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Preparation of super-low Protein Natural Rubber

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 ROHSTOFFE UND ANWENDUNGEN
RAW MATERIALS AND APPLICATIONS
Removal of proteins ⋅ Natural rubber
latex ⋅ Urea ⋅ Polar solvent ⋅ Nitrogen
content
An attempt was made to remove all
proteins from natural rubber in the la-
tex stage by both denaturation of the
proteins and removal of lipids, which
provided free space between them for
fast detachment of the proteins. The re-
moval of proteins was carried out by in-
cubation of natural rubber latex with
surfactant, denaturing agent and polar
organic solvent, followed by centrifuga-
tion. The nitrogen content of the resul-
ting rubbers was analyzed by the Kjeh-
dahl method. The nitrogen content de-
pended on the amount of urea, polar
organic solvent concentration, amount
of metal ions in the water and gel con-
tent. The green strength of the protein
free natural rubber was slightly reduced
compared to untreated natural rubber.
Herstellung von Naturkaut-
schuk mit niedrigem Protein-
gehalt
Entfernung von Proteinen ⋅ Naturkaut-
schuk-Latex ⋅ Harnstoff ⋅ polares
Lösungsmittel ⋅ Stickstoffgehalt
Es wurde der Versuch unternommen al-
le Proteine aus Naturkautschuk- Latex
sowohl durch Denaturierung der Prote-
ine als auch durch Entfernung von Lipi-
den, die freien Raum für eine schnelle
Ablösung der Proteine bereitstellen, zu
entfernen. Die Entfernung der Proteine
wurde durch Inkubation von Naturkaut-
schuk-Latex mit Tensiden, Denaturie-
rungsmittel und polaren organischen
Lösungsmittels, gefolgt von Zentrifugie-
ren durchgeführt. Der Stickstoffgehalt
des resultierenden Kautschuks wurde
durch die Methode nach analysiert. Der
Stickstoffgehalt war abhängig von der
Menge an Harnstoff, an polaren organi-
schen Lösungsmitteln, an Metallionen
im Wasser und vom Gelgehalt. Die Fes-
tigkeit des grünen proteinfreien Natur-
kautschuks war im Vergleich zu unbe-
handelten Naturkautschuk (FLNR) leicht
reduziert.
Figures and Tables:
By a kind approval of the authors.
46 KGK · 4 2015
Preparation of super-low Protein
Natural Rubber
Introduction
Removal of proteins from natural rubber
latex is one of the key technologies in
natural rubber science and technology,
since natural rubber contains about 6
w/w% non-rubber components inclu-
ding proteins, lipids and other content,
which results in both side reactions du-
ring modification [1-2] and latex allergy
in users of end products [3-7]. The remo-
val of the proteins may be carried out in
the latex stage because the non-rubber
components are known to exist on the
surface of the rubber particles as a mo-
nolayer; for instance, after coagulation
of natural rubber latex, few proteins may
be removed from natural rubber. The
proteins on the surface of the rubber
particles may be efficiently detached in
the latex stage by controlling chemical
and physical interactions using denatu-
rant.
The interaction between the proteins
and the lipids on the surface of the rub-
