Effect of Pretreatment of Biosorbent in Biosorption: A Comparative Study

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 3, Pages: 894-899

J. Environ. Treat. Tech.

ISSN: 2309-1185

Journal web link: http://www.jett.dormaj.com

Effect of Pretreatment of Biosorbent in Biosorption:

A Comparative Study

Ariani Dwi Astuti ¹^,², Khalida Muda ¹^*

Department of Water and Environmental Engineering, School of Civil Engineering, Faculty of Engineering, Universiti Teknologi Malaysia, Johor,

Malaysia

Department of Environmental Engineering, Faculty of Landscape Architecture and Environmental Technology, Universitas Trisakti, Jakarta, Indonesia

Received: 21/01/2020

Accepted: 24/05/2020

Published: 20/09/2020

Abstract

The textile industry generates large amounts of wastewater with strong BOD/COD and salt load, which are often seen in dark colour. An

alternative treatment for this type of wastewater is the biosorption, where it involves a passive uptake of both the organic and inorganic

compounds, including dye or its derivatives, using non-growing/living microbial mass. Biosorbent pretreatment, autoclaving techniques and

combined with the chemical processes, such as acidification, were discussed. The response surface methodology (RSM) is used for

researching and developing the effect of pH, contact time, dosage, and biosorbent size in the biosorption process in synthetic textile

wastewater using Bjerkandera adusta. When the pH was 4, and the contact time, biosorbent dosage and biosorbent size were 90 minutes,

000 mg/L and 0.4 mm, respectively, the optimal removal circumstance was able to be verified, at 53.55%, and 81.3% of colour removals

were demonstrated through the experimental procedure. This leads to the high acceptance of the experimental findings and model forecast.

adj

In the optimisation of experimental parameters, the quadratic model estimated both R and R correlation coefficients quite satisfactorily

as 0.988, 0.977, 0.926, and 0.783, respectively. It is more effective to combine the autoclaving technique with chemical processes than

adopting just the autoclaving method. The two-sided t-test was used to identify any significant variations in the preparation techniques of

biosorbents using p < 0.05. The biosorbent study using scanning electron microscopy (SEM) and characterisation of surface functional group

using Fourier-transform infrared (FTIR) spectroscopy confirms the results obtained.

Keywords: Bjerkandera adusta, Synthetic textile wastewater, Biosorbent, Response surface methodology, Colour removal efficiency

Introduction¹

sectors (8). Various techniques for treating textile waste are being

adopted, including physical-chemical methods, such as

adsorption, filtering, biological processes, chemical flocculation,

coagulation, and sedimentation (9). Although most of these

approaches are highly effective, some are rather expensive and

produce sludge that requires further treatment (e.g., sludge

treatment). The ability of fungi to remove a large selection of

contaminants that are intransigent, such as synthetic dyes, has

been approved (10,11). The highest rate of degradation was

recorded for Bjerkandera adusta, whose strain could completely

decolourise a large number of colourants and detoxify synthetic

wastewaters (7). Bjerkandera adusta, Phanerochaete

chrysosporium, and Pleurotus ostreatus exhibited the best

decolourisation properties (12), while Heinling et al. (1997) stated

that Phanerochaete chrysosporium, Trametes versicolor, and

Bjerkandera adusta had demonstrated their capability to decolour

all of the colours tried (13). Through its production of ligninolytic

enzymes, Bjerkandera adusta indicated that this strain could be

beneficial for biotechnological advancement (14). Some methods

of pretreatment may enhance the adsorption capacity of biomass

materials. Other techniques of pretreatment include autoclaving,

The textile dyeing process uses a large amount of water,

making it one of the biggest liquid pollutant generators (1, 2).

Towards the turn of the decade, the world population is estimated

to be 11 billion due to its exponential growth from only 1 billion

in the 1800s to more than 7 billion today. With a per capita intake

of textiles and clothing of 7 kg per person per year, the global

annual demand for textile products has exceeded 49 billion kg (3),

where the discharge is between 115–175 kg COD/ton of

completed textile (4). Manufacturing license in Malaysia

(Malaysia Investment Performance, MIDA, 2011) has

documented a total of 662 licensed textile and 1,000 clothing

mills on a small scale. The treatment of wastewater produced by

textile companies creates an important environmental challenge,

particularly in terms of the expense of quality of the resulting

textile effluent (5). During the dyeing process, textile dyeing

factories use adequate quantities of different colours or additional

chemicals and ultimately emit robust textile wastewater (6,7).

Azo dyes currently reflect the better standard of synthetic dyes by

global standards (around 70 percent), and their large-scale

production may be related to their use by various manufacturing

Corresponding author: Khalida Muda, Department of Water and Environmental Engineering, School of Civil Engineering, Faculty of

Engineering, Universiti Teknologi Malaysia, Johor, Malaysia. E-mail: Khalida Muda. Email: khalida@utm.my.

