Bioresource Technology 184 (2015) 395–404

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Defatted algal biomass as a non-conventional low-cost adsorbent:

Surface characterization and methylene blue adsorption characteristics
T. Sarat Chandra a,c,d, S.N. Mudliar d, S. Vidyashankar d, S. Mukherji c, R. Sarada d, K. Krishnamurthi b,
V.S. Chauhan d,⇑
a
Environmental Biotechnology Division, CSIR-National Environmental Engineering Research Institute, Nagpur, Maharashtra, India
b
Environmental Health Division, CSIR-National Environmental Engineering Research Institute, Nagpur, Maharashtra, India
c
Centre for Environmental Science and Engineering, Indian Institute of Technology Bombay, Mumbai, Maharashtra, India
d
Plant Cell Biotechnology Department, CSIR-Central Food Technological Research Institute, Mysore, Karnataka, India

h i g h l i g h t s

 Utilization of defatted algal biomass (DAB) as an adsorbent was studied.

1
 Maximum adsorption capacity of DAB was found to be 7.73 mg g .
2 1
 Specific surface area of DAB was estimated to be 18.94 m g .

a r t i c l e i n f o a b s t r a c t

Article history: The present study investigates the use of defatted algal biomass (DAB) as a non-conventional low cost
Received 5 August 2014 adsorbent. The maximum adsorption capacity of biomass (raw, defatted and sulfuric acid pretreated
Received in revised form 1 October 2014 DAB) was determined by liquid phase adsorption studies in batch mode for the removal of methylene
Accepted 4 October 2014
blue present at various concentrations (1, 2, 3, 4, and 5 mg L1) from aqueous solutions. The data was well
Available online 13 October 2014
fitted with Langmuir and Freundlich isotherms. The maximum adsorption capacity for raw, defatted and
sulfuric acid pretreated DAB was found to be 6.0, 7.73 and 7.80 mg g1, respectively. The specific surface
Keywords:
area of raw, defatted and sulfuric acid pretreated DAB was estimated to be 14.70, 18.94, and 19.10 m2 g1,
Defatted algal biomass
Langmuir and Freundlich adsorption
respectively. To evaluate the kinetic mechanism that controls the adsorption process, pseudo-first order,
isotherm pseudo-second order, intraparticle diffusion and particle diffusion has been tested. The data fitted quite
Methylene blue well with pseudo-second order kinetic model.
Specific surface area Ó 2014 Elsevier Ltd. All rights reserved.
Kinetics

1. Introduction production of methane through anaerobic digestion (Yen and

Oleaginous microalgal biomass is recognized as a potential feed
stock for production of biofuels. Despite many advantages, the
commercialization of algal biofuels faces enormous challenges
due to its high processing costs and energy requirements. About
70% of the whole algal biomass remains as the defatted algal bio-
mass (DAB) after the extraction of lipids for biofuel application
(Park et al., 2012). A proper economic utilization of the defatted
algal biomass (DAB) may improve the overall process economics
of algal biofuel production. It can be used as an alternative food
supplement for livestock (Howe et al., 2010) and also for
⇑ Corresponding author.
E-mail address: vikas@cftri.res.in (V.S. Chauhan).
Brune, 2007; Ehimen et al., 2009; Park et al., 2009; Suresh et al.,
2013; Sarat Chandra et al., 2014). Another value-added application
of DAB could be as adsorbent for removal of heavy metals and dyes
from industrial wastewaters. Biogas production from microalgal
biomass requires pretreatment, while, use of DAB as adsorbent
does not require any pretreatment and thus does not pose any eco-
nomic burden (Rashid et al., 2013). Coal-based activated carbon is
the most commonly used adsorbent for the removal of dyes
because of its high surface area and adsorption capacity. There is
growing interest in search of alternative for carbon-based
adsorbents as these are derived from non-renewable sources
(Tseng et al., 2006) and are relatively expensive. Usage of organic
carbon as adsorbent is advantageous as they are usually derived
from renewable resources like biomass which are abundantly

http://dx.doi.org/10.1016/j.biortech.2014.10.018
0960-8524/Ó 2014 Elsevier Ltd. All rights reserved.
396 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404