ber particles is described to be integral
membrane proteins and phospholipid
monolayer because the hydrophobic side
chains of the integral proteins may inter-
act with hydrophobic tails of the lipids
thereby holding the proteins onto the
surface of the rubber [8-11], as shown in
Figure 1. In the previous work, to remove
the proteins, the proteolytic enzyme was
used to cleave the peptide linkages of
the proteins [12] while urea was used to
change the conformation of the proteins
[13-14]; that is, only proteins were dena-
tured whereas lipids were not taken into
account. As the lipids interact extensi-
vely with the proteins on the surface the
rubber particles, therefore, it is impor-
tant to consider the removal of the lipids,
as well. By removing the lipids on the
surface, the proteins may be removed
effectively.
Over the last three decades, several
approaches to remove the proteins from
natural rubber latex were performed.
The most conventional method was de-
proteinization with a proteolytic enzyme
i.e. an alkaline protease [15-17]. The en-
zymatic treatment involved cleavage of
the peptide linkages of the proteins to
produce oligopeptides or low molecular
weight protein fragments. These are so-
luble in water and therefore easily remo-
ved by washing the latex. The deproteini-
zation of the latex with the enzyme re-
duced the nitrogen content to 0.1-0.02
w/w% from ca. 0.40 w/w% This implies
that the proteins still remained after the
enzymatic treatment. The small frag-
ments of the proteins on the surface of
the rubber particles may not be comple-
tely removed after assimilation of the
proteins. Therefore, urea as a denaturing
agent was applied to denature the prote-
ins [13, 18-19], which involved the physi-
cal interaction that may change the con-
formation of the proteins to result in vo-
lume contraction. It was reported that
the nitrogen content was markedly redu-
ced to 0.02-0.005 wt%. A further challen-
ge to remove all proteins from natural
rubber latex is pronounced, since the re-
sulting latex is in high demand. Most re-
cently, protein free natural rubber latex
was prepared by incorporating the dena-
turing agent and polar organic solvent
into the natural rubber latex [20]. By
combining urea and acetone, a good sol-
vent for lipids, may provide free space
between the proteins and the lipids and
assist in fast detachment of the proteins.
The resulting nitrogen content obtained
was 0.00 w/w%, which implies no prote-
ins were attached to the surface of natu-
ral rubber particles.
Authors
Nurul Hayati Yusof,
Seiichi Kawahara, Nagaoka,
Niigata, Yoshimasa Yamamoto,
Tokyo (Japan)
Oraphin Yamamoto, Pathumtha-
ni (Thailand)
Corresponding author:
Prof. Dr. Seiichi Kawahara
Department of Materials Science
and Technology, Faculty of Engi-
neering, Nagaoka University of
Technology, Nagaoka, Niigata
940-2188, Japan
main author e-mail: hayati@lgm.
gov.my,
E-mail: kawahara@
mst.nagaokaut.ac.jp
www.kgk-rubberpoint.de
 ROHSTOFFE UND ANWENDUNGEN
RAW MATERIALS AND APPLICATIONS