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 3, Pages: 894-899

which might break the structure of fungi and reveal the potential

colour of the binding sites (15).

(21). After stirring the flask for predetermined time intervals, the

flasks were extracted and resuspended to obtain the liquid-solid

separation prior to its analysis. In addition to the biosorbent used

in this work, the SEM (TM3000 Hitachi High Technologies

America, USA) was used to study the morphological structure,

whereas FTIR spectroscopy (Nicolet iS10 FT-IR Spectrometer,

USA) was used to investigate the chemical characterisation.

Response surface methodology (RSM) was chosen as the

analytical instrument for optimising multiple variables and

predicting better performance. The method allows the

determination of optimal conditions under predetermined factor

preferences, such as high removal at the optimal condition and/or

with the least number of experiments (16).

The main aim of this research is to assess the effects of pH,

biosorbent dosage, contact time, and biosorbent size in the

biosorption process in synthetic textile wastewater using

Bjerkandera adusta via two pretreatment methods of biosorbent;

for instance, the process of autoclaving and combining

autoclaving with chemical processes. RSM was applied, as it was

among the most sophisticated and appropriate variable techniques

used in analytical advancement. RSM was used for planning

experiments, building models, and defining the optimal

conditions.

2.4 Experimental Design

RSM was used to determine the relationship between colour

removal and the critical variables (e.g., pH, biosorbent dosage,

contact time, and biosorbent size). In this experiment, a four-

factor with a five-level CCD was used, which required almost 30

studies. The factors were: A: pH (4–8); B: biosorbent dosage; C:

contact time (30–150 minutes); and D: biosorbent size (0.2–0.6 ±

0.05 mm). The experiments were selected randomly for statistical

evaluations, and every sample was duplicated using a software

package called the Design-Expert Version 11 (Stat-Ease,

Statistical Made Easy, MN, USA).

Methods

.1 Synthetic Textile Effluent

3 Results and Discussion

The following contents of the synthetic wastewaterwere used

The RSM can be in the form of a variety of mathematical

and applied mathematical approaches. RSM promotes the

practice of experimental data-based polynomial equations,

which must characterise the knowledge conduct set with the

goal of creating relevant mathematical forecasts (22). The

expected value of information was achieved in this research

by using the Design-Expert software to fit the model

technique. After combining this information with distinct

models such as cubic, ANOVA demonstrates that all responses

were better represented by the mathematical quadratic

polynomial model (Table 1).

to test the performance: K

HPO

0.58 g/L, NH Cl 0.16 g/L,

KH PO 0.23 g/L, MgSO ⋅7H

O 0.09 g/L, CaCl ⋅2H 0 0.07 g/L,

trace solution 1 ml/L, and EDTA 0.02 g/L. The carbon sources

considered in this experiment were: sodium acetate (0.5 g/L),

glucose (0.5 g/L) and ethanol (0.125 g/L). The trace element was

composed of FeCl ⋅4H O (1.5 g/L), H BO (0.15 g/L),

MnCl ⋅4H O (0.12 g/L), ZnCl (0.12 g/L), CoCl ⋅6H O (0.15

g/L), CuCl ⋅2H O (0.03 g/L), NaMoO ⋅2H O (0.06 g/L), and KI

.03 g/L (17). For this study, the azo dyes were mixed at a

concentration of 60 mg/L consist of Reactive Black 5, Reactive

Blue 4 and Disperse Orange 2.

The predicted R value for pretreatment method A of 0.9345

is based on the rational consensus with the adjusted R value

equivalent to 0.9776; this implies that the difference is below 0.2

.2 Pretreatment of Biosorbents

The Bjerkandera adusta fungi were cultivated in a static

(23). In addition, sufficient accuracy also tests the signal-to-noise

ratio. A ratio of above 4 is ideal in this circumstance, where it sets

a satisfactory signal by a ratio value equivalent to 37.058. This

model can, therefore, be used to manage research methods (Figure

condition on malt extract agar (prepared in the laboratory

following the DMZS media number 90 description) at a pH of 5.6

and a temperature of 25°C. At the end of 7 days (18), when

sporulation had taken place, the fungal biomass was autoclaved

to eliminate the fungal biomass at 121°C and 103.42 kPa for 45

min (19). Pretreatment method A (autoclaved) the autoclaved

non-viable biomass was properly washed with distilled water and

dried for 36 hours at 70 ± 1°C in the oven. Then, the dried fungal

biomass was crushed into a powder form and sieved to a granular

size between 0.2–0.6 mm. Next, it was transferred to 50 mL of

). The predicted R value of 0.7820 is also reasonably under the

arrangement, with the adjusted R of 0.9266 for pretreatment

method B, at which the difference is less than 0.2. The 21,461

ratio shows an appropriate signal; hence, this model is suitable to

be used for navigating the design room.

