Science of the Total Environment 755 (2021) 142654

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Bio-sorption of toxic metals from industrial wastewater by algae strains

Spirulina platensis and Chlorella vulgaris: Application of isotherm, kinetic
models and process optimization
Fares Almomani ⁎, Rahul R. Bhosale
Department of Chemical Engineering, College of Engineering, Qatar University, P. O. Box 2713, Doha, Qatar

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Acidic treatment (Ac-T) enhanced toxic

metal (TMs) removal ability by algae
strains.
• Ac-T reformed surface functional groups
and increased TMs the removal effi-
ciency.
• Response surface methodology pre-
dicted optimum conditions for maxi-
mum %TMrem.
• The biosorption mechanism were a
combined electrostatic attraction and
ion-exchange.
• Up to 95% TM removal efficiency was
achieved by treated Spirulina platensis.

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

Article history: The present study evaluates the effect of an acidic treatment on the improvement of the percentage removal of
Received 27 May 2020 toxic metal (%TMrem) from wastewater by algae strains (Spirulina platensis (SP) and Chlorella vulgar (CV))
Received in revised form 1 September 2020 under different adsorbent dosages (0.2–2.5 g), a pH of (4–8) and contact time (5–100 min). The acidic treatment
Accepted 25 September 2020
(Ac-T) altered the functional groups on the surface of algae promoting more electronegative groups and im-
Available online 1 October 2020
proved the %TMrem of Al, Ni and Cu. Treated SP removed up to 95.0 ± 0.3% (Std. Dev = 0.24), 87.0 ± 0.2%
Guest Editor: Abdul Ghani Olabi (Std. Dev = 0.34)%, and 63.0 ± 0.3% (Std. Dev = 0.14) of Al, Ni, and Cu at the optimum pH of 5.5, 6.0, and, 7.0
and an adsorbent dosage of = 2.5 ± 0.1 g/L (Std. Dev = 0.14) g/L, respectively. Lower %TMrem of 87.0% ± 0.2
Keywords: (Std. Dev = 0.09), 79.1 ± 0.4% (Std. Dev = 0.08), and 80.0 ± 0.2% (Std. Dev = 0.04) were achieved with treated
Isotherms CV, respectively. The optimum operational conditions for maximum %TMrem were determined at (Calgae = 4.8 ±
Thermodynamic model 0.2 gMNPs.L−1, Ct = 88 ± 1, and pH = 6) using the response surface methodology (RSM). The adsorption of TMs
Treated bio-sorbent on algae is endothermic, spontaneous, and follows Langmuir and second-order kinetics. Zeta potential measure-
Spirulina platensis ments indicated that the adsorption mechanism between the toxic metal (TM) and algal strains is controlled by
Chlorella vulgar
electrostatic interaction. As such, bio-sorption is a sustainable and efficient technology for the removal of TM
from wastewater.
© 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://
creativecommons.org/licenses/by/4.0/).

1. Introduction

The industrial revolution, unprecedented industrial production, and

⁎ Corresponding author. the growth of urbanization have greatly increased the water demand,
E-mail address: falmomani@qu.edu.qa (F. Almomani). and has subsequently generated large volumes of contaminated

