Construction and Building Materials 167 (2018) 813–821

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Construction and Building Materials

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A novel study on using protein based biopolymers in soil strengthening

Hadi Fatehi a, Sayyed Mahdi Abtahi a, Hamid Hashemolhosseini a, Sayyed Mahdi Hejazi b,⇑
a
Department of Civil Engineering, Isfahan University of Technology, Isfahan 84156-83111, Iran
b
Department of Textile Engineering, Isfahan University of Technology, Isfahan 84156-83111, Iran

h i g h l i g h t s

Casein and sodium caseinate biopolymers were used to stabilize dune sand.

UCS, CBR, direct shear and leaching tests were conducted on stabilized samples.

Micro structure was studied by SEM method and a descriptive model was developed.

The modifying effect of casein was compared with gellan, agar, xanthan and lignin.

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

Article history: New protein-based biopolymers are introduced in this study to stabilize dune sand. Since the conven-
Received 18 June 2017 tional soil stabilization materials, especially cement, have harmful effects on the environment, alternative
Received in revised form 21 October 2017 eco-friendly materials, casein and sodium caseinate salt biopolymers have been used in this study to
Accepted 6 February 2018
reduce environmental concerns. Casein and sodium caseinate biopolymers obtained from milk were
added to dune sand, and mechanical properties of biopolymer treated sand were investigated through
a series of laboratory tests. The effects of biopolymer content, curing time, curing temperature, and fat
Keywords:
milk content on improving sand were studied by unconfined compression test. The results showed that
Protein based biopolymers
Soil strengthening
compressive strength of biopolymer treated sand was increased when the curing time had passed, and
Compressive strength biopolymer content increased. Moreover, temperature can be effective on improving compressive
Biopolymer–soil interaction strength of samples treated by casein and sodium caseinate. The positive influence of curing temperature
Sandy soil on compressive strength of modified soil was estimated to be up to 60 °C. In addition, direct shear test,
leaching test and California bearing ratio (CBR) test alongside microscopic observation by scanning elec-
trons microscopy images (SEM) were performed. On the whole, the protein-based biopolymers propose a
strong potential as additives for soil treatment rather than some material such as cement and chemical
polymers.
Ó 2018 Elsevier Ltd. All rights reserved.

1. Introduction cement, it has caused numerous detrimental impacts on the envi-

Soil treatment is an essential concern for geotechnical engineers
because of urbanization and population growth throughout the
world. Generally, ground improvement aims to resolve various
geotechnical problems such as reducing differential settlements
of foundations, improving mechanical parameters of subgrade lay-
ers, consolidation and compaction of the soil, and the improvement
of shear strength [1]. For these purposes, various methods and
materials have been used till now. Among them, cement is the
most widely used additive owing to adequate strength, availability,
and low cost [2]. In spite of advantages and various applications of
⇑ Corresponding author.
E-mail address: hejazi110@cc.iut.ac.ir (S.M. Hejazi).
ronment [3].
It should be noted that during production process of cement,
about 0.95 ton CO2 per ton of cement is imposed into the atmo-
sphere [4]. Whereas, the rate of CO2 production by cement indus-
tries has risen in the last few years from 4.2% in 1980 to 9.0% in
2012. The proportion of geotechnical projects is also almost 2% of
total CO2 emissions produced by cement [3]. Global warming is
one the harmful effects of increasing the amount of greenhouse
gases [5]; besides, glacier melting and rising sea level [6], intense
storms and tornados, and drought are all consequences of global
warming [7]. Moreover, changing the soil pH, urban runoff, pre-
venting the surface vegetation growth, and concrete dust are other
environmental drawbacks of using cement [3].
Therefore, in the current century, environmental concerns have
led to the use of different eco-friendly methods and materials in