The highest colour removal was 53.55% and 81.3% for

biosorbent preparation methods A and B, respectively, based on

the experimental results. This condition is achieved when the pH,

contact time, biosorbent dosage, and biosorbent size were 4, 90

minutes, 3000 mg/L, and 0.4 mm, respectively. The primary

means of comparison is the percentage difference between

biosorbent preparation methods A and B. A research by Farah., J

et al. (2007) using baker’s yeast as iosorbent (dried-autoclaved

pretreatment) has achieved approximately 80% removal of

Astrazon Blue in a contact time of 4 h, shaking speed of 150 rpm,

biosorbent dosage of 6600 mg/L, initial pH of 7, and biosorbent

size of 0.210 mm (24).

.1 M HCl of autoclaved non-viable biomass (about 10.0 g) for

pretreatment method B (a combination of autoclaved-chemical

methods), and the mixture was stirred at 250 rpm and room

temperature for 1 h (20). Subsequently, the non-viable acid-

treated biomass was appropriately washed with distilled water,

and then powdered in pretreatment method A.

.3 Batch Testing

A batch testing was conducted by stirring 50 ml of synthetic

textile wastewater together with the biosorbents in a flask at a

room temperature of 25°C ± 2°C and shaking speed of 160 rpm

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 3, Pages: 894-899

Table 1: Variance analysis (ANOVA) and model statistics discovered using Bjerkandera Adusta from Dye Biosorption Process

Pre-treatment Methods A*)

Pre-treatment Methods B

Source

Sum of

square

Mean

Sum of

square

Mean

F-value

P-value

F-value

P-value

square

301.69

2953.9

square

520.18

4746.9

Model

A-pH

4223.68

2953.93

91.5

< 0.0001

7282.5

4746.9

27.14

< 0.0001

895.88

247.70

B-Weight

Biosorbent

C-Contact

Time

D-Size of

Biosorbent

70.57

570.57

14.41

84.15

173.04

4.37

< 0.0001

0.054

444.71

37.68

444.71

37.68

23.20

1.97

0.0002

0.1812

0.0034

4.41

4.15

25.52

0.0001

231.57

12.08

103.43

1.69

103.43

1.69

31.37

0.5125

20.02

0.2089

2.3

< 0.0001

0.485

202.71

14.04

154.44

12.66

19.87

7.22

202.71

14.04

154.44

12.66

19.87

7.22

10.58

0.7328

8.06

0.0054

0.4054

0.0124

0.4291

0.3247

0.5485

< 0.0001

0.4954

0.4478

0.5995

66.02

0.6889

7.59

66.02

0.6889

7.59

0.0004

0.6542

0.15

0.6604

1.04

0.555

412.39

1.71

0.555

412.39

1.71

0.1683

125.07

0.5199

0.0853

3.34

0.6874

< 0.0001

0.482

0.3769

66.95

0.4882

0.6076

0.2878

A²

1283.1

9.36

1283.1

9.36

B²

C²

0.2812

11.02

49.46

48.35

1.11

0.2812

11.02

3.3

0.7743

0.0875

11.64

5.52

11.64

5.52

D²

Residual

Lack of Fit

Pure Error

Cor Total

287.47

286.24

1.23

19.16

28.62

0.2454

4.83

21.76

0.0017

116.63

< 0.0001

0.2221

4273.14

7570.0

*) Astuti and Muda, (2018)

(a)

(b)

Figure 1: Graph of The Predicted Vs. Actual Color Removal Process Using Bjerkandera Adusta (a.) Methods A and (b.) Methods B

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 3, Pages: 894-899

(a)

(b)

Figure 2: (a) Methods A and (b) Methods B biosorbent process magnification powers (8000X)

Figure 3: FTIR spectrum of biosorbent Bjerkandera adusta Methods A and Methods B

The initial concentration of Astrazon Blue (single pure dye)

Worku and Sahu (2014) had also carried out the colour

removal work with various adsorbent pretreatment methods using

rice husk. It was found that the maximum dye removal was 94%

at pH 2; adsorbent dose 50,000 mg/L; dye initial concentration

used was 100 mg/L. Meanwhile, Khalaf ‘s research findings

showed that the dried-autoclaved pretreatment of biomass A.

niger and Spirogyra sp. has a total dye removal of 88% and 85%,

respectively, at a pH of 3, the temperature of 30°C and biosorbent

dosage of 8000 mg/L after an 18-h contact time (25). Compared

with the findings of this study, other researchers required higher

doses of biosorbents and longer contact time.