available after industrial processes as residue, byproducts and treated biomass was further soaked in 2% NaHCO3 solution to
waste and hence are comparatively inexpensive (Forgacs et al., remove any remaining acid and the pH was adjusted to 7. The
2004). Microalgal cell wall provides binding sites such as hydroxyl biomass was dried in an oven at 150 °C for 46 h under nitrogen
and carboxyl groups for several basic dyes that exist as cations in flow (Esmaeili and Ghasemi, 2009).
aqueous solution. As the mode of uptake is extracellular, the func-
tional groups on the cell wall play an important role in adsorption
system. Utilization of macroalgal and filamentous microalgal bio- 2.4. Adsorption studies
mass as an adsorbent is widely reported (Vilar et al., 2007; Gupta
and Rastogi, 2008a,b,c, 2009; Esmaeili and Ghasemi, 2009; The adsorption studies were carried out in a 2 L Erlenmeyer
Aravindhan et al., 2009; Bekci et al., 2009). Studies on utilization flask. The methylene blue solution was prepared with deionized
of unicellular microalgae for adsorption have been limited water. The adsorption of methylene blue by DAB was investigated
(Cetinkaya et al., 1999; Schmitt et al., 2001; Tsai and Chen, 2010; for the initial dye concentration range 1-5 mg/L at adsorbent dose
Tsai et al., 2011; Mishra and Mukherji, 2012). Cetinkaya et al. of 1 g L1. The concentrations of dye and the dose of adsorbent
(1999) reported the use of Scenedesmus obliquus and Synechocystis were chosen based on earlier studies with similar system (Tsai
sp. for adsorption of copper, nickel and chromium. Schmitt et al. et al., 2011). The initial pH of the dye solution was adjusted to
(2001) investigated the use of four live microalgae namely Scene- 7.0 by using dilute HCl solution. The flasks were stirred at
desmus subspicatus, Cyclotella cryptica, Phaeodactylum tricornutum, 400 rpm (rotations per minute) using a magnetic stirrer. The tem-
Porphyridium purpureum for adsorption of six metal ions namely perature of dye solution during adsorption studies was maintained
aluminium, zinc, mercury, lead, copper, and cadmium. Tsai and at 25 °C. The samples were withdrawn at regular intervals up to 1 h
Chen (2010) and Tsai et al. (2011) examined the use of live and and were filtered using mixed cellulose ester (MCE) membrane of
dead Chlorella biomass, as an effective low cost adsorbent for pore size 0.45 lm. The dye concentration in the filtered solution
removal of malachite green and methylene blue, respectively, from was measured using a UV/Visible spectrophotometer (Model:
aqueous solutions. Mishra and Mukherji (2012) reported the use of UV/1800A, Shimadzu corporation, Kyoto, Japan) at a wavelength
dead Scenedesmus abundans and Spirulina biomass as a low cost of 663.8 nm (maximum absorbance was observed at this wave-
adsorbent for adsorption of diesel and lubricating oil. length). The amount of adsorption at time t, qt (mg g1) was deter-
However, the use of defatted algal biomass as adsorbent has lar- mined by,
gely remained unexplored. The Scenedesmus dimorphus strain, used
in the present study, has been reported to have high CO2 tolerance ððC o  C t Þ  VÞ
qt ¼ ð1Þ
(15% CO2 v/v), faster growth and moderate lipid accumulation W
(>20% w/w) (Vidyashankar et al., 2013). The present study evalu-
ates the defatted biomass of S. dimorphus strain for its potential where, Co and Ct (mg L1) are the concentrations of methylene blue
as a non-conventional adsorbent and the surface characteristics at initial and at any time t, respectively. V is the volume of the solu-
of DAB were studied using methylene blue as adsorbate. The effect tion (L) and W is the mass of dry adsorbent used (g).
of sulfuric acid pretreatment on adsorption capacity of DAB was
also studied. Further, adsorption isotherms and kinetic models Table 1
FTIR analysis of raw, DAB, and sulfuric acid pretreated DAB.
were tested to understand the mechanisms that control the
adsorption process. Sample Stretching Assignments
frequencies (cm1)
Raw algal biomass 3513, 3417, 3379, –OH stretch of alcohols/–NH
2. Methods
3330 stretch
3303, 3179 –OH stretch of alcohols
2.1. Microalgal biomass 1585 –NH bend
1412 –OH bending of alcohol/
Biomass of an oleaginous microalga, S. dimorphus, was –COOH
1107 –CN stretch of amine
harvested from cultures growing in outdoor open raceway ponds 855, 808, 780, 752, –C@C bending
in phototrophic bold basal medium and sun dried. The lipids were 719, 694
extracted from the dry biomass (moisture content 6% (w/w)) 625, 581, 539, 515 –C–Cl
with hexane using soxhlet apparatus. The DAB was sun dried and DAB 3639, 3524, 3462, –OH stretch of alcohols/–NH
used for further studies. 3412 stretch
3278 –OH stretch of alcohol
1584 –NH bend
2.2. Biomass characterization 1408 –OH bending of alcohol/–COOH
1090, 1052 –COOH
The surface morphology of biomass samples were examined 854, 826, 746 –C@C bending
720 –C–H bend
using a LEO Scanning Electron Microscope 435 VP (Leo Electron
657, 589, 560, 518 –C–Cl
Microscopy Ltd., Cambridge, UK). FTIR analysis was performed
Sulfuric acid 3744 –OH stretch of alcohol
using Fourier Transform Infra-red Spectrometer (FTIR, Model: IFS
pretreated DAB
25, BRUKER Germany). 3425 –NH stretch of amine
2228 –CN stretch of nitrile/alkyne
disubstituted
2.3. Sulfuric acid pretreatment
2135 –CN stretch of nitrile/alkyne
monosubstituted
The defatted algal biomass (100 g) was added to 100 mL of 97% 1660 –C@C stretch trisubstituted
H2SO4 in a fume cupboard. The resultant mixture was kept for 24 h 1624 –C@C stretch conjugated alkene
at room temperature, and was subsequently refluxed in the fume 1464 –CH bend alkane
798 –C@C bend trisubstituted
hood for 4 h. The mixture was allowed to cool to room tempera- 676 –C@C bend disubstituted
ture. The acid layer was removed, and the biomass thus treated 614, 595 –C–Cl
was washed several times with deionized water. The resultant
 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404 397

