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BLEACHING OF COTTON FABRIC WITH ENZYMATICALLY GENERATED

HYDROGEN PEROXIDE AND BLEACH ACTIVATOR

Conference Paper · June 2011

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 150 Years of Innovation and Research in Textile Science

BLEACHING OF COTTON FABRIC WITH

ENZYMATICALLY GENERATED HYDROGEN PEROXIDE
AND BLEACH ACTIVATOR
Nina Špička1 and Petra Forte Tavčer1
1
Department of Textiles, Faculty for Natural Sciences and Engineering, University of Ljubljana

nina.spicka@ntf.uni-lj.si; petra.forte@ntf.uni-lj.si

ABSTRACT of the most popular treatments. One very important

A process of enzymatically generated hydrogen
peroxide combined with bleaching of incorporated
bleach activator has been investigated. The
hydrogen peroxide was generated with enzymes
glucose oxidases which catalyse the oxidation of β-
D-glucose to D-glucono-δ-lactone, by utilizing
molecular oxygen as an electron acceptor with
simultaneous production of hydrogen peroxide. The
generated hydrogen peroxide was converted into
peracetic acid using tetra acetyl ethylene diamine
(TAED), which is one of the most successfully used
bleaching activators for hydrogen peroxide. Various
TAED concentrations were tested. The process took
place at acid to neutral pH and at temperature 50
°C. The generation and consumption of hydrogen
peroxide and peracetic acid were measured by
sodium thiosuplhate titrations. In addition, the
whiteness degree and degree of polymerization
were measured. The process showed a powerful
bleaching ability at mild process conditions. In the
process the cotton fibres suffer no damage.
Key words: glucose oxidases, hydrogen peroxide,
bleach activator, peracetic acid
1 INTRODUCTION
Biotechnology is one of the ways of making textile
processes ecologically friendlier. Enzymatic
desizing with amylases, scouring with pectinases,
biopolishing with cellulases, destroying hydrogen
peroxide after the bleaching with catalases are some
process i.e. enzymatic bleaching is still under
investigation [1-7]. Bleaching is one the essential
processes for producing high quality cotton fabrics.
Cotton is bleached with hydrogen peroxide (HP),
which is an ecologically undisputable substance.
However, bleaching with HP takes place in strong
alkali bathes at high temperature. Bleaching, rinsing
and neutralization require large amounts of water
and energy. Moreover, during the bleaching process
cotton fibres get damaged. In consequence, an
environment-friendly bleaching process would be
essential.
HP can be produced enzymatically with enzymes
glucose oxidases which catalyse the oxidation of β-
D-glucose to D-glucono-δ-lactone by utilizing
molecular oxygen as an electron acceptor with
simultaneous production of HP (Fig. 1). The
enzymatic production of HP takes place at pH 4.5 to
7 at temperatures around 40 °C. The generated HP
can be used for bleaching of textile fibres or other
purposes [8]. Investigations have been done on the
generation of HP using glucose derived from
enzymatically decomposed starch in the desizing
liquors [9, 10]. In such a system, the desizing waste
baths can be reused as an additional source of
glucose, resulting in lower water and chemical
consumption, as well as in lower waste water
pollution. The gluconic acid is formed in the
enzyme reaction from glucose and it can act as a
chelator for metal ions in the bleaching baths [11].

FIGURE 1.Representation of the glucose oxidases reaction [8]

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 150 Years of Innovation and Research in Textile Science

Until now the research was mostly focused in
enzymatic generation of HP and further bleaching
with HP at pH=11.5 at 90 °C. This kind of process still
exhibits disadvantages of the conventional bleaching
process with added HP. The goal of our research was
to gain a solution with the highest possible
concentration of HP and to use the generated HP for
bleaching of cotton fibres. The whole bleaching
process should run at milder conditions, i.e. acid to
neutral pH and temperature around 50 °C. Since the
HP bleaches fibres at pH 11.5 and at 90 °C, the
process at milder conditions could be feasible by
utilizing bleach activators which generate peracids in
situ during the bleaching process. To date, tetra acetyl
ethylene diamine (TAED) is one of the most
successfully used activators for HP bleaching and is
mainly used in the detergent industry. Bleaching of
cotton fibres with incorporated TAED bleach activator
has been investigated intensively during the last years
[12-17]. The reaction mechanism of TAED bleach
activator, where peracetic acid is generated, is
presented in Fig. 2.