https://doi.org/10.1016/j.scitotenv.2020.142654
0048-9697/© 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
F. Almomani and R.R. Bhosale
wastewater (Weis, 2010; Rezaei and Sayadi, 2015; Karakurt et al., 2019;
Kim et al., 2019). Industrial effluents contaminated with metallic ions
from different industries (e.g. metal, manufacturing of electronics, min-
ing, and electroplating) are considered a serious threat to the environ-
ment (Yuan et al., 2018; Almomani et al., 2019a; Rahman and Singh,
2019; Siddiqui et al., 2019; Zhong et al., 2019; Jawed et al., 2020). The
presence of toxic metal ions such as nickel, aluminum, and copper in
water or wastewater, even at low concentrations, can cause serious
health issues for both human and biotic organisms (Sayadi et al.,
2015; Shen et al., 2020; Rasheed et al., 2018; Yan et al., 2020). Nervous
system damage, kidney failure, and cancer cases were connected to
human exposure to these TMs. Therefore, their treatment and removals
from water resources are crucial (Dahiya et al., 2008; Shin et al., 2011;
Visa, 2016; Ghaemi et al., 2017).
Conventional treatment processes such as chemical precipitation,
ion exchange, and membrane separation were successfully used for
TM removals (Efome et al., 2019; Ibrahim et al., 2019; Garba et al.,
2020). However, these processes are inefficient and economically un-
feasible (Dąbrowski et al., 2004; Kavand et al., 2017; Khan and Rao,
2017; Atari et al., 2019; Crini et al., 2019; Giwa et al., 2019). More re-
cently, bio-sorption has emerged as an alternative and promising solu-
tion for the removal of TMs (Khan and Khan Rao, 2017; Jokar et al.,
2019; Saavedra et al., 2019; Salama et al., 2019; Apiratikul, 2020). Bio-
sorbents are made of plant-based materials, agricultural waste, algae,
and/or bacteria that can uptake different contaminants from the waste-
water. Wheat bran, wheat straw, and green algae were reported as an
easy, inexpensive, sustainable, and effective bio-sorbent for water de-
contamination (Purkayastha et al., 2014; Filote et al., 2017; Segovia-
Sandoval et al., 2018; Allahkarami and Rezai, 2019). Among these,
algae-based bio-sorbents that are renewable, cheap, available year-
round, and have high uptake capacity have gained attention for TM re-
moval applications (Yang et al., 2015; Amin and Chetpattananondh,
2019; Barquilha et al., 2019; Emparan et al., 2020). Once the bio-
sorption process is completed, algal biomass can be landfilled, con-
verted to bio-char or bio-fuel with a significantly lower impact on the
environment (Yu et al., 2017; Amin and Chetpattananondh, 2019; Nair
et al., 2019). Jaafari and Yaghmaeian (2019) indicated that Chlorella
Coloniales is an excellent, simple, and low-cost bio-sorbent that can re-
move up to 95% of As, Cd, Co, Cr, and Fe from wastewater. Moghazy
(2019) used Chlamydomonas for the removal of methylene blue (MB)
dye from aqueous solutions. The maximum removal efficiency of
80.8% was achieved with a bio-sorbent dose of 1.5 g/L. Sayadi and
Shekari (2017) reported an 85.6% removal of cadmium in 30 min
using 2 mg/L of Spirogyra at a pH of 6. Davis et al. (2003) showed that
the toxic metal removals by brown algae are double that of conventional
ion-exchange resins. Cheng et al. (2019) recommended further investi-
gation to understand the changes in the surface properties and the im-
provement in %TMrem after applying the chemical treatment on algae
strains.
The uptake and removals of TMs via bio-sorption processes were ex-
plained by different mechanisms such as micro-precipitation, chelation,
complexation, ion exchange, and physical adsorption (Barquilha et al.,
2019; Cheng et al., 2019). On the other hand, the bio-sorption of TMs
by algae was related to the electrostatic force between the negatively
charged functional groups on the surface of algae, such as polysaccha-
rides, proteins, amino, hydroxyl, and carboxyl groups and the positively
charged TMs (Sajadi et al., 2016; Sayadi et al., 2018). Accordingly, any
treatment process that can alter or improve the negative charge on
the surface algae would significantly enhance the %TMrem. Wang et al.
(2012) improved the adsorption capacity of rice stems by applying an
alkalinity treatment. The treated rice stems were effective at removing
trace uranium from an aqueous solution. Teixeira Tarley et al. (2004)
demonstrated that chemical treatment of rice husk using sodium hy-
droxide significantly improved the adsorption capacity of different
metals. He and Chen (2014) proved that the positively charged func-
tional groups on the surface of bio-sorbents play an essential role in
2
Science of the Total Environment 755 (2021) 142654
the removal of TMs. Luo et al. (2006) improved the TMs bio-sorption ca-
pacity of brown algae, Laminaria japonica, via chemical crosslinking
modification using epichlorohydrin potassium permanganate (PC).
Despite a large body of research on the use of algae for the removal
of TMs, most of the published works have focused on the removal of
TMs from synthetic wastewater under lab-controlled conditions. How-
ever, the effect of wastewater chemistry and the presence of different
contaminants on the bio-sorbent capacity and the overall %TMrem was
ignored. It is will-known that the water chemistry and the presence of
different contaminants (organic and inorganic) might compete with
targeted TMs for binding sites and decreasing the %TMrem (Acharya
et al., 2018). The removal of TMs from real wastewater by algae remains
a challenge due to the selectivity and complicity of the system, and the
undetermined optimum operational conditions that might limit the ap-
plication of this alternative at a scale (Moghazy, 2019). Besides, the fac-
tors affecting the improvement of the bio-sorption efficiency of algae
after a chemical treatment requires further investigation. Therefore,
the present study investigates for the first time the removal of TMs
from real industrial wastewater. The effect of acid chemical treatment
on improving the bio-sorption capacity of Spirulina platensis (SP) and
Chlorella vulgaris (CV) was explored are connected to the %TMrem. The
treated algae strains were used to remove aluminum (Al), nickel (Ni),
and copper (Cu) from wastewater, which were selected due to their
presence in different industrial effluents in Qatar. The effect of pH, bio-
sorption contact time (Ct), initial TM concentrations [TM]i concentra-
tion, and concentrations of bio-sorbent (Calgea) on the %TMrem, and the
optimal operational parameters were determined. In addition, the ther-
modynamics and mechanism of the bio-sorption process were
explored.
2. Material and methods
2.1. Chemicals
The wastewater (WW) used in tests are effluents of secondary
wastewater treatment plant (WWTP) from Doha industrial city, Qatar.
All the chemicals used in the present study were analytical grades pur-
chased from Sigma-Aldrich. The TM samples were prepared by adding
analytical grades of copper sulfate pentahydrate (CuSO4.5H2O, CAS #:
7758-99-8), nickel nitrate hexahydrate ((NiNO3)2.6H2O, CAS #:
10026-22-9) and aluminum sulfate (AL (SO4)3, CAS # 10043-01-3) sep-
arately to the WW. The pH of the WW was controlled using 0.1 N HNO3
or 0.1 N NaOH.
2.2. Cultivation and preparation of bio-sorbent
Spirulina platensis (SP, UTEX #1926, Texas, USA) and Chlorella
vulgaris (CV, UTEX # 265, Texas, USA) were grown in 3 L sterilized reac-
tors incubated at a temperature of 27° ± 0.5C. The light source to the re-
actors consisted of two fluorescent daylight lamps with an illuminance
of 3120 lx each. The photon flux intensity in the incubator was mea-
sured as 2.98 μW/cm2 using a radiometer (NIST Model IL 400A). Cultiva-
tion was performed by mixing 350 mL of growth medium with an initial
concentration of microalgae culture, on a dry base, of 2.4 × 10−3 gdw/L
and incubated for 21 days. The content and the preparation procedure
of the growth medium are presented in previous work (Almomani
et al., 2019b). Samples were mixed by swirling 8 times a day, CO2 was
supplied to the culture at 5 vol% and a flow rate of 1.35 mL/min. After
21 days, the algae were collected, washed 10 times with distilled
water (DW), dried overnight (24 h) at 100 °C, milled, subsequently
sieved with a 200-mesh size sieve and utilized in the experiments.
The chemical treatment of SP and CV was performed by mixing the pow-
der algae at a speed of 500 rpm with 0.5 M sulfuric acid at a ratio of
1:20 at 35 ± 1 °C. Thereafter, the algae were recovered by filtration
through a 0.45 μm filter (Whatman, USA), washed with DW until the
F. Almomani and R.R. Bhosale
pH of the water reached pH 7 ± 0.5, dried as before, and utilized in the
experiments.
2.3. Experimental procedure
The specific concentration of TMs was prepared by spiking the in-
dustrial wastewater with copper sulfate pentahydrate, nickel nitrate
hexahydrate, and aluminum sulfate. Equilibrium tests were carried
out by mixing specific doses (0.25, 0.5, 1, 1.5, 4 and 5 g/L) of dry bio-
sorbents (SP or CV) with 20 mL of wastewater containing specific con-
centrations (2.5–100 mg/L) of one of the TMs (Al, Ni, and Cu) at
150–200 rpm for 120 min. Once the test was completed, the algae
were separated from the wastewater and the concentration of TM in
the WW was measured. The concentration of TM in the WW was deter-
mined by testing filtrated samples in a flame atomic absorption system
(model AA-700). Quality assurance tests were performed to verify the
detection limit and showed that the detection limit can reach up to
1 ± 0.04 μg/L with the standard deviation (Std. Dev) ~ 0.089. The
amount of TM lost in all cases did not exceed 0.3%. The effect of contact
time on the %TMrem was carried out by repeating the previous proce-
dure at different time intervals of 5, 15, 20, 40, 60, 80, 100, or 120 min.
Similarly, other sets of tests were carried out to determine the effect
of pH in the range of 4–7, Calgae of 0.25, 0.5, 1, 1.5, 4, and 5 g/L, and
[TM]i of 2.5, 7.5, 15, 25, 75, and 100 mg/L on the %TMrem. All of the
tests were repeated three times and the average values at 95% confi-
dence are presented and discussed.
Zeta potential (ζ) was measured using a NanoBrook Zeta Potential
Analyzer (NanoBrook 90Plus Zeta, USA). Tests were performed by dilut-
ing the wastewater and algae in combination with Milli-Q water
(1:10 mL). Thereafter, a 1 mL of the mixture was added to the zeta po-
tential flow cell and the potential was recorded at 25 °C. A 1% HNO3 or
1 N NaOH were used to adjust the pH if required. The ζ test was repeated
at least three times for each sample and the results were reported as an
average value at 95% confidence level.
2.4. Mathematical manipulation
The percentage removal of toxic metal (%TMrem) was calculated
using Eq. (1)
C i;TM −C f ;TM
%toxic metal removal ð%TMrem Þ ¼  100% ð1Þ
C i ; TM
where Ci,TM and Cf,TM are the initial and final concentration of TM in the
wastewater (mg/L). The adsorption capacity of bio-sorption (i.e., algae)
was evaluated by the material balance on the system. The mass of TMs
adsorbed on the algae is equal to the difference between initial and final
concentrations in the WW. The mass of TM adsorbed on algae at equilib-
rium (qe, mg/mg) and at any time (qt, mg/mg) was calculated using
Eqs. (2) and (3)