https://doi.org/10.1016/j.conbuildmat.2018.02.028
0950-0618/Ó 2018 Elsevier Ltd. All rights reserved.
814 H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821
geotechnical activities. For this, useful bacteria and enzymes have
been employed in endeavors towards soil improvement [3].
Although the microbial method has been satisfying with accept-
able performance, some factors make this procedure complicated.
In this manner, numerous environmental parameters such as tem-
perature, pH, concentration of electron donor and electron accep-
tor, and the rate of nutrients and metabolites are simultaneously
effective to gain adequate achievement. Moreover, microbial soil
treatment is slightly more time-consuming compared with other
methods [8].
A biopolymer is a polymeric substance (as a protein or polysac-
charide) formed in a biological system [9]. The use of biopolymers
in civil engineering activities dates back several years ago. But, it
has been considered more frequently in recent years by civil engi-
neers [3]. Lignin is the most abundant organic polymer existed on
earth after cellulose [10]. The influences of Lignin, which is the
third largest part of agricultural biomass, have been studied in dif-
ferent geotechnical aspects such as constitutive model for
lignosulfonate-treated soil [11], thermal, mechanical, and
microstructural characteristics of lignin treated soil [12–15], and
Transportation infrastructure [16–18]. The optimum moisture con-
tent of lignin for improving silty soil was reported 12% by Zhang
et al. Gaining the maximum compressive strength of lignin treated
silt required 28 days of curing, also lignin changed the physico-
chemical behavior of sand minerals to provide a resistant lignin
treated silt [12].
In recent years, new ecological materials have been introduced
to reduce hazard environmental impacts of traditional additives; in
this regard, two thermo-gelation biopolymers (gellan gum and gar
gum) were evaluated by Chang et al. (2012)., which enable hydro-
gen bonds to enhance the strength and durability of sandy and
clayey soils. While time was not a crucial parameter, thermal con-
ditions played a very important role in soil treatment [19]. Chang
et al. also introduced xanthan gum and b-1,3/1,6-Glucan biopoly-
mer to improve the geotechnical behavior of different types of soils
[20,21]. Furthermore, many other biopolymers including tannin
[22], Alginate [23], Guar Gum [24], and starch indicate that the
use of biopolymers would be operational and effective in soil
strengthening. The studies show that biopolymers could be consid-
ered an environment-friendly alternative for soil improvement and
mixed directly with soil, so this may resolve the need for providing
complex conditions regarding microbial injection method [8].
In this study, Casein and sodium caseinate salt biopolymers are
introduced as new soil additives to improve the geotechnical
behavior of poor graded sand. How to produce Casein biopolymer
from milk and its chemical characteristics are also described. Fur-
thermore, the mechanical and geotechnical behavior of sand trea-
ted by Casein is investigated through a series of laboratory
experiments. Besides, the interaction between soil and casein par-
ticles is represented by SEM images.
2. Materials and methods
2.1. Casein and sodium caseinate biopolymers
Casein is a protein-based biopolymer making up 80% of the pro-
teins of cow’s milk [25]. Casein has been derived from the Latin
word ‘‘Caseus” which means cheese [26]. The isoelectric point of
casein is about 4.6 and generally, the casein pH is 6.6 and has a
negative charge. Casein chemical formula is C31 H27 NO4 [27].
Casein and sodium caseinate salt as the additives have been used
in this paper. When milk becomes sour, it separates into curd