100 mg/L; and

a contact time 160 minutes for the

physicochemical treatment of rice husk and 73% at an adsorbent

dosage of 50,000 mg/L, pH 2, dye initial concentration 50 mg/L,

and 80 minute of contact time for the physically treated rice husk

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 3, Pages: 894-899

(26). The colour removal is quite high compared to this result, but

indicates aliphatic groups (symmetric and asymmetric

it required a higher dose of adsorbent (i.e., 50,000 mg/L).

However, a research conducted by Worku and Sahu is in line with

the findings of this study, where the adsorbent pretreatment

method had resulted in higher colour removal. This result is

consistent with the results of previous studies, whereby it was

learnt that autoclaving can break down the structure of fungi and

expose the potential binding sites for dyes, while the pretreatment

of acid can change the surface of negatively charged fungal

biomass to become positively charged and thus, increasing the

attractiveness between the fungal biomass and the

colourants/dyes (15).

A t-test is a type of descriptive statistics used to assess

whether the two group means differ, which can be attributed to

specific characteristics. Two-sided t-tests were used to identify

any crucial differences between the preparation methods,

autoclaving, and autoclaving techniques in combination with the

chemical process, and colour removal efficiencies at p < 0.05. The

findings showed a significant difference between the means of the

methods, as the value of t calculated had exceeded the t table.

The SEM analysis was carried out to clarify the

morphological characteristics, along with the surface

characteristics of the adsorption samples. The analysis also

modelled the surface porosity and structure/texture of the

processed biosorbent. In addition, the adsorption capacity relies

on the particle size, distribution of pores, and surface area or

porosity of micro/mesoporous materials (27).

stretch/CH

vibration). A cluster of water molecules was assigned

to free the hydroxyl on biosorbents Bjerkandera adusta

-1

represented by the peak at 1640 cm . The peak at 1160 cm may

imply the presence of a C-O stretching band of alcohol, hydroxyl,

or ether. The intensity of the peaks gradually increases in the

pretreatment of combining autoclave and chemical processes.

-1

Some new peaks were detected at 1800 cm and 1450 cm ;

this datum shows that in pretreatment method B, the carboxyl

groups have appeared, implying that the combination process has

succeeded. The findings indicate that the biosorption process was

affected by the presence of functional groups on the biosorbent

Bjerkandera adusta surface. The results of this research are

similar to those outlined by Mahmoud (28) and K. Jain (30). The

FTIR and SEM analyses also confirmed the dye-biosorbent

interactions and found that functional groups such as carboxyl,

amine, amide, and hydroxyl on the surface of biosorbent were

liable for the biosorption of a reactive dye (RB49) (31).

Conclusion

A study was carried out to investigate the effects of pH,

biosorbent dosage, contact time, and biosorbent size in the

biosorption process with Bjerkandera adusta in synthetic textile

wastewater using two methods of biosorbent pretreatment, i.e.,

autoclaving and combination of autoclaving with chemical

processes. Research findings showed that variables, such as pH,

biosorbent dosage, and biosorbent size, were the key factors that

influenced colour removal in the biosorption with Bjerkandera

Adusta. Experimental values are as expected under optimal

conditions, demonstrating the suitability of the model and the

achievement of RSM in optimising the state of the biosorption

process using Bjerkandera adusta. The two-sided t-test was used

with p < 0.05 to identify the important differences in the

biosorbent pretreatment technique, in which it was found that the

best biosorbent preparation method was the combination of

autoclaving with chemical processes. The assessment of SEM and

FTIR analyses has led to the conclusion that the colour removal

technique was conducted via the biosorption process.

Biosorbent Bjerkandera adusta had undergone the SEM

analysis at a power magnification of 8000× and 2000× before and

after the biosorption process, respectively. After some interaction

with the pollutant dye, the controlled and treated biosorbents were

monitored and confirmed to have experienced some changes,

such as surface structure and size of pores. The size of the

biosorbent pores Bjerkandera adusta method A was between

.74–8.30 μm and changed to 1.31–2.14 μm after biosorption.

Meanwhile, the size of the biosorbent pores Bjerkandera adusta

method B had varied from 2.55–4.81 μm and became 1.14–4.25

μm after biosorption. The dye deposits between biosorbents

Bjerkandera adusta are indicated as crystalline. Therefore, the

SEM results showed the most excellent bond with the colour

being explored. These findings support the previous research by

Mahmoud et al. (2017) using baker’s yeast (28). The biosorbent

pores B. adusta method B has a pore size smaller than those in

method A. Small pore size expands the surface of the adsorption

to produce better colour removal (Figure 2).

Acknowledgment

The authors wish to thank Universiti Teknologi Malaysia for

the financial support given to this research (Grant No. 18H96).

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