C i;TM −C e;TM
qe ¼ ð2Þ
S


C i;TM −C t;TM
qt ¼ ð3Þ
S Fig. 1 presents the surface properties and surface functional groups
where Ct,TM is the concentration of TM in the WW (mg/L) at a specific
time. S is the mass of algae (mg) per volume of WW (L). The bio-
sorption of TMs on algae was tested against Langmuir (Eq. (4)) and
Freundlich (Eq. (5)) isotherms.
qmax K L C e;TM
qe ¼ ð4Þ
1 þ K L Ce;TM

1
qe ¼ K f C e;TM n ð5Þ
3
Science of the Total Environment 755 (2021) 142654
where qmax is the maximum TM adsorbed on a unit mass of algae (mg/
mg), Kf is the adsorption capacity of algae and, n is the adsorption inten-
sity The changes in free energy during the bio-sorption process were
calculated using the apparent Gibbs free energy (Δ G°) Eq. (6).
ΔG ° ¼ −RTln K L ð6Þ
where T is the temperature (K), R is the ideal gas constant (8.314 J.
Mol−1.K−1), and KL is the equilibrium constant calculated as (qe/ce).
The TMs bio-sorption kinetics was followed using pseudo-first, Eq. (7),
and second-order, Eq. (8)
Lnðqe −qt Þ ¼ lnqe −k1 t ð7Þ
t 1 t
¼ þ ð8Þ
qt k2 q2e qe
2.5. Optimization of TM removals
The optimum operational conditions for %TMrem was determined
using response surface methodology (RSM). The RSM combines a math-
ematical and statistical approach to identify the significance of each op-
erating factor on %TMrem. The effect of [TM]i, Ct, pH, and the
concentrations of bio-sorbent (Calgae) were tested for the maximum %
TMrem. The RSM was identified based on a central level between the
lowest and the highest responses. Experimental results at differing Ct
(min) of 5, 15, 20, 40, 60, 80, 100 and 120 min., a pH of 3, 4, 5, 6, and
7, and Calgae, of 0.25, 0.5, 1, 1.5, 2, and 2.5 g/L were used in the calcula-
tions. The %TMrem was fitted into a quadratic model (Eq. (9)) and the
significance of the model was evaluated using the ANOVA statistical
method using Prism Software V 7.04.
X
k X
k X
k−1 X
k
%TMrem ¼ B0 þ Bi Z i þ Bii X 2i Bij X i X j ð9Þ
i¼1 i¼1 i¼1 j¼2
where Zi, Zj, …., Zk are input operational parameters ([TM]i, Ct, pH, and
Calgae), B0 is the intercept, Bi (i = 1, 2, …, k) is the linear effect, Bii (i = 1,
2, …, k) is the squared effect, and Bij (i = 1, 2, …, k-1, j=, 2, …,k) is the
interaction effect. The root means square (RMS) and correlation
coefficient.
2.6. Statistical analysis
The statistical significance of the reported %TMrem at different oper-
ational conditions was judged using a one-way analysis of variance
(ANOVA). Prism GraphPad statistics software (Version 7.04, USA) was
used to conduct the statistical calculation at a 95% confidence interval
(5% confidence level).
3. Results and discussion
3.1. Characteristics of treated and untreated algae
of SP and CV before and after the acidic treatment (Ac-T) with H2SO4.
The structure of SP has a peak at 3435 1/cm correspondings to −OH
functional group, a peak at 2922 corresponding to −CH group, and pow-
erful bands in the range of 1544–1630–1659 1/cm corresponding to am-
ides (Fig. 1a). The observed vibration bans at 1030 and 1409 1/cm were
related to the presence of \\S\\O and \\P\\O respectively. After the
acid treatment, peaks in the range of 1355–1165 1/cm were appeared
and were subsequently identified as S_O. Other peaks at 852, 755,
545 1/cm were observed and associated with N\\H, \\C\\H, and
O\\P\\O groups, respectively. There are no previous results outlined
in the relevant literature to compare these results with, and currently,
F. Almomani and R.R. Bhosale Science of the Total Environment 755 (2021) 142654

SP (a) 3.2. TM removal by SP and CV

(treated)
Fig. 2 illustrates a significant improvement in the percentage of toxic
metals (Al, Ni, and Cu) removal (%TMrem) by both algae strains follow-
ing the Ac-T. The %TMrem was 1.8 and 1.5 folds higher for the treated
%transmittance

SP and CV in comparison with untreated strains. While the %TMrem by

S=O
the treated SP reached up to 95.0 ± 0.3% (Std. Dev = 0.24), 87.0 ±
(Untreated) 0.2% (Std. Dev = 0.34), and 62.0 ± 0.3% (Std. Dev = 0.50) of Ni, Al,
and Cu, the untreated SP removed only 69.0 ± 0.3% (Std. Dev = 0.05),
45 ± 0.4% (Std. Dev = 0.08), and 61 ± 0.4% (Std. Dev = 0.02), respec-

O-P-O
N-H
C-H
tively. The %TMrem using CV was 87.0% ± 0.2 (Std. Dev = 0.09), 79.1 ±

C-H
0.4% (Std. Dev = 0.08), and 80.0 ± 0.2% (Std. Dev = 0.04) for treated,
and 52 ± 0.2% (Std. Dev = 0.04), 49 ± 0.5% (Std. Dev = 0.07), and

S=O
N-H 52 ± 0.2% (Std. Dev = 0.10) for the untreated strain, respectively. The
OH

noticeable improvement in the %TMrem after Ac-T was related to the in-
C=C
C-H

crease in the portion of the negatively charged functional groups on the

surface of algae. As the FTIR results indicate, the Ac-T increased the in-
tensity of the ionic groups (−OH, N\\H and O\\P\\O), which have a
4000 3500 3000 2500 2000 1500 1000 500
stronger electro-attraction force to the cationic TMs the conditions
Wavelength (1/cm) that increase the reported %TMrem. Conversely, untreated SP and CV
contain less negatively charged functional groups on their surface,
with a weak attachment to cation, which results in a lower %TMrem.
The obtained results revealed that the studied metals exhibited dif-
CV (Treated) (b) ferent %TMrem with each strain. In general, SP achieved higher %TMrem
compared with CV for all TMs (Al, Ni, and Cu). Likely, the high intensity
of negative charge on the surface of SP with higher attraction force to
TMs is the main reason for the observed higher removal efficiency.
Moreover, as Ni has the highest cationic (positive charge) properties
%transmittance

followed by Cu, and then A1 it is logical to attain the highest %TMrem.