and whey are prepared, now by separating the water, casein is
introduced. Casein can also be extracted from the milk by reducing
the milk pH and adding a mineral acid into the milk [28].
The best quality of casein is gained when the fat milk and acid
amount are in the lowest content [29]. So that the skim milk was
used for this purpose. Then, the casein were washed after separat-
ing it from the milk to reduce the acid content. Casein, with its
cheesy state, is insoluble in water. When sodium hydroxide is
added to casein, the reaction between them produces sodium case-
inate salt, which is a water soluble glue [27]. The produced mois-
ture causes the sodium caseinate to be in the form of a paste and
more appropriate for making a homogenous mixture compared
with the cheesy texture of casein (see Fig. 1).
2.2. Soil
Iran is a country with an arid climate and covered by deserts
such as Dasht-e Loot and Dasht-e Kavir. So that facing to dune
sands of deserts, which categorized as poor soils, environment in
constructional projects is inevitable. Uniform soil gradation, lack
of cohesion among soil particles, high settlement, and low geotech-
nical strength are the main problems of dune sand. So, owing to the
immense amount of dune sand throughout the world, improving
the geotechnical parameters of loose granular soils and dune sand
is essential.
In this study, the dune sand was collected from a highway con-
struction site in Kerman, Iran. According to Table 1, the optimum
moisture and maximum dry density contents were 1.74 g/cm3
and 16%, respectively obtained from the standard Proctor com-
paction method based on ASTM D2216 [30]. Sieve Analysis were
performed and the results are presented in Fig. 2. Since more than
95% of sand grains were larger than 0.075 mm and smaller than 2
mm, the soil was a poorly graded sand (SP) according to the Unified
Soil Classification System (USCS). As material is transported, it is
subject to abrasion and impact with other particles which tends
to ‘‘round-off” the sharp edges or corners. This can be seen in
Fig. 9a. the soil structural composition is between emin = 0.6 and
emax = 0.78, the specific gravity (Gs) was also 2.63. Based on XRD
test results (Table 2), the soil includes different types of materials:
silica (SiO2) by 55.66%, thallium sulfide (Tl2S) by 13.13%, InO3 by
11.39%, Ba3Ta2ZnO9 by 4.97%, Rb6Te2O9 by 9.63%, and Li9KNb10O30
by 5.22%.
2.3. Specimen preparation
The soil was mixed with different weight ratio of casein and
sodium caseinate. For all experiments, three tests were repeated
for each sample to ensure that the results are accurate and the
average values were reported for each treatment, and the variation
with average value was considered less than 5%. The biopolymer-
sand mixture placed into the special molds; for unconfined com-
pression test, the molds were 36 mm in diameter and 75 mm in
height based on ASTM D2166 and were made of polypropylene
to prevent samples sticking to the walls. Furthermore, a linear
groove was cut into the mold to extract samples more easily. While
the specimens were being made, this groove was blocked by two
pipe fasteners at the top and bottom of molds. Upper and lower
surfaces were slightly trimmed to avoid non-uniform distribution
of stress. In addition, two filter papers were put on the top and bot-
tom surfaces to prevent stress localization. Direct shear test spec-
imens also were fabricated in the metal molds and compacted at
two layers that were 20 mm in height and 63 mm in diameter.
All specimens were prepared by optimum moisture content and
maximum soil dry density.
2.4. Experimental program
The dune sand was completely air dried so that the designed
amount of additives and water were measured based on dry den-
 H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821 815