FIGURE 2. TAED reaction mechanism

It is well known that peracids, such as peracetic acid
(PAA), show stronger oxidative bleaching ability than
HP, but there are safety problems for both storage and
use. The main advantage of bleaching with PAA
instead of HP is that a satisfactory degree of whiteness
can be obtained at 60 °C at neutral pH without the
addition of auxiliary agents. This results in lower
energy and water consumption both during bleaching
and rinsing of the fabric. Neutralisation of the fabric
after bleaching is not required, unlike bleaching with
HP, where large amounts of alkali must be removed
before dyeing. There is also no damage to the cotton
fibre when PAA is used [18]. The bleaching action of
PAA is mainly the result of epoxidation of the double
bonds present in unwanted coloured compounds [19].
In the PAA bleaching process it is necessary to
consider the mechanism and conditions governing the
three possibilities, hydrolysis (Eq. 1), decomposition
(Eq. 2 and 3) and oxidation (Eq. 4).

CH 3 COOOH  H 2 O  CH 3 COOH  H 2 O 2 (1)

0
 40 C , pH  5
2CH 3 COOOH T   2CH 3 COOH  O 2 (2)

x
CH 3 COOOH M
 CH 3 COOH  O2  other  products (3)

CH 3 COOOH  impurities  CH 3 COOH  oxidised  impurities (4)

2 EXPERIMENTAL genetically modified Aspergillus niger. Glucose from

Merck was added in pure form.
Materials
Generation of HP and bleaching with TAED bleach
Desized and scoured cotton fabric, 102 gm-1, was activator
obtained from Tekstina, Slovenia.
GC 199 (1500 titr.U/ml) enzymes from Solutions with various concentrations of glucose (10-30
Genencor were used for the generation of g l-1) and enzyme GC 199 (5-15 ml l-1) were prepared.
hydrogen peroxide. These enzymes are highly
purified glucose oxidase enzyme derived from a

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 150 Years of Innovation and Research in Textile Science

The demineralised water at pH 4.6 was used in

all processes. For the bleaching process with
TAED the pH was set from 4.6 to 7.5 with
addition of 2M Na2CO3. All treatments were
performed in a laboratory dyeing machine DL-
6000 Plus from Starlet at temperature 50 °C. The
fabric to liquor ration was 1:50. FIGURE 4. Procedure used in this study for HP generation and
bleaching (A-enzyme, B-glucose, C-TAED bleach activator)

Methods

FIGURE 3. Procedure used in this study for HP generation
and bleaching (A-enzyme, B-glucose)
Concentration of reactants
The rate of PAA and HP decomposition and
consumption in the bleaching solution was monitored
by iodometric titration. To determine PAA and HP
concentration the following method, developed in the
Laboratory for Peroxide Chemistry in the Faculty of
Chemistry and Chemical Technology, University of
Ljubljana was used.
PAA and HP react with potassium iodide as indicated in
Eq. 5 and 6, respectively. The iodine released then
reacts with sodium thiosulphate as in Eq. 7.

2 KI  CH 3 COOOH  CH 3 COOH  I 2  2CH 3 COOK  H 2 O (5)

2KI  H2O2  H2 SO4  I 2  K2 SO4  H2O (6)

I 2  2 Na 2 S 2 O3  2 NaI  Na 2 S 4 O6 (7)

To determine the concentration of PAA (cPAA),
titration with sodium thiosulphate was
performed in ice at 0 °C since HP, which is also
present in the solution, reacts only very slowly
with potassium iodide at this temperature. To
determine the concentration of HP (cHP) a
titration with sodium thiosulphate was carried
out at room temperature. Since both PAA and
HP react with potassium iodide at room
temperature, this titration provided the total
concentration of the two oxidants (cOX). The
concentrations of PAA and total oxidant were
calculated using Eq. 8. The difference between
the total oxidant and the PAA concentration gave
the concentration of HP (Eq. 9):
Vtitr  0.01
c PAA,OX 
Vs  2
Where Vtitr is the volume of 0.01 N sodium thiosuphate
solution consumed in the titration and Vs is the volume
of the sample.
pH
All pH measurements were made using an Iskra
MA5740 pH meter, calibrated at two points.
Whiteness
The degree of whiteness was measured on a remission
spectrophotometer (Datacolor Spectroflash SF600 Plus)
using the CIE method, according to EN ISO 105-
J02:1997(E).
Degree of polymerization (DP)
The DP was measured to determine the fibre damage
with the viscosimetric method in cuoxam.
(8)

c HP  cOX  c PAA (9)

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 150 Years of Innovation and Research in Textile Science

3 RESULTS AND DISCUSSION At first various solutions were prepared to determine the
optimal conditions for enzymatic generation of HP
Enzymatic generation of HP with glucose (Tab. I).
oxidases