S=O

As the untreated algae strains exhibited a lower %TMrem, much of the

S=O

following discussion focuses on treated SP and CV strains, and their ca-

S=O

pability for the removal of TMs.

(Untreated)
3.3. Effect of pH on %TMrem

Fig. 3 presents the effect of pH, in the range of 3–7 on the percentage
O-P-O
C-H

C-O

removal of Al, Ni, and Cu with treated and untreated SP and CV. Tests
O=H

were conducted at 25 ± 0.5 °C, with [TM]i of 50.0 ± 0.2 mg/L (Std.
N-H

C=C

4000 3500 3000 2500 2000 1500 1000 500
Wavelength (1/cm)
Fig. 1. FTIR spectrum of treated and untreated algae strains (a) SP and (b) CV.
the only reference available is the work of Sheng (2010) who
highlighted similar observations. Additionally, the modified SP
(Fig. 1a) exhibited a shift and an increase in the intensity for the peaks
in the range of 4000–400 1/cm. This suggests that the functional groups
on the SP surface were changed to more electronegative functional
groups, which is expected to increase the bio-sorption of TMs. Fig. 1b
exemplifies the Fourier-transform infrared spectroscopy (FTIR) of CV
before and after the chemical treatment. The peaks at 3443, 2070, and
1654 1/cm in the untreated CV were related to NH group, N_C_S
group, and type I and II of amide. The treated CV displayed a new peak
at 1387 1/cm corresponding to −OH group. Other peaks at 1037, 1155,
and 1345 1/cm indicate the presence of S_O. An additional peak at
1076 1/cm was related to C_C and C_O groups. The peak at 580 1/
cm confirms the presence of O-P-O. Ferreira et al. (2011) observed a
similar functional group on the surface of CV during the study of bio-
sorption of Ni2+, Zn2+ and Pb2+. Both strains obtained higher portions
of negative functional groups, which suggests that the Ac-T developed a
greater electronegative surface than the original strain.
Dev = 0.54) and Ct of 120.0 ± 0.5 min (Std. Dev = 0.14). The %TMrem
is highly dependent upon the pH of the wastewater, with a general
trend exemplifying lower removal efficiency at an acidic pH. The lowest
%TMrem for the three metals, using treated and untreated algae strains
occurred at pH = 3. Increasing the pH exhibited an increase in the
rate of adsorption and %TMrem until the optimal pH was reached; there-
after, the %TMrem decreased. The results indicated that treated SP
achieved a maximum percentage Al removal of 83.7 ± 0.2% (Std.
Dev = 0.04), at pH = 6. However, Ni and Cu had a maximum percent-
age removals of 82.0 ± 0.5% (Std. Dev = 0.08) and 80.0 ± 0.4% (Std.
Dev = 0.07) at pH = 5.0 and 6, respectively. Untreated SP exhibited
the maximum %TMrem of 63.0 ± 0.3% (Std. Dev = 0.14), 54.1 ± 0.3%
(Std. Dev = 0.54), and 40.0 ± 0.3% (Std. Dev = 0.24) at a pH of 4, 6,
and 7 for Ni, Al and Cu, respectively. Lower %TMrem were observed by
treated CV, with the maximum removals of 87.1 ± 0.2% (Std. Dev =
0.23), 78.2 ± 0.2% (Std. Dev = 0.08), and 78.0 ± 0.4% (Std. Dev =
0.24) for Ni, Al, and Cu at a pH of 5, 6 and 7, respectively. Untreated
CV exemplified the maximum percentage Ni, Al, and Cu removals of
54.3 ± 0.2% (Std. Dev = 0.77), 47.3 ± 0.2% (Std. Dev = 0.01), and
49.0 ± 0.2% (Std. Dev = 0.23) at a pH of 5, 6 and 7, respectively. As
discussed before, the improvement in the %TMrem after the Ac-T may
be connected to the formation of negatively charged surface functional
groups on the surface of algae with high attraction force to the TMs,
which are the conditions that improve the metal-algae attachment
and increase their removals. The change in the %TMrem as a function of
pH may be associated to the competition effect between H+ and TMs
to adsorb on the surface of algae. At a low pH, the water solution con-
tains a high concentration of H+, which competes to negative charge

4
F. Almomani and R.R. Bhosale Science of the Total Environment 755 (2021) 142654

100 (a) 100 (b)

80 80

%TMrem

%TMrem
60 60

40 Ni 40

Al Ni
20 20 Al
Cu Cu
0 0
0 20 40 60 80 100 120 140 0 20 40 60 80 100 120 140
Time (min) Time (min)

100 (c) 100 Ni (d)

Al
80 80 Cu
%TMrem

%TMrem
60 60

40 40
Ni
20 Al 20
Cu
0 0
0 20 40 60 80 100 120 140 0 20 40 60 80 100 120 140
Time (min) Time (min)

Fig. 2. %TMrem by treated and untreated algae strains as a function of time (a) treated SP, (b) untreated SP, (c) treated CV, and (d) untreated CV. Conditions: CAlgae = 2.5 ± 0.1 g/L
(Std. Dev = 0.14), T = 25 ± 0.5 °C, pH = 6 and [TM]i = 75.0 ± 0.3 mg/L (Std. Dev = 0.45).

100 80
treated SP (a) Untreated SP (b)

80 60
%TMrem

%TMrem

40
60

20
40
0
2 3 4 5 6 7 8 2 3 4 5 6 7 8
PH PH
100 100
Treated CV (c) Untreated CV (d)
80 80
%TMrem
%TMrem

60 60

40 40

20 20

0 0
2 3 4 5 6 7 8 2 3 4 5 6 7 8
PH PH
Al Ni Cu

Fig. 3. Effect of pH on the %TMrem using treated and untreated SP and CV. Conditions: CAlgae = 2.5 ± 0.1 g/L (Std. Dev = 0.06), T = 25 ± 0.5 °C, [TM]i = 50.0 ± 0.2 mg/L (Std. Dev = 0.54)
and Ct = 120.0 ± 0.5 min (Std. Dev = 0.14).