Fig. 1. a) Casein biopolymer, b) Sodium caseinate salt biopolymer.

Table 1
Basic properties of the dune sand used in this study.

Soil type D50 (mm) Cu Cc Gs Shape USCS emax emin OMC (%) Maximum Dry Density (g/cm3)
Sand 0.18 2.2 0.77 2.63 Round SP 0.78 0.6 16 1.74

specimens. The amount of fat in milk also affects on adhesion

100 between biopolymer and soil particles. Therefore, casein and
90
sodium caseinate obtained from skim milk and full fat milk (3%
Percent Finer by Weight (%)

80
fat) were used to investigate the role of fat cells on the compressive
70
strength of biopolymer treated sand.
60
50
40
2.4.2. Direct shear test
30 The direct shear tests were carried out based on ASTM D3080
20 [32] to determine the consolidated drained shear strength of
10 biopolymer treated soil. Although direct shear test is one of the
0 easiest methods to obtain the shear strength, the predetermined
0.01 0.1 1 10 fracture plane is considered as a technical problem [33]. Since dune
Grain Size (mm) sand particles were rounded and uniform in size, the influence of
this problem could to some extent be ignored. The direct shear
Fig. 2. Particle size distribution curve of the sand used in this study.
strength tests were carried out by applying a constant strain rate
of 1.06 mm/min on the soil samples put inside standard shear boxes
sity of soil and optimum moisture content. The unconfined com- until the soil had failed or reached a maximum horizontal displace-
pression test, direct shear test, and california bearing ratio (CBR) ment of 10 mm. In addition, three different normal stresses were
test were performed on both neat and modified soil samples. applied on specimens including 57.6, 90.77, and 123.9 kPa.