However, the reproducibility of the 1 Fig. 1: Protein-lipid in-

nitrogen content of the product was not teraction on the sur-
ascertained, since the removal of the face of rubber particle.
proteins depended on the seasons and
rubber clones [21-22]. The composition Lipid
of the non-rubber components may va-
ry, resulting in the difference in the nit- Rubber
rogen content. Hence, several factors, particle
e.g. gel content, must be investigated to Protein
prepare the protein free natural rubber,
reproducibly. This is because the gel
content is related to the nitrogen con-
tent, according to the previous work
[23-25]. Prolonged storage of natural
rubber latex may increase the gel con-
tent [26]. In addition, the amount of Removal of proteins for 30 minutes. After each centrifugation,
metal ions in water in different coun- from natural rubber latex the cream fraction was collected and re-
tries may also affect the gel content. Removal of the proteins from HANR and dispersed in the solution with SDS and
Therefore, it would be of interest to FLNR latexes were carried out by incubati- polar solvent. Finally, the SDS concentrati-
know whether gel content and amount on of the latexes with urea and polar sol- on of the resulting latexes was adjusted to
of metal ions in water (type of water) vent in the presence of SDS. The HANR and 0.1 w/w%. The film specimens were pre-
affect the nitrogen content. FLNR latexes were diluted with deionized pared by casting the latexes and serum
In the present work, an attempt was water to 30 w/w% of dry rubber content fraction onto the petri dish and then dried
made to investigate the effect of the gel (DRC) before use. Definite amounts of in the oven at 50 °C. The procedure to re-
content, the amount of metal ions in the urea, SDS and polar solvent were added move the proteins is shown in Figure 2.
water and the chemicals such as SDS and into the latexes. The latexes were then The conditions for the removal of the pro-
polar solvent concentrations and the centrifuged (ca, 10,000 g) 3 times at 15 °C teins are shown in Table 1.
amount of urea in order to prepare the
protein free natural rubber latex. The re- 2 Fig. 2: Removal of the
sulting rubbers were characterized by proteins from natural
HANR or FLNR latexes rubber latex
Kjehdahl method, swelling method, fatty
acid ester content by FTIR spectroscopy, + 1,2w/w% SDS
transmission electron microscopy (TEM) + 0.025, 2.5w/w% solvent
and tensile strength measurement. The
+ 0.1 – 0.5w/w% urea
first approach was to use high ammonia
natural rubber (HANR) latex as a source 1 hour incubation
to determine the optimum condition.
The second approach was to apply the
optimum condition to fresh field natural Purified natural rubber latex First Centrifugation
rubber (FLNR) latex. After the removal of Collect cream fraction
the proteins, they were referred to as
+ 0.5 – 1w/w% SDS
protein-removed HANR or FLNR, respec-
tively. + 0.025, 2.5w/w% solvent
1 hour incubation
Experimental
High ammonia natural rubber (HANR)
Purified natural rubber latex Second Centrifugation
latex was purchased from Lee Rubber
Company (Malaysia) and Golden Hope Collect cream fraction
Plantation (Malaysia). Fresh field natu-
+ 0.1w/w% SDS
ral rubber (FLNR) latex was collected in
Malaysia and Vietnam. Urea used in this + 0.025, 2.5w/w% solvent
study was purchased from Acros, sodi- 1 hour incubation
um dodecyl sulfate (SDS) was from Sig-
ma Aldrich and solvents such as acetone
(A), ethanol (E), methanol (M) and tolue- Purified natural rubber latex Third Centrifugation
ne were from Merck. Chemicals for
Collect cream fraction
Kjehdahl method, i. e. potassium sulfa-
te, copper sulfate, selenium, sodium hy- + 0.1w/w% SDS
droxide, concentric sulfuric acid, boric
acid, 0.005 M H2SO4 and methyl red so- Protein-removed HANR or FLNR latexes
lution were used as received from
Merck.

www.kgk-rubberpoint.de KGK · 4 2015 47

 ROHSTOFFE UND ANWENDUNGEN
RAW MATERIALS AND APPLICATIONS

1 Tab. 1 Conditions for removal of the proteins 2 Tab. 2 Gel content of HANR and
Acetone (%) Methanol Ethanol (%) SDS (%) Urea (%) FLNR latexes (starting materials)
(%) Latex Gel content (wt%)
A-0.025 0.025 - - 1, 2 0.1, 0.3, 0.5 HANR-1 64.32
A-2.5 2.5 - - 1, 2 0.1, 0.3, 0.5 HANR-2 58.33
M-0.025 - 0.025 - 1, 2 0.1, 0.3 HANR-3 56.55
M-2.5 - 2.5 - 1, 2 0.1, 0.3 HANR-4 46.96
E-0.025 - - 0.025 1, 2 0.1, 0.3 FLNR-1 37.53
E-2.5 - - 2.5 1, 2 0.1, 0.3 FLNR-2 11.96