5
F. Almomani and R.R. Bhosale
with cation metals (Ni, Cu and Al) and decreases the %TMrem. Results
further indicated that H+ might reduce the negative surface charge on
the algae surface, while increasing the repulsion force, leading to a sub-
stantial decrease in the reported %TMrem. Moreover, under acidic condi-
tions, the solubility of metals in water increases, which results in a
decrease in the TM sorption rates. Increasing the pH of the wastewater
correspondingly decreases the solubility of TMs, reduces the concentra-
tion of H+, and increases the concentration of OH−. Under these condi-
tions, the solubility of TMs is low and all of the negatively charged
functional groups on the surface of algae are available for TM adsorp-
tion. Thus, a high %TMrem was observed. For the treated SP, the maxi-
mum %TMrem were obtained at a pH of 6 for A1, 5 for Ni, and 6 for Cu.
Alternatively, the maximum %TMrem(s) for the treated CV were achieved
at 6 for A1, 5 for Ni, and 7 for Cu. The differences in the required pH to
attain a maximum %TMrem may be correlated to the variation in the
TM cationic properties (Ni > Cu > Al). Ni has the highest cation proper-
ties with a required pH value around 5, followed by Cu and Al with the
required pH > 6.
The dependence of the %TMrem on the corresponding pH may be cor-
related to the surface functional groups of algae, the impact on the me-
tallic ions, and the surrounding water matrix. The Ac-T modified the
surface functional groups of algae, increased the ionic ligands of the neg-
atively charged functional groups (e.g. carboxyl, phosphate, and amino
groups), enhanced the chance for algae-TM binding via electrostatic ad-
sorption, and ultimately increased the reported %TMrem Sayadi et al.
(2018); He and Chen (2014). Sajadi et al. (2016) and Sayadi et al.
(2018) demonstrated that the presence of the negatively charged func-
tional groups, such as the carboxyl group, increased the attraction force
between algae-TM and improved the %TMrem. As the pH decreases, the
surface charge of algae changes to positive, which reduces the algae-TM
attraction force, and also decreases the %TMrem as confirmed by He and
Chen (2014). Additionally, the variations in the pH directly affect the
chemistry of the TMs and the natural organic matters (NOMs) in the
wastewater. Increasing the pH above 7 may simultaneously increase
the chance for TMs precipitation, which would decrease the mass of
adsorbed metal and reduce the %TMrem (Sajadi et al., 2016). However,
acidic pH increases the solubility of TMs and may reduce the %TMrem
as well. Additionally, the surrounding water matrix due to the presence
of NOMs contributes to the %TMrem. The presence of NOM in wastewa-
ter will likely affect the distribution of algae and TMs, as well as the cor-
responding surface charge. Furthermore, the NOM might compete with
the TMs on the algae active site and reduce the reported %TMrem. The
TMs removal mechanism as a function of the pH in actual wastewater
was verified by measuring the zeta potential (ζ) of the TM solution
and the treated algae strains. Results indicated that the ζ values of the
treated SP and CV change as a function of pH to become more negative,
with a pH in the range of 5–6 for the SP, and 6–7 for the CV. The ζ values
of the studied TMs solutions are positive under all of the pH conditions.
However, there was a direct correlation between the ζ value of TMs and
the concentration of NOM. Increasing the concentration of NOM in the
wastewater exemplified a decrease in the ζ toward negative values.
The electrostatic attraction force between the negatively charged algae
and the positively charged TMs establishes the high algae-TM attraction
force, as well as promotes greater %TMrem. The values of ζ for SP and CV
are < −28 mV for tests carried out with a pH < 4 or a pH > 7. This sug-
gests stable algae in suspension that cannot coagulate with TMs due to
high repulsion forces. Tests carried out with pH values in the range of
5–7 exhibited ζ values > −18 and −20 mV for SP and CV, respectively.
This indicates that there is a high tendency of the algae strains to coag-
ulate with TMs, which helps to achieve a higher %TMrem. The observed
results illustrate how accurate ζ values are important for predicting
and controlling the stability of algae-TM aggregates. Algae strains with
a ζ value (>25 mV, positive or negative) indicate that the particles are
more likely to remain in suspension due to strong repulsing forces.
However, as the ζ value decreased the chance for coagulation and the
%TMrem increases. Vandamme et al. (2013) demonstrated that the
6
Science of the Total Environment 755 (2021) 142654
value of ζ is dependent upon the pH and ionic strength (IS) of wastewa-
ter, which impacts the width of the electric double layer (EDL). Increas-
ing the IS of the wastewater led to a decrease in the electric double layer
width. Thus, the resulting interaction (attractive/repulsive) is depen-
dent upon the character of the charge of interacting surfaces and on
the properties of the surrounding wastewater. Miao et al. (2015) dem-
onstrated that the surface charge of CuO-nanoparticles exhibited a pos-
itive ζ at low concentrations of NOM (<1 mg/L), and above that, the ζ
became negative. This effect suggests that as the concentration of
NOM increases, the ζ became gradually more negative, and decreased
the attraction force between TMs and algae, hindering the bio-
sorption and decreasing the %TMrem.
3.4. The effect of contact time (Ct)
Fig. 2 presents the effect of contact time (Ct) on the %TMrem (A1, Ni,
and Cu) with treated and untreated algae strains. As the Ct increased,
the %TMrem progressively increased due to the corresponding increase
in the change of TM-algae interaction, which obtained equilibrium
after 80 ± 2 min (Std. Dev = 0.56) in most cases. Both strains demon-
strated a fast TM removal rate during the first 20 ± 2 min and thereafter
slowed down. The effect of the chemical treatment is clear, given the
high removal rate at all of the tested times in comparison to the un-
treated algae. Ni exhibited the highest removal rate, followed by Cu,
and then A1. The observed high adsorption rate occurring at the initial
time is due to the high availability of active sites and the high concentra-
tion of TMs, which increases the adsorption driving force. Besides, it was
observed that the adsorption kinetics of A1, Ni, and Cu via the treated SP
and CV were fast. The equilibrium concentration of Ni was achieved
after 40 ± 1 and 60 ± 1 min for treated SP and CV, respectively. The %
TMrem with the treated SP after 40 ± 1 min was 92.5 ± 0.2% (Std.
Dev = 0.34), 800 ± 0.2% (Std. Dev = 0.04), and 63 ± 0.3% (Std.
Dev = 0.14) for Ni, Cu and Al, respectively; for the treated CV, the %
TMrem removal was 63.5 ± 0.2% (Std. Dev = 0.13), 63.1 ± 0.2% (Std.
Dev = 0.13), and 62.5 ± 0.1% (Std. Dev = 0.10) for Ni, Cu, and A1, re-
spectively. After 80 min, the %TMrem increased to 93.1 ± 0.3% (Std.
Dev = 0.22), 91.5 ± 0.3% (Std. Dev = 0.22), and 85.1 ± 0.3% (Std.
Dev = 0.22) with SP, and 83.5 ± 0.5% (Std. Dev = 0.15), 83.1 ± 0.5%
(Std. Dev = 0.15), and 79.5 ± 0.5% (Std. Dev = 0.15) with CV,
respectively.
3.5. Effect of the concentrations of bio-sorbent (Calgae)
Fig. 4 illustrates the effect of the concentrations of bio-sorbent
(Calgae), in the range of 0.25, 0.5, 0.75, 1.0, 1.5, 2 and 2.5 mg/L, on the %
TMrem using treated and untreated SP and CV. Increasing the Calgae of
the treated and untreated algae increased the % TMrem of Al, Ni and Cu
due to the increase in the available active sites for TM uptake. Thereaf-
ter, the increase in Calgae doses did not exhibit any discernible changes
in the %TMrem. The limited effect of the high concentration of the Calgae
on the %TMrem is likely due to the agglomeration of algae strains that re-
duce the available active sites for bio-sorption. This was verified by
measuring the ζ values of CV and SP under high bio-sorbent doses. In-
creasing the concentration of SP and CV was combined with an increase
in the ζ to >−26 and −25 mV for SP and CV, respectively. Nimesh et al.
(2017) suggested that particles with ζ ≥ −25 mV would gradually ag-
glomerate due to inter-particle interactions, such as van der Waals
and hydrophobic forces. Therefore, it was noticed that as the Calgae in-
creased from 0.25 ± 0.03 g/L to 2.5 ± 0.03 g/L, the %TMrem increased.
The reported % TMrem 0.35 ± 0.03 g/L of treated SP removed 39.1 ±
0.2% (Std. Dev = 0.34), 40.0 ± 0.3% (Std. Dev = 0.50), and 28.1 ±
0.3% (Std. Dev = 0.24) of Al, Ni, and Cu, respectively; while the treated
CV exemplified 36.1 ± 0.2 (Std. Dev = 0.09), 39.1 ± 0.4% (Std. Dev =
0.08), and 34.1 ± 0.4% (Std. Dev = 0.08) removal at identical doses, re-
spectively. Increasing the bio-sorbent dose to 2.50 ± 0.03 g/L removed
89.1 ± 0.3% (Std. Dev = 0.50), 94.1 ± 0.3% (Std. Dev = 0.24), and
F. Almomani and R.R. Bhosale Science of the Total Environment 755 (2021) 142654