0.35

0.3 (a)
0.25

0.2

Ct/Co
0.15

0.1

0.05

0
0 10 20 30 40 50 60 70
t (min)

0.18

0.16 (b)
0.14

0.12

0.1
Ct/Co

0.08

0.06

0.04

0.02

0
0 10 20 30 40 50 60 70
t (min)

0.7
(c)
0.6

0.5

0.4
Ct/Co

0.3

0.2

0.1

0
0 10 20 30 40 50 60 70
t (min)

Fig. 1. The removal of methylene blue from aqueous solution onto the raw (a), DAB (b) and sulfuric acid pre-treated defatted (c) algal biomass (d 1 mg L1; } 2 mg L1; N
3 mg L1; j 4 mg L1;  5 mg L1).

The amount of dye adsorbed at equilibrium, qe (mg g1) was 3. Results and discussion
determined by,
3.1. Characterization of raw, defatted and sulfuric acid pre-treated
defatted algal adsorbent
ððC o  C e Þ  VÞ
qe ¼ ð2Þ
W Particle density, organic matter and ash content of raw algal
biomass (RAB), defatted algal biomass (DAB), and sulfuric acid pre-
where, Ce (mg L1) is the concentration of methylene blue at treated DAB was carried out. The organic matter (Volatile solids) in
equilibrium. raw algal biomass, defatted, and sulfuric acid pretreated DAB was
All the experimental values indicated in the graphs were 82.28%, 70.56% and 58.86%, respectively. The ash content was
based on two sets of observations and with standard 6.98%, 21.05%, and 38.34%, respectively, for raw, defatted, and sul-
deviation < 10%. furic acid pretreated DAB. The particle density of the raw, defatted
398 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404

5
4.5 (a)
4
3.5

qe(mg g-1)
3
2.5
2
1.5
1
0.5
0
0 0.1 0.2 0.3 0.4 0.5 0.6
Ce (mg L-1)
Expermental Data Langmuir Freundlich

6
(b)
5

4
q e(mg g-1)

0
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45
Ce (mg L-1)
Experimental Data Langmuir Freundlich

4.5
4 (c)
3.5
3
q t (mg g-1)

2.5
2
1.5
1
0.5
0
0 0.2 0.4 0.6 0.8 1 1.2
Ce (mg L-1)

Experimental data Langmuir Freundlich

Fig. 2. Adsorption isotherm of methylene blue in aqueous solution onto raw, defatted, and sulfuric acid pretreated defatted algal biomass.