Characterizations
Measurement of the nitrogen content of
the film specimens was carried out by the
Kjehdahl method as described in RRIM
Test Method B7 [27]. The film specimens
weighing about 0.1 g were digested with
about 2.5 ml concentrated H2SO4 and 0.65
g catalyst mixture (potassium sulfate:
copper sulfate: selenium with a 15:2:1
weight ratio). The resulting solutions were
mixed with 10 ml sodium hydroxide solu-
tion (67 w/v%) followed by steam distilla-
tion. The distillates, added with 2 w/v%
boric acid, were titrated with 0.005 M
H2SO4 using methyl red as an indicator.
Each sample has three replicates and
measurement of nitrogen content was
carried out three times for each replicates.
The amount of metal ions present in
the water used for the removal of the
proteins was assessed by element analy-
sis. The water was directly analyzed with
inductively coupled plasma optimal
emission spectroscopy, ICP-OES (Perki-
nElmer, OptimaTM 4300DV).
The gel content was determined by
the swelling method. A rubber sample
weighing 0.5 g was dissolved in 100 g
dried toluene, and the resulting solution
was placed in the dark at room tempera-
ture for one week. The toluene-insoluble
(gel) fraction was separated by centrifu-
gation followed by filtration. The gel
fraction was then dried in a vacuum oven
for one week. The gel content of the
HANR and FLNR were determined as
shown in Table 2 in order to study the
effect of gel content on nitrogen content
as discussed in results and discussion
section. Measurement of gel content was
carried out three times for each sample.
The gel content is calculated using the
following equation:
Gel content, % =  (1)
Weight of dried gel fraction, g  x 100
Weight of rubber, g
The fatty acid ester content in the samp-
les was determined by Fourier transform
3 Tab. 3 Effect of the type of water on the
nitrogen content of the films prepared from
protein-removed HANR latex
Nitrogen content (w/w%)
Japan Malaysia
A-0.025 0.023 0.013
A-2.5 0.014 0.010
4 Tab. 4 Amount of metal ions in distilled
water used from Japan and Malaysia
Element (ppm) Japan Malaysia
Ca 0.637 - Malaysia-water). In the experiments, two
K 0.213 0.184
Mg 0.174 0.003
Na 1.050 0.271
Fe - -
infrared spectroscopy (Jasco FT/IR4000).
A calibration curve was obtained for a
series of mixtures of methyl stearate and
synthetic cis 1,4 polyisoprene with R2 =
0.99. The fatty acid ester content per
weight of rubber was estimated by peak
intensity ratios at 1740cm-1 (C=O) to
1664cm-1 (C=C).
Tensile testing was carried out at
room temperature using an Instron uni-
versal tensile tester (Instron model
3365), according to Japanese Industrial
Standard K 6251 (JIS K6251). The film
specimens with a thickness of about
1 mm were cut with dumbbell-shaped
Type 7. The test piece was stretched with
a crosshead speed of 200 mm/min at
room temperature. The test was carried
out 5 times for each sample. The data
were plotted in stress-strain curve.
The morphology of the resulting rub-
bers was observed with a transmission
electron microscope, TEM (Jeol, JEM-
2100), at an accelerating voltage of
200 kV. The ultrathin sections of the
samples were prepared by cryo-microto-
me (Ultracut N, Reichert-Nissei FC S) at a
temperature lower than the glass tran-
sition temperature of the samples. The
ultrathin sections with the thickness of
about 100 nm were stained with phos-
photungstic acid (PTA) at room tempe-
rature for 10 min.
Results and discussion
Effect of amount of metal ions in water
Table 3 shows the effect of the type of
water on the nitrogen content of the
films prepared from the protein-remo-
ved HANR latex. Distilled waters from
Japan and Malaysia were used respec-
tively (abbreviated to Japan-water and
conditions were adopted: HANR latex
treated with 1 w/w% SDS, 0.1 w/w%
urea, 0.025 w/w% acetone (A-0.025) and
HANR latex treated with 1 w/w% SDS,
0.1 w/w% urea, 2.5 w/w% acetone (A-
2.5). It was found that the experiments
with Japan-water gave higher nitrogen
contents: 0.023 w/w% for A-0.025 and
0.014 w/w% for A-2.5. By comparison,
results with Malaysia-water were 0.013
w/w% for A-0.025 and 0.010 w/w% for
A-2.5. In addition, the 2.5 w/w% acetone
gave the lowest nitrogen content. This
demonstrates that the role of solvent in
assisting the removal of proteins from
natural rubber latex, as mentioned in in-
troduction section. According to the me-
tal ions analysis in Table 4, the metal ions
content in Japan-water was found to be
higher than in Malaysia-water. This indi-
cates the higher nitrogen content is due
to higher metal ions content. According
to the previous work [28-29], the magne-
sium (Mg) ions were found to form ionic
linkages between the natural rubber
chain and phospholipid groups, which
contribute to gel formation, especially, in
HANR-latex. Thus, the presence of the
Mg ions may positively contribute to the
formation of the ionic linkages at the
phospholipid terminal units and cause a
rise in the gel content and also nitrogen
content. This demonstrates that the type
of water such as soft (Malaysia) and hard
(Japan) waters may influence the nitro-
gen content. Hence, the removal of the
proteins from natural rubber latex must