100 treated SP (a) 100 Untreated SP (b)

80 80

%TMrem

%TMrem
60 60

40 40
AL AL
20 20
Ni Ni
Cu Cu
0 0
0 1 2 3 4 5 6 0 1 2 3 4 5 6
Calgae (g/L) Calgae (g/L)

100 Treated CV (c) 100 Untreated CV (d)

80 80
%TMrem

%TMrem
60 60

40 40
Al
20 20
Al
Ni Ni
Cu Cu
0 0
0 1 2 3 4 5 6 0 1 2 3 4 5 6
Calgae (g/L) Calgae (g/L)

Fig. 4. %TMrem as a function of algae concentration (Calgae) for treated and untreated SP and CV. Conditions: [TM]i = 55.0 ± 0.2 mg/L (Std. Dev = 0.54), T = 25 ± 0.5 °C, pH = 6, with a
contact time of 120 min.

66.0 ± 0.3% (Std. Dev = 0.24) of Al, Ni and Cu for treated SP, and 79.10.2
(Std. Dev = 0.09), 90.1 ± 0.4% (Std. Dev = 0.08), and 82.0 ± 0.4% (Std.
Dev = 0.08) for treated CV, respectively. Fomina and Gadd (2014)
showed that the %TMrem increased by simultaneously increasing the ad-
sorbent dosage to a certain limit, though there was a gradual decrease
with the corresponding increase in the initial metal concentration due
to absorbed cations filling up the active sites. Lee and Chang (2011)
demonstrated that TM adsorption is indirectly proportional to biomass
concentration. As such, the adsorption rate reduces by increasing the
adsorbent dose. Whereas, the work of Bhatti et al. (2010) highlighted
that the increase in the bio-sorbent dosage led to a decrease in the re-
ported %TMrem. It was also suggested that the interference of bio-
sorption sites with one another resulted in intracellular electrostatic in-
teraction forces and reduced the %TMrem. The attraction between algae-
TMs was examined by FTIR, as such, the metallic ions were observed to
be attached to the peaks related to OH, N\\H, and S_O. The obtained
results confirmed that TMs were successfully attached to the SP and
CV treated surface via electrostatic force after the ion exchange of H+
with water. Moreover, the results confirmed that the algae-TM bio-
sorption mechanism is governed by the complexation of these func-
tional groups on the surface of algae with the TMs (El-Boubbou et al.,
2007; Shurair et al., 2019; Almomani et al., 2019c).
3.6. Effect of initial concentration of metals
An additional set of experiments was carried out to explore the effect
of the initial concentration of TM ([TM]i) on the %TMrem with treated
and untreated SP and CV. Increasing the [TM]i from 50 to 75 mg/L de-
creased the %TMrem by 6.0 ± 0.2% (Std. Dev = 0.54) to 8.5 ± 0.2%
(Std. Dev = 0.54) and from 7 ± 0.3% (Std. Dev = 0.33) to 9 ± 0.3%
(Std. Dev = 0.33) for SP and CV, respectively. The observed trends indi-
cate that filling of the active sites on algae surface reduces the
adsorption capacity, and decreased the %TMrem. Furthermore, it is likely
that the diffusion of the TMs through the SP and CV active sites is pref-
erable at lower [TM]i. The results presented throughout this study are
consistent with the results obtained by Mirghaffari et al. (2015) who
showed that the %TMrem of cadmium (Cd) and lead (Pb) by green
microalgae decreased by increasing the [TM]i due to limited internal
adsorption.
3.7. Adsorption isotherm
Freundlich and Langmuir's isotherms were tested against the exper-
imental data. Table 1 displays the results of the regression analysis of the
isotherms with the corresponding R2. The removal of TMs with treated
SP and CV follows Langmuir and Freundlich isotherms. The regression
analysis of the data showed R2 ≥ 92. The qmax of Langmuir models indi-
cated that TM attachment to algae occurs through mono-layer diffusion,
which is highest for Ni with qmax in the range of 49.32 ± 0.05 to 21.3 ±
0.05 mg/mg for treated SP, and in the range of 43.89 ± 0.04 to 18.96 ±
0.03 mg/mg for the treated CV. Aluminum (Al) had the second-highest
qmax in the range of 47.8 ± 0.03 to 15.21 ± 0.03 mg/mg for treated SP,
and 42.54 ± 0.04 to 13.54 ± 0.04 mg/mg for the treated CV. The lowest
qmax was observed for Cu at 38.90 ± 0.02 to 15.01 ± 0.02 mg/mg for
treated SP, and 39.10 ± 0.03 to 12.54 ± 0.03 mg/mg for the treated
CV. The values of KL are high for Ni, which ranged from 0.32 ± 0.02 to
0.11 ± 0.02 L/mg for the treated SP, and from 0.27 ± 0.03 to 11.00 ±
0.05 L/mg for the treated CV. The values for Al and Cu are 85–77% less
for treated SP, and 82–74% less for treated CV. The TMs removed with
treated SP and CV are well described by the Freundlich model. This sug-
gests that the bio-sorption of TMs onto these strains occurs via electro-
static forces due to the development of the attraction layer between the
TMs and algae surface. The magnitude of Kf and n describe the required
energy for TM bio-sorption on algae strain(s). The bio-sorption easily

7
F. Almomani and R.R. Bhosale Science of the Total Environment 755 (2021) 142654

Table 1
Langmuir and Freundlich isotherms parameters and Gibbs free energies for treated SP and CV.