2.4.1. Unconfined compression test 2.4.3. California bearing ratio test

The uniaxial compressive tests were conducted in accordance
with ASTM D2166 [31] using standard UTM (Universal Testing
Machine) device. The influences of additive amount, temperature,
time, and milk fat content on the biopolymer-treated soil samples
were investigated by uniaxial compressive tests. For biopolymer
content parameter, the various amounts (0.5, 1, 2, 3, and 5%) of
casein and sodium caseinate were added into the soil. Moreover,
the compressive strength of biopolymer (1%) treated sand was
measured at 25, 40, 60, 80, and 120 °C to determine the best curing
temperature. The variation of uniaxial compressive strength during
the time was studied in 3, 7, 14, and 28 days after fabricating the
The california bearing ratio (CBR) tests were performed accord-
ing to ASTM D1883-16 [34] to study CBR strength of casein and
caseinate treated soil for pavement subgrade and subbase applica-
tions. Fig. 2 shows that all soil particle sizes were less than 19 mm
(3/4 in). The specimens were made and compacted at optimum
moisture content, the biopolymer additive and water content were
added to the soil based on the maximum dry density. Three repe-
titions were carried out to reduce experimental mistakes. The pro-
portion of biopolymers used was 0.5% and 1% which were added to
the sand and cured for 14 days and the unsoaked CBR tests were
conducted.

Table 2
Oxide chemistry of dune sand.

Oxide Chemistry SiO2 Tl2S InO3 Ba3Ta2ZnO9 Rb6Te2O9 Li9KNb10O30

Characteristic (%) 55.66 13.13 11.39 4.97 9.63 5.22
816 H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821

2.4.4. Leaching test
In order to understanding the strength of the stabilized soil
against water flow, the leaching test was performed. For this pur-
pose, pure soil and sand treated by 2% of biopolymers were used.
These samples were made in the molds with 120 mm height, 50
mm external diameter and 45 mm internal diameter. A porous
sheet put at the end of the molds to prevent the soil particles from
escaping. Blocking the end of samples against water passing, 2
days waited for each sample to become completely saturated.
Then, water from the height of 3 m was passed through the speci-
mens. Finally, the amount of protein dissolved in water passed
through the samples was measured by Kjeldahl method (see
Fig. 3).
2.4.5. Microscopic observation
Microstructural analysis of biopolymers treated sand samples
were investigated through scanning electron microscopy (SEM)
images. It is worthy to note that the images were taken from the
center of fractured UCS samples by Philips XL30, 1994 scanning
electron microscope. This analysis provides information about
how casein and sodium caseinate biopolymer interact with soil
particles. Dune sand samples containing 1% and 2% biopolymers
were selected and after ensuring that the samples were completely
dried, the surfaces were coated with a conductive metal (gold) to
avoid electron scattering. For the sake of completeness, the
biopolymer content, curing time, and curing temperature of the
treated soil, subjected to various tests, are summarized in Table 3.
3. Results and discussion
3.1. Unconfined compressive strength results
The uniaxial compressive strength of casein and sodium casei-
nate treated sand were investigated in terms of biopolymer con-
Fig. 3. Samples prepared for leaching test.
tent, curing temperature, and curing time. Fig. 4 shows how
unconfined compressive strength changes at different contents of
biopolymers. Both casein and sodium caseinate multiplied the
compressive strength of soil samples which also have a tendency
to increase with an increase in the biopolymer content. The
untreated dune sand demonstrated low compressive resistance
(21 kPa) owing to uniform grading and lack of angular and fine-
graded particles. 0.5% casein and sodium caseinate improved the
compressive strength up to 450 kPa. For higher proportions, the
mixture of sand with sodium caseinate represented a better perfor-
mance because the pasty shape of sodium caseinate in comparison
to the cheesy state of casein biopolymer provides better conditions
to produce a homogeneous mixture. The compressive strength of
sodium caseinate treated soil samples were 17% and 9% higher
than the samples stabilized by casein for 3% and 5% additives,
respectively. An example of fractured unconfined compression
specimens are shown in Fig. 5.
The effect of curing time on the compressive strength of casein
and sodium caseinate (1%) treated sand was investigated in 3, 7,
14, and 28 days of curing. Fig. 6 shows the comparable strength
values obtained for casein and sodium caseinate treated soil during
the time. The compressive strength of casein treated specimens has
been significantly grown until 7 days. The strength reached almost
99% of its maximum which was obtained after 14 days of curing,
while the increasing process remained constant up to 28 days.
On the other hand, the high moisture content of sodium caseinate
treated sand samples prevented us from performing unconfined
compression test until the third day of curing. After the third
day, the compressive strength experienced a considerable increase
until the seventh day and kept increasing to its maximum value at
14th day. It can be also observed that more time is needed for
sodium caseinate treated soil to obtain maximum compressive
strength compared with casein treated soil because when the
sodium hydroxide is added to casein, sodium is replaced with
hydrogen and consequently sodium caseinate and water would
be produced so that the moisture content is more than the casein
and the curing time is expected to take longer.
The curing temperature is considered as an important parame-
ter influencing on stabilization of soil by different biopolymers.
Therefore, the compressive strength of casein and sodium casei-
nate treated sand cured at various temperatures (25, 40, 60, 80,
and 120 °C) has been investigated. All specimens were made using
1% biopolymer and were cured for 14 days. Fig. 7 shows strength-
ening behavior of casein and sodium caseinate treated sand cured
under different thermal conditions. It can be seen that compressive
strength improved when the temperature rose up to 60 °C for both
biopolymers. The higher temperatures had a significant effect to
weaken compressive strength and adhesive forces between
biopolymer and soil particles. For curing temperature of 120 °C