and sulfuric acid pretreated DAB adsorbent was 0.82 g mL1,
0.85 g mL1, 0.85 g mL1, respectively. The surface morphology of
raw, defatted, and sulfuric acid pre-treated defatted algal biomass
was characterized by Scanning Electron Microscopy (SEM) imaging
(magnification of 10,000). The SEM images showed that as the
algal biomass undergoes pre-treatment affecting the cell wall,
i.e., process of solvent extraction for defatting and further
treatment with sulfuric acid, the surface texture becomes more
corrugated with progressive enhancement in surface roughness.
The raw algal cells showed a spherical and comparatively compact
and smooth surface compared to the defatted and sulfuric acid pre-
treated defatted algal biomass. The cells of defatted algal biomass
appeared deformed with some amount of disintegration. The max-
imum cell deformation, corrugation and roughness with visibly
enhanced surface porosity were shown by the sulfuric acid pre-
treated defatted algal biomass.
The functional groups present on the surface of the raw, defat-
ted and sulfuric acid pre-treated DAB were identified by analysis of
the FTIR spectrum based on the reported spectral analysis of algae
(Mishra and Mukherji, 2012; Gupta and Rastogi, 2009) and are
given in Table 1. The data indicates that the surface of raw, and
DAB is populated with negatively charged functional groups
(mainly –OH and –COOH). The complex nature of adsorbent is
displayed by the presence of a number of absorption peaks in
raw, defatted and sulfuric acid pretreated DAB. Pretreatment with
sulfuric acid led to the disappearance of some of the peaks present
in DAB, and new peaks were observed in FTIR spectra indicating an
increase in C@C and C„N functional groups (Table. 1). There are no
 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404 399

reported case studies with microalgae, however studies on equiv- Table 2

alent materials like plant biomass (e.g., Euphorbia rigida) reported Comparison of adsorption capacities of various adsorbents for methylene blue.

that sulfuric acid pretreatment lead to aromatization and carbon- Adsorbent Maximum monolayer References
ization which was indicated via C@C stretching frequencies in FTIR adsorption capacity (mg g1)
spectra (Gercel et al., 2007). Oak sawdust 29.94 Ferrero (2007)
Pitch-pine sawdust 27.78 Ferrero (2007)
Glass wool 2.24 Chakrabarti and
3.2. Adsorption characteristics of methylene blue onto raw, defatted Dutta (2005)
and sulfuric acid pre-treated defatted algal biomass Gelidium algae 64 Vilar et al.
(2005)
The removal of methylene blue from aqueous solution onto the Spirogyra sp. 140.84 Gupta and
Rastogi (2008a)
raw, DAB and sulfuric acid pre-treated DAB over time are shown in Activated carbon from 8.82 Aygun et al.
Fig. 1. It is obvious from the figure that rapid decrease in solution hazelnut shell (2003)
phase concentration observed initially was followed by a slow Activated carbon from 114.45 Gercel et al.
decrease beyond 20 min in raw and defatted algal adsorbent. Lev- Euphorbia rigida (2007)
Activated carbon from 70.92 Hameed et al.
elling off was observed beyond 30 min in sulfuric acid pretreated
coconut bunch waste (2008)
defatted algal adsorbent. This may be due to the saturation of neg- Scenedesmus sp. 6.0 Present study
ative charges present on the surface of the adsorbent. Also, the ini- Defatted Scenedesmus sp. 7.73 Present study
tial slower rate of adsorption noticed with sulfuric acid pretreated biomass
DAB may be attributed to changes in the functional groups present Sulfuric acid pretreated 7.80 Present study
Scenedesmus DAB
on the surface as a result of acid pretreatment. The initial pH of
methylene blue solution of different concentration (1–5 mg L1)
were in the alkaline range (pH 7.3–7.8) and were adjusted to
neutral pH for the adsorption studies. The final pH values at the
Although the adsorption capacity of DAB is lower compared to
equilibrium were observed to be in the acidic range (pH 6–6.5)
commonly used adsorbents such as activated carbon, the abun-
in various experiments with different adsorbents. Two common
dance of availability due to a growing microalgal industry and
isotherm equations namely Langmuir and Freundlich equations
low processing costs makes DAB a promising source of adsorbent.
(Vilar et al., 2007) were applied to model the isotherms in the
present study.
  3.3. Determination of specific surface area of raw, defatted, and
1 1 1 1 pretreated DAB
Langmuir : ¼  þ ð3Þ
qe ½ðK L  qm Þ Ce qm
 