Isotherm Dose Al Ni Cu
(mg/L)
qmax KL R2 ΔG° qmax KL R2 ΔG° qmax KL R2 ΔG°
(mg/mg) (L/mg) (kJ/mol) (mg/mg) (L/mg) (kJ/mol) (mg/mg) (L/mg) (kJ/mol)

Langmuir SP
0.25 41.0 ± 0.02 0.23 ± 0.03 0.90 −1.133 42.55 ± 0.07 0.32 ± 0.02 0.87 −1.23 38.90 ± 0.02 0.65 ± 0.08 0.93 −1.33
0.50 47.8 ± 0.03 0.25 ± 0.05 0.92 −1.043 49.32 ± 0.05 0.28 ± 0.06 0.92 −1.18 37.3 ± 0.04 0.23 ± 0.06 0.93 −1.16
1.00 23.3 ± 0.05 0.14 ± 0.07 0.94 −0.90 27.31 ± 0.04 0.18 ± 0.05 0.87 −1.09 20.2 ± 0.06 0.26 ± 0.03 0.92 −1.05
1.5 15.21 ± 0.04 0.09 ± 0.03 0.84 −0.88 21.3 ± 0.05 0.11 ± 0.04 0.89 −0.96 15.0 ± 0.02 0.20 ± 0.06 0.94 −1.01
CV
0.25 34.51 ± 0.03 0.22 ± 0.07 0.93 −1.02 37.87 ± 0.04 0.27 ± 0.03 0.90 −1.13 34.62 ± 0.05 0.62 ± 0.07 0.96 −1.20
0.50 42.54 ± 0.04 0.24 ± 0.04 0.95 −0.94 43.89 ± 0.04 0.27 ± 0.02 0.95 −1.10 39.10 ± 0.04 0.22 ± 0.06 0.96 −1.04
1.00 20.74 ± 0.05 0.13 ± 0.01 0.97 −0.81 24.31 ± 0.01 0.17 ± 0.04 0.90 −1.02 26.88 ± 0.04 0.25 ± 0.08 0.93 −0.95
1.50 13.54 ± 0.04 0.086 ± 0.05 0.87 −0.77 18.96 ± 0.03 0.11 ± 0.04 0.92 −1.03 12.54 ± 0.03 0.19 ± 0.03 0.97 −0.90

Isotherm Dose n Kf R2 n Kf R2 n Kf R2
(mg/L) (mg/mg) (mg/mg) (mg/mg)

Freundlich SP
0.30 2.4 ± 0.1 17.1 ± 0.2 1.00 2.7 13.1 ± 0.2 1.00 2.6 13.4 ± 0.4 0.98
0.50 2.3 ± 0.2 12.4 ± 0.1 0.94 2.4 12.5 ± 0.1 1.00 2.4 10.5 ± 0.3 0.95
1.0 1.9 ± 0.3 8.6 ± 0.2 1.00 2.2 7.8 ± 0.4 1.00 1.8 9.7 ± 0.6 0.94
1.50 1.9 ± 0.4 6.5 ± 0.1 0.95 1.7 5.6 ± 0.3 1.00 1.5 6.5 ± 0.3 0.92
CV
0.30 2.1 ± 0.3 16.2 ± 0.3 1.00 2.4 12.4 ± 0.6 1.00 2.3 12.7 ± 0.3 0.97
0.50 2.0 ± 0.2 11.8 ± 0.4 1.00 2.1 11.9 ± 0.3 1.00 2.1 10.0 ± 0.2 0.94
1.00 1.7 ± 0.4 8.2 ± 0.3 1.00 2.0 7.4 ± 0.4 0.90 1.6 9.2 ± 0.3 0.93
1.50 1.6 ± 0.2 6.2 ± 0.4 1.00 1.5 5.3 ± 0.5 1.00 1.3 6.2 ± 0.3 0.90

occurs when the 1/n value is between 0.1 and 1, and it is difficult to
adsorb if 1/n value is above 1 (Erentürk and Malkoç, 2007; Wu and
Yu, 2007). As presented in Table 1, the 1/n value for all of the studied
TMs is less the 1, confirming that the bio-sorption of TMs is favorable.
The Gibbs free energy (Δ G°) calculations showed that the TMs re-
moval by algae is spontaneous with Δ G° values in the range of
−1.133 to −0.900 for Al, −1.23 to 01.09 for Ni, and −1.33 to
−1.05 using treated SP and CV. Untreated SP and CV exhibited
90–86% lower Δ G° values.
3.8. Kinetics models
The experimental results were tested against the pseudo-first and
second-order kinetic models (Eqs. (7) and (8)). Regression analysis
showed that bio-sorption of TMs on algae fits the second-order kinetic
model with R2 of 0.95, 0.96, and 0.98 for Ni, A1, and Cu bio-sorption
on the treated SP, and 0.98, 0.93, and 0.97 for the aforementioned metals
on the treated CV. The kinetic constants were found to be 0.005 ±
0.0003, 0.004 ± 0.0002, and 0.004 ± 0.0002 g.mg/min for treated SP,
and 0.004 ± 0.0003 and 0.003 ± 0.0002 for treated CV, respectively.
The calculated kinetic constants are consistent with the values reported
by Abbas et al. (2014) and Farooghi et al. (2018).
3.9. Optimized TM removal efficiency
The %TMrem of Al, Ni, and Cu on the treated SP and CV were opti-
mized using the response surface methodology-statistical approach
(RSM-SA). The experimental results were successfully fitted with
R2 ≥ 0.88 into the second-order Eq. (10), corresponding to four indepen-
dent factors, including [TM]i (mg/L), Ct (min), pH, and Calgae (g/L).
%HM ¼ B0 þ B1 ½TMi þ B2 Ct þ B3 pH þ B4 CAlgae þ B5 ½TMi  Ct
þ B6 ½TMi  pH þ B7 ½TMi  CAlgae þ B8 Ct  pH þ B9 Ct

2
 C Algae þ B10 pH  C Algae þ B11 ½TMi þ B12 ðCt Þ2

2
þ B13 CAlgae þ B14 ðpHÞ2 ð10Þ
Eq. (10) was manipulated using ANOVA statistical software at a con-
fidence level of 95% (α = 5%). Insignificant terms with p > 0.05 were ig-
nored and only terms with p < 0.05 were accepted. Table 2 presents the
significant parameters for predicting the %TMrem by treated SP and CV.
This was validated using the statistical student's test (t-test) at α =
5%. The statistical manipulation showed individual parameter ([TM]i,
Ct, pH, and Calgae) has a significant contribution to the %TMrem by
algae, while the combined effect of these parameters are insignificant,
except for Ct ∗ CAlgae. The negative coefficient terms in the model suggest

Table 2
ANOVA results of the process parameters by response surface methodology-statistical approach (RSM-SA).