1800
Table 3 1600
Experiments program. Note: UCT, unconfined compression test; DST, direct shear 1400
test; CBR, california bearing ratio; SEM, scanning electron microscopy.
1200
UCS (kPa)

Test type Biopolymer Curing Time Curing Temperature

1000
content (%) (days) (C)
800
UCT 0, 0.5, 1, 2, 3, 5 14 21–25
1 3, 7, 14, 28 21–25 600
1 14 25, 40, 60, 80, 120 400
1a 14 21–25
200
DST 0, 0.5, 1 14 21–25
CBR 0, 0.5, 1 14 21–25 0
0 1 2 3 4 5 6
Leaching Test 0, 2 14 21–25
SEM 1, 2 14 21–25 Biopolymer content (%)
a
1% casein and sodium caseinate made by full fat (3% fat) milk and skim milk was Fig. 4. The compressive strength variation with different casein and sodium
used to investigate the influences of fat cells. caseinate contents.
 H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821 817

Fig. 5. Fracture of unconfined compression test samples a) casein treated sand b) sodium caseinate treated sand.

caused casein and sodium caseinate to interact more effectively

700
and to make more resistant bonds with soil particles, whereas cur-
600 ing at higher temperatures (80 and 120 °C) gradually caused casein
and sodium caseinate particles to be decomposed to amino acids
500
which leads to reduction in the compressive strength. Therefore,
UCS (kPa)

400 it can be concluded that thermal treatment is an essential and

viable factor for soil improvement using casein and sodium casei-
300
nate biopolymers.
200 Fat cells can also influence on strength of casein and sodium
caseinate treated soil. The highest quality of casein biopolymer is
100
obtained when fat cells are emulsified through the milk completely
0 [29]. The casein and sodium caseinate produced from skim milk
0 5 10 15 20 25 30 and full-fat milk (3%) were used to study the influence of fat milk
Time (day) amount on compressive strength of biopolymer treated sand.
Table 4 indicates that fat cells besides casein compound consider-
Fig. 6. comparison of strengthening behaviors of casein and sodium caseinate
ably reduced the strength, whereas there was a 28% decline from
treated soil during the time.
596.76 kPa to 427.64 kPa for casein treated sand and 22% decline
from 635.34 kPa to 495.18 kPa for sodium caseinate treated sand.
Herein, all samples were modified by 1% biopolymer and cured
800
at room temperature for 14 days. The fat cells may be placed in soil
Compressive Strength (kPa)