48 KGK · 4 2015 www.kgk-rubberpoint.de

 ROHSTOFFE UND ANWENDUNGEN
RAW MATERIALS AND APPLICATIONS

5 Tab. 5 Effect of the SDS concentration on 3

the nitrogen content of the films prepared 0.03
from the protein-removed HANR latex
SDS concentration 0.025
Control 1 w/w% 2 w/w%
HANR 0.338 - -

Nitrogen content (%)

0.02
A-0.025 - 0.019 0.018
A-2.5 - 0.019 0.019 0.015 A-0.025
M-0.025 - 0.020 0.017
0.01 A-2.5
M-2.5 - 0.017 0.016
M-2.5
E-0.025 - 0.017 0.018 0.005
E-2.5 - 0.018 0.017 E-2.5
0
be carried out in soft water, which con- 0 0.1 0.2 0.3 0.4 0.5 0.6
tains low level of metal ions in order to
obtain low nitrogen content. In the sub- Urea concentration (%)
sequent section, the work was carried
Fig. 3: Effect of the amount of urea on the nitrogen content of the films prepared from
out with soft water.
the protein-removed HANR latex.

Effect of SDS concentration

on nitrogen content
Table 5 shows the effect of the SDS con-
centration on the nitrogen content of the
films prepared from the protein-remo-
ved HANR latex. The SDS concentration
with three types of polar solvents, i.e
acetone (A), ethanol (E) and methanol
(M) were used in this study. After the re-
moval of the proteins, the nitrogen con-
tent was markedly reduced to 0.02 w/w%
from 0.34 w/w%. This demonstrates that
SDS contributes to the removal of the
proteins with urea and polar solvents.
The nitrogen content of the films prepa-
red with 1 w/w% SDS was similar to that
of the nitrogen content of the films pre-
pared with 2 w/w% SDS. This may be ex-
plained due to the high amount of sur-
factant, which may form more micelles
in the latex. The stable micelles of SDS
may exist independently in the latex and
it may not contribute to the removal of
proteins [30], resulting in no significant
difference in nitrogen content. In additi-
on, films prepared with 2 w/w% SDS
were non-uniform whereas films prepa-
gen content of each of films prepared
with 0.1 w/w% urea was similar, even
when they were treated with the diffe-
rent solvents. The nitrogen content incre-
ased from 0.019 w/w% to 0.028 w/w%
when the amount of urea was increased
from 0.1 w/w% to 0.5 w/w%. The nitro-
gen content of the films prepared with
0.5 w/w% urea were the highest among
the samples. It is possible to postulate
that residual urea leads to higher nitro-
gen content. In this respect, if we had
added a certain amount of urea to the
latexes, the nitrogen content of the se-
rum fraction would increase. This is be-
cause the serum fraction of the treated
latexes contains the non-rubber compo-
nents and residual urea. In order to prove
the postulation, the nitrogen content of
the serum fraction was determined as
shown in Figure 4. The nitrogen content
4
2.6
Serum - Nitrogen content (%)
of the films prepared from serum frac-
tion was almost independent of the
amount of urea, despite the nitrogen
content of the film prepared from the
latexes depended on it (Figure 3). This
demonstrates that some of the residual
urea may remain in the cream fraction of
protein-removed HANR latex and result
in higher nitrogen content of these films.
Based on this finding, the suitable
amount of urea was determined to be
0.1 w/w%. Furthermore, it may be con-
cluded that the optimum condition for
the removal of proteins from HANR latex
is 1.0 w/w% SDS, 0.1 w/w% urea and 2.5
w/w% acetone.
Effect of gel content on nitrogen content
Figure 5 shows the effect of the gel con-
tent of HANR and FLNR latexes (starting
materials) on the nitrogen content of the