1 The prerequisite of degassing the biomass for N2 adsorption in
Freundlich : lnðqe Þ ¼ lnðK F Þ þ lnðC e Þ ð4Þ
n Brunauer–Emmett–Teller (BET) method has the potential to
disturb the surface properties and may lead to unreliable results.
where, Ce, qe are the concentration of the dye in solution (mg L1) Therefore, adsorption of dissolved molecules, like methylene blue,
and the loading of the dye on the adsorbent (mg g1) at equilibrium, from aqueous solution and fitting with the Langmuir isotherm pro-
respectively, KL is a direct measure of intensity of the adsorption vides an alternative method for estimating specific surface area of
process (mL mg1), qm is the Langmuir maximum monolayer porous solids (Vilar et al., 2007). Assuming monolayer adsorption,
adsorption capacity (mg g1), KF (mg1n Lng1) and n are the the surface area of adsorbent per unit mass (As, m2 g1) can be
Freundlich isotherm constants. 1/n is a measure for the surface related to the monolayer adsorption capacity (qm) by the following
heterogeneity and 1/n values lesser than 1 indicate a higher adsorp- equation.
tion intensity. From a linear fit to sorption isotherm data plotted on
1/qe vs. 1/Ce, qm and KL can be determined based on the slope and 103  qm  NA  Am
As ¼ ð5Þ
the intercept. Similarly, KF and 1/n can be determined from the M
intercept and slope of a linear fit to sorption data plotted on ln
(qe) vs. ln (Ce) axes. where NA is the Avogadro’s number (NA = 6.023 * 1023 molecules
The Langmuir and Freundlich isotherms at 25 °C for raw, DAB mol1), Am being effective molecular area for methylene blue
and sulfuric acid pre-treated DAB are shown in Fig. 2. Both models (Am = 1.3 * 1018 m2) (Vilar et al., 2007) and M is the molecular
provided good fit to data (Table 3). The adsorption isotherms weight for methylene blue (M = 319.89 g mol1). The qm observed
revealed that the Langmuir adsorption capacity of raw algal bio- for raw algal biomass, DAB and sulfuric acid pretreated DAB was
mass and DAB was 6.0 mg g1 and 7.73 mg g1, respectively, for 6.0, 7.73 and 7.80 mg g1, respectively. Therefore, the value of As
the basic dye in an aqueous medium. Tsai et al. (2011) reported for raw algal biomass, DAB and sulfuric acid pretreated DAB was
the adsorption capacity of methylene blue to be around 5.0 mg g1
for dead biomass of Chlorella sp. The sulfuric acid pretreated DAB Table 3
showed enhanced adsorption capacity with Langmuir monolayer Langmuir and Freundlich isotherm model constants and correlation coefficients for
adsorption capacity of 7.80 mg g1 for the basic dye from an aque- adsorption of methylene blue onto raw, DAB and sulfuric acid pretreated DAB.
ous medium. This marginal enhancement in dye uptake may be Isotherm Raw algal biomass DAB Sulfuric acid
due to the marginal enhancement in surface corrugation and pretreated DAB
porosity as exhibited by SEM imaging. Also, the 1/n values of Langmuir
RAB, DAB and sulfuric acid pre-treated DAB in the present study KL (L mg1) 5.08 4.98 1.10
were less than 1, clearly indicating the algal biomass as a favorable qm (mg g1) 6.0 7.73 7.80
adsorbent. These results clearly indicate that DAB could be used as R2 0.94 0.93 0.99

an adsorbent for removal of basic dyes like methylene blue from Freundlich
aqueous solutions. The adsorption capacity of DAB is therefore KF (mg1-n Lng1) 6.20 7.86 4.34
1/n 0.52 0.53 0.70
comparable to or superior to various agricultural waste, microalgal
R2 0.94 0.91 0.97
waste and aquaculture waste based potential adsorbents (Table 2).
400 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404

70
(a) 60 (A)
50

t/qt (min g mg-1)

1/qt (g mg-1)
40

30

20

10

0
0 0.05 0.1 0.15 0 20 40 60 80
1/t (min-1) t (min)

1.4 70

1.2 (b) 60 (B)

t/qt (min g mg-1)

1 50
1/qt (g mg-1)