700
and adhesive particles, in consequence, the biopolymer cannot
600 directly interact with sand particulates. Thus, it should be noted
500 that high-quality biopolymer-soil mixture would be made by the
400 skim milk.
300
200 3.2. Direct shear test
100
0 Shear strength is an important parameter in various geotechni-
25 40 60 80 120 cal engineering aspects such as retaining walls, slope stability,
Temperature (C) design of foundations and road pavements. Internal friction angle
Casein Treated Sand Sodium Caseinate Treated Sand (Ø0 ) and cohesion (c0 ) are commonly called the shear strength
parameters in terms of effective stress. Ø0 and c0 of untreated and
Fig. 7. Effect of curing temperature on compressive strength of casein and sodium
caseinate treated soil.
Table 4
The influence of fat amount of milk on biopolymers treated soil properties.
compared with 60 °C, the compressive strength fell to a low of
Fat proportion (%) Casein treated soil Sodium caseinate treated
approximately 30% and 48% for sodium caseinate treated sand
strength (kPa) soil strength (kPa)
and casein treated soil, respectively. Therefore, the optimum ther-
0 596.76 635.34
mal condition for both biopolymers to produce the most resistant
3 427.64 495.18
mixture is curing at 60 °C. It is likely that heating up to 60 °C
818
treated sand with casein and sodium caseinate (0.5, 1%) are pre-
sented in Fig. 8. It should be noted that internal friction angle
and cohesion of sand without additives were 36.66° and 0.17
kPa, respectively. The results (see Fig. 8) also demonstrate that
shear strength parameters (c and Ø) of treated specimens
increased considerably after 14 days of curing. It can also be seen
that friction angle increased when the biopolymer amount was
increased from 0 to 1%. The casein treated soil demonstrated supe-
rior performance in increasing friction angle amount compared to
sodium caseinate treated soil samples. However, when 1% of
biopolymer was used to improve soil samples, the cohesion of
sodium caseinate treated sand rose up to 145.85 kPa; also casein
enhanced the cohesion of soil up to 124.35 kPa.
3.3. CBR test results
Unsoaked california bearing ratio test was performed for natu-
ral dune sand and sand mixed with 0.5% and 1% casein and sodium
caseinate cured 14 days at room temperature. The specimens for
CBR test were prepared at optimum moisture content and maxi-
mum dry density for both biopolymer samples. As it can be seen
from Fig. 9. There was a significant rise in CBR value with the
increase in casein and sodium caseinate biopolymer contents. A
comparison between values of sand treated biopolymers also
shows that sodium caseinate treated sand specimens represent a
greater strength than casein treated sand samples. For sodium
caseinate treated soil, the CBR values, when compared with natural
soil, increased by 96% for 0.5% biopolymer and 257% for 1%
biopolymer, while these amounts were 80% and 216% for 0.5%
and 1% of casein, respectively.
50
Angle of Internal Fricon
40
(degree)
30
20
10
0
0 0.5
Biopolymer Content
Casein
160
140
120
Cohesion (kPa)
100
80
60
40
20 0.17 0.17
0 ate into the soil grains, Van der Waals bonds, electrostatic interac-
0 0.5
Biopolymer Content (%)
Casein Sodium Caseinate
(b)
Fig. 8. variation of shear strength parameters of casein and sodium caseinate
treated sand.
H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821
100
90
80
70
60
CBR
50
40
30
20
10
0
0 0.2 0.4 0.6 0.8 1
Resin content (%)
Fig. 9. Variation of CBR of casein and sodium caseinate treated sand.
Furthermore, it should be noted that casein and sodium casei-
nate biopolymers can be produced from spoiled milk. Besides,
the amount of spoiled milk, which is usually discarded throughout
the world, has seen a considerable increase in the last few years.
The word ‘‘A River of White” was used by U. S. department of agri-
culture to indicate the high proportion of extra milk generated dur-
ing 2014 and 2015 [35]. In addition, 5.17% of all water annually
consumed in the world is associated with dairy products, so dump-
ing the extra milk wastes an immense amount of water [36].
3.4. Leaching test
Leaching test shows that the biopolymers treated sand samples
would be resistant against the water flow. The leaching test results
are shown in the Table 5. In this test, 2% of both casein and sodium
caseinate was used to improve the sand. As it can be seen, only
1.1% of casein added to the soil was leached from treated soil
which can be ignorable to the amount of leached sodium caseinate.
This is because casein is hydrophobic and insoluble in water
whereas sodium caseinate salt can be dissolved in water.
3.5. Microscopic observation
1 To observe the influences of casein and sodium caseinate on the
sand micro-structure, SEM images have been presented. Fig. 10a
shows the natural state of dune sands of sand hills in deserts. Lack
Sodium Caseinate of cohesion between particles, relatively rounded shape, and equal
size of grains cause the particulates to move on each other without
(a)
restriction. Dust particles also separate from particulates through
existing pores and disperse freely into the air. Fig. 10b indicates
the compacted untreated sand at optimum moisture content. The
pores were more compacted in comparison to Fig. 10a and a more
condensed mass of soil exists, but there is no special bond between
micro-particles.
It can be seen in Fig. 10c that casein biopolymer interacts
directly with soil particles. Most mechanisms demonstrate that
adhesion includes 4 stages: wetting, adsorption, curing, and
mechanical locking [37]. Because of the polymeric chains of casein,
these chains can paste soil grains to each other through different
mechanisms of adhesion. When polymeric chains of casein perme-
1
Table 5
Leaching test results.
Biopolymer content Pure soil Casein treated Sodium caseinate
sand (2%) treated sand
Leached content (%) 0 1.1 16.7
 H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821 819