red with 1 w/w% SDS were uniform. This

may be explained due to the negative 2.4
effect of SDS on the formation of a uni-
form film. Because of this, in the subse-
quent works, the SDS concentration was A-0.025
determined to be 1 w/w%. 2.2 A-2.5

Effect of amount of urea M-2.5

on nitrogen content E-2.5
Figure 3 shows the effect of the amount 2
of urea on the nitrogen content of the 0 0.1 0.2 0.3 0.4 0.5 0.6
films prepared from the protein-remo-
Urea concentration (%)
ved HANR latex. The respective amount
of urea, thus added, was 0.1, 0.3 and 0.5
Fig. 4: Effect of urea on nitrogen content of films prepared from the serum fraction.
w/w%, for each polar solvent. The nitro-

www.kgk-rubberpoint.de KGK · 4 2015 49

 ROHSTOFFE UND ANWENDUNGEN
RAW MATERIALS AND APPLICATIONS
5 after removal of the proteins. The values
0.025
0.02
Nitrogen content (%)
0.015
0.01
0.005
0 a strain of 6 for HANR film was quite si-
0 10
Fig. 5: Effect of the gel content of HANR and FLNR latexes on the nitrogen content of the
films prepared from the protein-removed HANR and FLNR latexes.
films prepared from the protein-remo-
ved HANR and FLNR latexes. The starting
materials contained different amount of
the gel fractions, as shown in Table 2. The
nitrogen content was dependent on the
gel content. The higher the gel content,
the higher the nitrogen content. The
highest gel content of HANR latex was
64 w/w% and resulted in the highest nit-
rogen content, 0.02 w/w%. The lowest
gel content of FLNR latex was 12 w/w%
and resulted in a nitrogen content of
0.00 w/w% (A2.5-FLNR). Thus, the remo-
val of the proteins must be carried out
with fresh field natural rubber latex as
the starting material in order to prepare
protein free natural rubber latex.
Figure 6 shows the TEM images for
FLNR, HANR and A2.5-FLNR films. The
bright areas represent the rubber partic-
les and the gloomy areas represent the
non-rubber components. In FLNR and
HANR films, the bright areas of the rub-
ber particles were well dispersed within
the gloomy areas of non-rubber compo-
nents. For FLNR film, the gloomy areas
were more accentuated compared to
6 in the latex stage. This suggests that the
FLNR Film
0.47 w/w%
Fig. 6: TEM images for FLNR, HANR and A2.5-FLNR films.
50 KGK · 4 2015
20 30 40 50 60 70
Gel content (%)
HANR film. In the case of A2.5-FLNR film,
the gloomy areas were disappeared. This
demonstrates the non-rubber compo-
nents were effectively removed, by the
incubation of FLNR latex with 1 w/w%
SDS, 0.1 w/w% urea and 2.5 w/w% ace-
tone. These images were also consistent
with the nitrogen content, where the ni-
trogen content of FLNR film is the high-
est, 0.47 w/w% compared to 0.34 w/w%
and 0.00 w/w% in the HANR and A2.5-
FLNR films respectively. Based on the
observations, the non-rubber compo-
nents play a significant role in the forma-
tion of the nanomatrix, which consists of
natural rubber particles and matrix of
non-rubber components such as proteins
and lipids.
Green strength of natural rubber
The stress-strain curves for FLNR, A2.5-
FLNR, HANR and A2.5-HANR films are
shown in Figure 7. The value of the stress
at break of FLNR film was 6.86 MPa,
which was the highest among the samp-
les. The value of the stress at break of the