Parameter SP CV

Al Ni Cu Al Ni Cu

B0 93.50 ± 0.7(0.85)a 62.50 ± 0.4(0.05) 87.50 ± 0.8(0.65) 92.50 ± 0.6(0.45) 60.50 ± 0.6(1.25) 89.30 ± 0.6(1.25)
B1 −2.50 ± 0.08(0.75) 4.30 ± 0.03(0.15) 6.20 ± 0.04(0.95) −2.00 ± 0.02(1.65) 4.13 ± 0.02(0.15) 5.83 ± 0.06(0.89)
B2 0.96 ± 0.07(1.85) −0.13 ± 0.01(1.22) −0.13 ± 0.02(1.25) 0.77 ± 0.01(2.45) −0.12 ± 0.01(2.05) −0.12 ± 0.01(0.85)
B3 −1.92 ± 0.01(1.35) 0.39 ± 0.02 (1.65) 0.39 ± 0.04(2.44) −1.54 ± 0.06(2.45) 0.37 ± 0.08 (7.85) 0.37 ± 0.06(0.25)
B4 0.60 ± 0.05(1.65) 0.07 ± 0.03(0.95) 0.07 ± 0.01(3.10) 0.48 ± 0.04(2.22) 0.07 ± 0.01(2.15) 0.06 ± 0.01(0.25)
B9 −0.01 ± 0.005(2.85) −0.83 ± 0.001 (0.16) −0.10 ± 0.004(2.05) −0.08 ± 0.05(3.22) −0.80 ± 0.05(1.00) −0.09 ± 0.02(2.30)
B11 0.04 ± 0.008(3.15) 0.07 ± 0.007(0.23) −0.06 ± 0.001(1.55) 0.03 ± 0.01(1.62) 0.07 ± 0.01(2.55) −0.06 ± 0.04(1.45)
B12 −0.88 ± 0.02(5.85) −0.83 ± 0.03(0.11) −0.77 ± 0.03 (2.15) −0.70 ± 0.04(2.17) −0.80 ± 0.01(1.65) −0.72 ± 0.03(2.25)
B13 −0.80 ± 0.03(1.45) −0.90 ± 0.04(0.35) −0.66 ± 0.02(1.85) −0.64 ± 0.01(1.42) −0.86 ± 0.02(0.67) −0.62 ± 0.09(3.25)
B14 −1.28 ± 0.07(0.25) −0.01 ± 0.003(0.25) −0.02 ± 0.004(1.45) −1.02 ± 0.02(0.42) −0.01 ± 0.007(1.77) −0.02 ± 0.009(2.25)
a
Standard deviation.

8
F. Almomani and R.R. Bhosale Science of the Total Environment 755 (2021) 142654

the presence of optimum values to achieve the maximum %TMrem.
Fig. 5a presents the response surface of the %TMrem as a function of Calgae
and pH at constant [TM]i of 75 mg/L, while Fig. 5b shows the effect of Ct
and Calgae at a constant pH of 7. The model exemplifies that the %TMrem
can be increased by increasing Ct and CAlgae and, pH up to specified
values (90 min, 4.9 g/L and 6.6, respectively). Thereafter, the %TMrem de-
creased. The maximum percentage removal of toxic metals %TMrem, max
was determined by finding the local maximum from the developed
trend. This was determined to be at a pH = 6, CAlgae of 4.6 ± 0.2 (g/L,
and Ct of 87 ± 1 min for the SP and pH = 6.5, CAlgae of 4.9 ± 0.1 g/L,
and Ct of 89 ± 1 min for the CV. The RSM results confirmed that the %
TMrem is dependent on the pH with lower removal efficiency achieved
at acidic pH, which was related as discussed before to the competition
adsorption between H+ and TMs on the surface of algae. The increase
in the TM-Algae contact time has also a positive contribution to the pro-
gressive increase in the %TMrem. However, this parameter should be
carefully determined as high Ct did not show a significant improvement
in the %TMrem and might result in high reactor volume at large scale ap-
plications. Other parameters (Calgae and [TM]i) would be determined
based on real treatment conditions, with the first parameter depends
100
(a)
90
80
%HM 70
on the available algae strains and the corresponding growth conditions,
while the second is related to the industrial wastewater characteristics
that depend on the real-life industrial activities. Given the results, the
removal of TMs using algae can be employed within the wastewater
treatment process to complete the removal of any remaining traces of
metal. It is proposed to install this treatment unit after the secondary
treatment to benefit from the nutrients and organic matter that remains
in the wastewater for algae growth. This is immensely helpful as algae
can concurrently remove nutrients and TMs (Almomani et al., 2019b;
AlMomani and Örmeci, 2016; Judd et al., 2017; Znad et al., 2018).
4. Conclusion
The present study proposes and optimizes a process for the removal
of toxic metals (Ni, Al and Cu) from industrial wastewater metals using
Spirulina platensis and Chlorella vulgaris algae strains. The removal effi-
ciency of algae strains was improved by applying acidic treatment
(AC-T) using sulfuric acid. The AC-T improved the surface properties
of the algae strains, increased the portion of the negatively charged
functional groups on the surface of algae, and achieved a significant %
TMrem. The removal efficiencies of Al, Ni and Cu attained with acid
treated SP were ≥95.0%, 87.0%, and with treated CV were 63.0%, and
87.0, 79.1, and 80.0%, which are 1.8 and 1.5 folds higher than untreated
strains. The bio-sorption of TMs on SP and CV is spontaneous, exhibits a
monolayer mechanism, and follows second-order kinetics. Combined
electrostatic attraction forces and ion-exchange processes control and
establish the main mechanism between the algae strains and TMs. The
SRM algorithms revealed that contact time, pH, and bio-sorbent dose
are the main performance parameters for maximum removal efficiency
under optimum operational conditions.

60 Funding

50
Open Access funding provided by the Qatar National Library.
40
5.5 CRediT authorship contribution statement
5.0 8.0
7.0
4.0 6.0 Fares Almomani: Conceptualization, Methodology, Modeling,
C 3.0 5.0
alg Figures preparation, software, pre-Writing- Original draft preparation,
ae (g/ 2.0 4.0
L) 1.0 3.0 PH Data curation, Discussion, Validation, Tables preparation, Writing-
0.5 2.0 Reviewing and Editing.
Rahul R. Bhosale: pre-Writing- Original draft preparation, Data
curation, Discussion.

Declaration of competing interest

100
(b) There are no known conflicts of interest associated with this
90 publication.
80
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.0
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