Fig. 10. Scanning electron microscopy images, a) natural state of dune sand, b) compacted untreated sand, c) casein treated sand, d) sodium caseinate treated sand.

(a)

(b)

Casein

Sodium caseinate

Ionic interaction

Fig. 11. Protein-based biopolymers – dune sand interaction model a) Casein treated sand, b) Sodium caseinate treated sand.
820 H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821
tion or complex bonds between activated protein groups (phos-
phate group, amine group, amide group, and carboxylic acid) and
soil particles would take place.
It can be also observed from Fig. 10d that grain surfaces have
been coated by sodium caseinate, thereby increasing contact sur-
faces among sand particles. In fact, the entry of an alkaline into
the casein protein chains causes casein phosphate to join with
sodium and to form a complex structure. Additional sodium leads
to hydrolysis of amino acid chains so that polar groups would be
created on these chains. Thus, sodium caseinate particles were
charged and a strong adhesion among biopolymer treated sand
particles could be formed. Fig. 10d also shows that bridges have
been formed between detached particles by sodium caseinate.
Sodium caseinate treated sand showed higher adhesion values
than casein treated sand samples, this can be seen in Fig. 11. A
schematic model of dune sand improved by biopolymers is repre-
sented. The greater performance of sodium caseinate can be
explained with the presence of a higher amount of charge centers,
which influences the extent of ionic interactions with dune sand
grains. Consequently, a greater amount of secondary polar interac-
tions would be formed. In addition to polar interactions, sodium
caseinate residues promoted the entry of water, more efficient
chain mobility, physical chain entanglement, and favors the estab-
lishment of ionic interactions and hydrogen bonds among biopoly-
mer and sand particles.
3.6. Comparison between the sand treated by protein-based
biopolymers and other biopolymers
Different biopolymers have been used to improve mechanical
properties of sandy soils. A comparison among performances of
sand treated by various biopolymers has been conducted and the
1000
900
800
700
UCS (kPa)
600
500
400
300
200
100
0 the increase in curing time, although the strongest values for
Casein 1% Sodium Gellan 1% Agar 1% Xanthan 1%
Caseinate
1%
Fig. 12. Unconfined compressive strength of sand treated by biopolymers.
30
25
Curing me (day)
20
15
10
5 sodium caseinate to attach to soil particles so that the high
0
Fig. 13. Minimum curing time for the sand treated by biopolymers.
results presented in Figs. 12 and 13. As can be seen from Fig. 12,
the highest compressive strength has been obtained by using xan-
than gum to treat sand, while casein and sodium caseinate
biopolymers proposed a similar performance as gellan biopolymer.
In addition, the compressive strength of sand treated by 5% lignin
was significantly lower than 1% protein-based treated sand. Fig. 13
also shows how much time is needed for sand stabilized by differ-
ent biopolymers to gain at least 80% of their maximum unconfined
compressive strength. When Casein biopolymer, xanthan gum, and
agar gum added to the sand, it took 7 days to get their maximum
compressive strength (80%). Whereas, the curing time for the sand
treated by Sodium caseinate, beta glucan, and xanthan gum is 14
days. The longest curing time is also related to the lignin treated
sand by 28 days of curing. It can be concluded that casein and
sodium caseinate biopolymers have a good potential to use as
additives for sand improvement.
4. Conclusion
The primary aim of this study is to introduce two protein-based
biopolymers including casein and sodium caseinate in soil
improvement projects. These polymers are biodegradable and con-
fer environmentally-friendly capabilities in soil treatment applica-
tions. These biopolymers can be produced in the field from spoiled
milk which usually dumped in many countries. Consequently, a
series of laboratory tests were performed to study the role of
casein and sodium caseinate biopolymers in improving geotechni-
cal parameters of poorly graded sand. The uniaxial compression
tests were conducted to investigate the influences of biopolymer
content, curing temperature, curing time, and fat milk content on
compressive behavior of dune sand. The results showed that uniax-
ial strength of sand increased with the increase in casein and
sodium caseinate contents. The pasty shape of sodium caseinate
compared to cheesy characterization of casein helps to make a
more uniform mixture, so the sodium caseinate treated dune sand
samples reach greater compressive strength values.
Curing temperature affected unconfined compressive strength
significantly. More specifically, increasing the temperature from
25 °C to 60 °C increased the compressive strength for both biopoly-
mers treated soil samples. But higher temperatures (80 °C and 120
°C) caused protein-based biopolymer particles to decompose into
amino acids. As a result, the compressive strength was reduced.
The uniaxial compressive strength of specimens increased with
Lignin 5% casein treated sand were obtained within 7 days of curing and
within 14 days of curing for sodium caseinate. Besides, the results
show that soil specimens treated by biopolymers produced from
full-fat milk also yielded the lower compressive strength in com-
parison to the biopolymers prepared from skim milk.
Moreover, the interaction between sand and biopolymer was
studied by using scanning electron microscopy (SEM) images.
The images indicate that biopolymers provided an excellent condi-
tions to form resistant combination with soil particles. Van der
waals and electrostatic interaction or complex bonds among acti-
vated protein groups and sand particulates can be formed by using
casein as an additive to the soil. The sodium caseinate particles
with a complex structure coat the grain surfaces and charged
strength mixture would be formed by having strong bonds.
Direct shear tests were also performed on all neat/biopolymer
treated soil samples. Internal friction angle (Ø) and cohesion (c)
experienced considerable increase in soil mixed with biopolymers
compared to untreated soil.
CBR tests were performed on sand treated by casein and sodium
caseinate cured 14 days at room temperature. A significant
 H. Fatehi et al. / Construction and Building Materials 167 (2018) 813–821
increase in CBR values was observed for both casein and sodium
caseinate compared to untreated soil. CBR was 1.8 and 3.16 times
greater than natural sand for 0.5% and 1% casein, respectively.
These values were 1.96 to 3.57 times for sodium caseinate treated
soil samples.
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