0.8 40

0.6 30

0.4 20

0.2 10

0 0
0 0.05 0.1 0.15 0 20 40 60 80
t (min)
1/t (min-1)

(c) (C)
t/qt (min g mg-1)
1/qt (g mg-1)

1/t (min-1) t (min)

Fig. 3. The pseudo first order and second order kinetic plots for adsorption of methylene blue at various concentrations onto raw (a and A), defatted (b and B) and sulfuric acid
pretreated defatted (c and C) algal biomass (d 1 mg L1; } 2 mg L1; N 3 mg L1; j 4 mg L1;  5 mg L1).

determined as 14.70 m2 g1, 18.94 m2 g1 and 19.10 m2 g1,
respectively.
3.4. Adsorption kinetics
Pseudo-first order, pseudo-second order, particle diffusion, and
intraparticle diffusion models were tested to assess the kinetic
mechanism that controls the adsorption process. The kinetic mod-
els for methylene blue adsorption onto raw, defatted, and sulfuric
acid pre-treated biomass are shown in Figs. 3–5. The kinetic data
for the adsorption of methylene blue onto raw defatted and sulfu-
ric acid pretreated defatted algal biomass were calculated from the
related plots and are given in Tables 4–6.
The pseudo-first order kinetic model equation (Kannan and
Sundaram, 2001) is:
   
1 1 k1 1
¼ þ  ð6Þ
qt q1 q1 t
where, q1 is the amount of dye adsorbed at equilibrium expressed in
mg g1 based on the pseudo first order model, and k1 is the pseudo
first order rate constant (min1). Based on the pseudo first order
model, these parameters were determined from a linear fit of sorp-
tion rate data, as shown in Eq. (6), i.e., 1/qt vs. 1/t plot using the
slope and intercept values.
The pseudo-second order kinetic model equation (Ho and
Mckay, 1998) can be given as:
 
t 1 1
¼ 2
þ t ð7Þ
qt k2 q2 q2
where, q2 is the maximum adsorption capacity (mg g1) for the
pseudo-second order model and k2 is the pseudo second order rate
constant (g mg1 min1). Values of q2 and K2 were calculated from
the slope and intercept of the linear fits to sorption rate data plotted
on t/qt vs. t axes.
The coefficient of determination (R2) for the pseudo first order
kinetic model for raw, defatted, and sulfuric acid pretreated defat-
ted algal biomass as adsorbent were calculated and compared to
the second order kinetic model. It was observed that the correla-
tion coefficients for raw, defatted, and sulfuric acid pretreated
 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404 401

5
4.5
(a)
4
3.5

q t(mg g-1)
3
2.5
2
1.5
1
0.5
0
0 1 2 3 4 5 6 7 8 9
t^0.5 (min^0.5)

5
4.5 (b)
4
3.5
3
2.5
2
qt

1.5
1
0.5
0
0 1 2 3 4 5 6 7 8 9
t^0.5 (min^0.5)

5
4.5
4 (c)
3.5
q t(mg g-1)

3
2.5
2
1.5
1
0.5
0
0 1 2 3 4 5 6 7 8 9
t^0.5 (min^0.5)

Fig. 4. The intra-particle diffusion plots for adsorption of methylene blue at various concentrations onto raw (a), defatted (b) and sulfuric acid pretreated (c) defatted algal
biomass (d 1 mg L1; } 2 mg L1; N 3 mg L1; j 4 mg L1;  5 mg L1).

defatted algal biomass were comparatively lower for the pseudo
first order kinetic model (Tables 4–6). This implies that kinetics
of methylene blue adsorption by raw, defatted and sulfuric acid
pre-treated defatted algal biomass is better described by the
pseudo-second order kinetic model. The rate constants, k1 and k2,
varied with initial concentration of the dye. Similar variations were
observed by Hameed et al. (2008) for adsorption of methylene blue
by coconut coir waste.
The particle diffusion model (Weber and Morris, 1987) can be
represented as:
 
ct
ln ¼ kp  t
ce
where kp (min1) is the particle diffusion coefficient. The value of kp
can be obtained from the slope of the plot, ln (Ct/Ce) vs. t.
The intraparticle diffusion model (Crank, 1975) can be given by
the following equation:
qt ¼ kpi t0:5 þ c ð9Þ
where c is the intercept and kpi is the intraparticle diffusion rate
constant (mg g1 min0.5). According to this model, the plot of the
uptake, qt, vs. the square root of time, t1/2 should be linear if
intraparticle diffusion is involved in the process of adsorption. If
the line passes through the origin, then intraparticle diffusion is
the primary rate-controlling step. If the line does not pass through
the origin, then it suggests that intraparticle diffusion is not the sole
ð8Þ rate limiting step. In the present study for raw, defatted, and
sulfuric acid pre-treated algal biomass as adsorbent, the intraparti-
cle diffusion plots for the adsorbents were all linear, however, the
lines did not pass through the origin; thereby indicating that
402 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404