A2.5-FLNR film was reduced to 5.37 MPa,
HANR Film A2.5-FLNR Film
0.34 w/w% 0.00 w/w%
of the stress at break of HANR and A2.5-
HANR films were 5.48 MPa and 4.28
MPa, respectively. The values of the stress
at break decreased in the order of FLNR,
HANR, A2.5-FLNR and A-2.5-HANR. This
demonstrates that the value of the stress
at break depends on proteins; whereby
the value decreases after the removal of
the proteins.
The stress at a strain of 6 for FLNR film
was abruptly increased compared to
A2.5-FLNR film. In contrast, the stress at
milar to A-2.5-HANR film. According to
the previous work, the abrupt increase in
stress against strain of the untreated
natural rubber was explained to be due
to the strain induced crystallization of
natural rubber [31-32]. Likewise, the lo-
wer values of stress at a strain of 6 for
A2.5FLNR film can be explained by lower
strain induced crystallizability of the rub-
ber. These findings were consistent with
the fatty acid ester content as shown in
Table 6, whereby the fatty acid ester con-
tent of FLNR film was higher than A2.5-
FLNR film. In the case of HANR and A-2.5-
HANR films, no significant difference
was observed in the stress at a strain of
6. This was explained due to a similar
strain induced crystallizability of the rub-
ber before and after the removal of the
proteins, in which the crystallization be-
havior was associated with the amount
of linked fatty acids.
Conclusions
The optimal condition for the removal of
the proteins from natural rubber latex
was achieved by the incubation of fresh
field latex, i.e. low gel content latex, and
soft water, i.e. low metal ion water, with
the presence of 1 w/w% SDS, 0.1 w/w%
urea and 2.5 w/w% acetone. The nitro-
gen content was markedly reduced to
0.00 w/w%, which indicated that the
proteins were totally removed from the
surface of natural rubber latex particles
proteins are attached to the surface of
natural rubber particles are formed by
physical interaction, not chemical inter-
action. The nanomatrix of the non-rub-
ber components had disappeared after
the removal of the proteins, resulting in
0.00 w/w% nitrogen content. The green
strength of protein free natural rubber
(A2.5-FLNR) was slightly reduced when
compared to untreated natural rubber
(FLNR), suggesting the removal of the
proteins under the optimal condition sig-
www.kgk-rubberpoint.de
 ROHSTOFFE UND ANWENDUNGEN
RAW MATERIALS AND APPLICATIONS

7
8
1.6
A2.5-FLNR film 1.4
7
FLNR film
1.2
6 A2.5-HANR film
HANR Film 1

Stress, σ
5 0.8
Stress, σ

4 0.6
0.4
3
0.2
2 0
0 2 4 6
1 Strain, γ

0
0 5 10 15 20
Strain, γ

Fig. 7: Stress-strain curves for (a) FLNR film, (b) A2.5-FLNR film, (c) HANR film, (d) A2.5-HANR film.

nificantly affects the protein linkages but
only affects some of the phospholipids
linkages. Hence, the presence of the ori-
gin phospholipids i.e. fatty acids ester
group in natural rubber plays an impor-
tant role to control the excellent proper-
ties of natural rubber.
Acknowledgement
This work was supported in part by a
Grantin- Aid (26620176) for Challenging
Exploratory Research and Grant-in-Aid
(25288098) for Scientific Research (B)
from Japan Society for the Promotion of
Science and JST-JICA SATREPS.
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