 100(15):6509-18. 3. Gupta A, Joia J, Sood A, Sood R, Sidhu C, Kaur G. Microbes as potential tool for remediation of heavy metals: a review. J Microb Biochem Technol. 2016; 8(4):364-72. 4. Rafiu Awofolu O, Okechukwu Okonkwo J, Badenhorst J, Jordaan E. A new approach to chemical modificationprotocolsofAspergillusniger and sorption of lead ion by fungal species. Electronic Journal of Biotechnology. 2006; 9(4):340-348. 5. Çabuk A, Ilhan S, Filik C, ÇALIŞKAN F. Pb2+ Biosorption by Pretreated Fungal Biomass. Turkish Journal of Biology. 2005; 29(1):23-28. 6. Dursun AY, Uslu G, Cuci Y, Aksu Z. Bioaccumulation of copper (II), lead (II) and chromium (VI) by growing Aspergillus niger. Process Biochemistry. 2003; 38(12):1647-51. 7. Akar T, Tunali S, Kiran I. Botrytis cinerea as a new fungal biosorbent for removal of Pb (II) from aqueoussolutions.Biochemical EngineeringJournal. 2005; 25(3):227-35. 8. Iqbal M, Edyvean RG. Biosorption of lead, copper and zinc ions on loofa sponge immobilized biomass of Phanerochaete chrysosporium. Minerals Engineering. 2004; 17(2):217-23. 9. Özer A, Özer D. Comparative study of the biosorption of Pb (II), Ni (II) and Cr (VI) ions onto S. cerevisiae: determination of biosorption heats. Journal of hazardous materials. 2003; 100(1- 3):219-29. 10. Yan G, Viraraghavan T. Heavy-metal removal from aqueous solution by fungus Mucor rouxii. Water research. 2003; 37(18):4486-96. 11. Iskandar NL, Zainudin NA, Tan SG. Tolerance and biosorption of copper (Cu) and lead (Pb) by filamentous fungi isolated from a freshwater ecosystem. Journal of Environmental Sciences. 2011; 23(5):824-30. 12. Skowroński T, Pirszel J, Skowrońska BP. Heavy metal removal by the waste biomass of Penicillium chrysogenum. Water Quality Research Journal. 2001; 36(4):793-803. 13. Niu H, Xu XS, Wang JH, Volesky B. Removal of lead from aqueous solutions by Penicillium biomass. Biotechnology and Bioengineering. 1993; 42(6):785-7. 14. Zeng X, Wei S, Sun L, Jacques DA, Tang J, Lian M, Ji Z, Wang J, Zhu J, Xu Z. Bioleaching of heavy metals from contaminated sediments by the Aspergillus niger strain SY1. Journal of Soils and Sediments. 2015; 15(4):1029-38. 15. Kumar R, Sharma AK, Singh P, Dhir B, Mehta D. Potential of some fungal and bacterial species in bioremediation of heavy metals. Journal of Nuclear Physics, Material Sciences, Radiation and Applications. 2014; 1:213-223. 16. Mustapha MU, Halimoon N. Microorganisms and biosorption of heavy metals in the environment: a review paper. J. Microb. Biochem. Technol. 2015; 7:253-6. 17. Ayangbenro A, Babalola O. A new strategyforheavy metal polluted environments: a review of microbial biosorbents. International journal ofenvironmental research and public health. 2017; 14(1):94. 18. Pandhare G, Trivedi N, Pathrabe R, Dawande SD. Adsorption of cadmium (II) and lead (II) from a stock solution using neem leaves powder as a low- cost adsorbent. International Journal of Innovative Research in Science, Engineering and Technology. 2013;2(10). 19. Oves M, Khan MS, Zaidi A. Biosorption of heavy metals by Bacillus thuringiensis strain OSM29 originating from industrial effluent contaminated north Indian soil. Saudi journal of biological sciences. 2013; 20(2):121-9. 20. Fadel M, Hassanein NM, Elshafei MM, Mostafa AH, Ahmed MA, Khater HM. Biosorption of manganese from groundwater by biomass of Saccharomyces cerevisiae. Hbrc Journal. 2017; 13(1):106-13. 21. Rezaei H. Biosorption of chromium by using Spirulina sp. Arabian Journal of Chemistry. 2016; 9(6):846-53. 22. Gola D, Dey P, Bhattacharya A, Mishra A, Malik A, Namburath M, Ahammad SZ. Multiple heavy metal removal using an entomopathogenic fungi Beauveria bassiana. Bioresource technology. 2016; 218:388-96. 23. Pan X, Wu W, Lü J, Chen Z, Li L, Rao W, Guan X. Biosorption and extraction of europium by Bacillus thuringiensis strain. Inorganic Chemistry Communications. 2017; 75:21-4. 24. Selatnia A, Bakhti MZ, Madani A, Kertous L, Mansouri Y. Biosorption of Cd2+ from aqueous solution by a NaOH-treated bacterial dead Streptomyces rimosus biomass. Hydrometallurgy. 2004; 75(1-4):11-24. 25. Ayawei N, Ebelegi AN, Wankasi D. Modelling and interpretation of adsorption isotherms. Journal of Chemistry. 2017; 2017.