0
0 10 20 30 40 50 60 70
-0.5
(a)
-1
-1.5
Ln(Ct)
-2
-2.5
-3
-3.5
-4
t (min)

0
0 10 20 30 40 50 60 70
-0.5
(b)
-1

-1.5
Ln(Ct)

-2

-2.5

-3

-3.5

-4
t (min)

1.5

1
(c)
0.5

0
Ln(Ct)

0 10 20 30 40 50 60 70
-0.5

-1

-1.5

-2

-2.5
t (min)

Fig. 5. The particle diffusion plots for adsorption of methylene blue at various concentrations onto raw (a), defatted (b) and sulfuric acid pretreated (c) DAB (d 1 mg L1; }
2 mg L1; N 3 mg L1; j 4 mg L1;  5 mg L1).

intraparticle diffusion was not the only rate controlling step
(Hameed et al., 2008).
The R2 values for the particle diffusion model for raw, defatted,
and sulfuric acid pretreated defatted algal biomass as adsorbent
were calculated and compared to those determined for the
intraparticle diffusion model. The R2 values for particle diffusion
and intraparticle diffusion were almost equal for both raw and
defatted algal biomass, thereby implying that both the resistances
are contributing to adsorption of the dye. While, for sulfuric acid
pretreated defatted algal biomass the correlation coefficients for
particle diffusion are more than intraparticle diffusion. This may
be due to extensive corrugation and generation of porosity caused
during sulfuric acid pre-treatment as observed by SEM imaging.
The process of solvent extraction for defatting enhanced the
corrugation, roughness and porosity but not to the extent caused
during sulfuric acid pretreatment.
The mechanism of adsorption in the present study appears to
be primarily a function of ionic/electrostatic interaction between
the negatively charged surface of adsorbents and positively
charged dye molecule. Similar observations have been made
by Tsai et al. (2011) with dead Chlorella biomass. A marginal
enhancement in adsorption capacity on pretreatment with sulfu-
ric acid suggests that the enhancement in surface corrugation
and porosity may have only minor effect on overall adsorption
capacity of DAB. However, further studies on optimisation of
sulfuric acid pretreatment of microalgal biomass are required
to have a detailed understanding of its effect on adsorption
capacity.
 T. Sarat Chandra et al. / Bioresource Technology 184 (2015) 395–404 403

Table 4 of basic dyes from aqueous solution. The presence of negatively

Kinetic parameters for adsorption of methylene blue onto raw algal biomass. charged functional groups on adsorbent’s surface, as indicated by
Concentration (mg L1) FTIR analysis, contribute to the adsorption of dye at neutral pH
5 4 3 2 1 due to electrostatic interactions. The adsorption capacity and the
specific surface area of DAB was estimated to be 7.73 mg g1 and
Pseudo First-order
k1 (min1) 0.45 0.55 0.47 1.25 4.80
18.94 m2 g1 respectively.
q1 (mg g1) 4.52 3.58 2.88 1.93 0.98
R2 0.95 0.95 0.96 0.94 0.97
Acknowledgements
Pseudo second order
1/k2 (g1 mg min) 4.43 3.37 2.88 1.92 0.99
q2 (mg g1) 4.89 3.9 2.94 2.00 1.00 Financial support (BT/PR1311/PBD/26/267/2011) from Depart-
R2 0.99 0.99 0.99 0.99 0.99 ment of Biotechnology (DBT), Government of India is gratefully
Intraparticle diffusion acknowledged. TSC is thankful to Council of Scientific and Indus-
kpi (mg g1 min0.5) 0.03 0.03 0.02 0.04 0.06 trial Research (CSIR) for providing research fellowship.
R2 0.85 0.86 0.82 0.91 0.97
Particle diffusion
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