TABLE I. Solution composition and conditions

Conditions S1 S2 S3 S4 S5 S6
c glucose[gl-1] 15 10 15 20 30 15
c GOD [mll-1] 5 10 10 10 10 15
pH 4.6 4.6 4.6 4.6 4.6 4.6
T [°C] 50 50 50 50 50 50
c H2O2max
0,023 0,023 0,025 0,025 0,025 0,027
[moll-1] *
*t = 180 min

FIG. 5 shows the concentration of hydrogen
peroxide generated by GC 199 at 50 °C in time
interval from 0 to 360 minutes. The
concentration of hydrogen peroxide rises at its
fastest during first 120 minutes. The
concentration of HP reaches a maximum after
180 minutes and stays constant. The reason for
the constant concentration of HP after 180
minutes is a result of consumed oxygen present
in the closed beakers of the laboratory dyeing
machine. The glucose oxidases utilize molecular
oxygen in converting glucose to D-glucono-δ-
lactone and simultaneously produce HP. Without the
present molecular oxygen the reaction stops. The
concentration of generated HP was higher when higher
concentrations of enzyme and glucose were used. On
the other hand, it seems that higher concentrations of
glucose compared to enzyme can hinder the enzymatic
generation of HP (solution containing 10 ml l-1 of
enzyme and 30 g l-1 of glucose). The highest
concentration of generated HP was measured at the
solution containing 15 ml l-1 of enzyme and 15 g l-1 of
glucose where after 180 minutes 0.027 mol l-1 of HP
was generated.

FIGURE 5. Concentration of generated HP with various concentrations of enzyme GC 199 (GOD)

and various concentrations of glucose at 50 °C and pH 4.6

Bleaching with addition of TAED bleach activator
In the second step four solutions with 15 ml l-1 of
enzyme and 15 g l-1 of glucose were prepared and
after 180 minutes various concentrations of TAED
bleach activators were added to the solutions (Tab. II).
The pH was set from 4.6 to 7.5, while the temperature
stayed at 50 °C. To study the effect of various
concentrations of TAED bleach activator on the
consumption of the bleaching agents, the
concentrations of PAA and HP were measured. The
results for the four used concentrations of TAED are
illustrated in Fig. 6. The lowest consumption of HP
with time took place when the lowest TAED
concentration was used (1,5407 gl-1) and was more
rapid with higher TAED concentrations. Commonly,
the generation and consumption of peracetic acid was
also higher when higher TAED concentrations were
used. When 1,5407 g l-1 of TAED was used around
0.0035 mol l-1 of PAA was generated. At higher
TAED concetrations the generated concentration of
PAA was very similar (0,007 mol l-1). According to
this, the consumption of HP and PAA at higher TAED
concentrations could possibly be ascribed to their
spontaneous decomposition.

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 150 Years of Innovation and Research in Textile Science

TABLE II. Concentrations of TAED bleach activator (the molar ratios were calculated according to the maximum generated H2O2
concentration)
H2O2 : TAED mol ratio 4:1 2:1 1:1 1:2
c TAED [mol l-1] 0,00675 0,0135 0,027 0,054
c TAED [g l-1] 1,5407 3,0814 6,1628 12,3255
pH 7.5 7.5 7.5 7.5
T [°C] 50 50 50 50

FIGURE 6 and 7. Concentration of HP (left) and PAA (right) after added TAED bleach activator at T=50 °C and pH=7.5

Whiteness degree and degree of polymerisation
The whiteness values (W) and the degree of
polymerization (DP) are shown in Fig. 8. Before
bleaching the whiteness value of the raw fabric was
18.11. The whiteness value of the fabric treated with
enzymatically generated HP at pH 7.5 and 50 °C is
only 31.33. At the same treatment conditions when
TAED bleach activator is incorporated a significantly
higher whiteness can be achieved, i.e. from 60 to 70.
When TAED concentrations above 3.0814 g l-1 were
used the whiteness degree remained similar. The DP
values after bleaching were lower, but the decrease
was small. There is hardly any damage to the fabric
compared with unbleached raw fabric.

FIGURE 8. Whiteness degree of samples treated with TAED bleach activator

4 CONCLUSIONS the bleaching solution. The rate of enzymatically

Combined process of enzymatically generated HP and
bleaching with TAED has been studied as an
alternative to classical bleaching with HP. Various
concentrations of enzyme, glucose and TAED bleach
activator were investigated in terms of the rate of
generation of HP and consumption of HP and PAA in
generated HP increased to a certain level in terms of
present molecular oxygen in the solution and when
higher concentrations of enzyme and glucose units
were used. The greatest generation of HP took place
in the solution containing 15 ml l-1 of enzyme and 15
g l-1 of glucose. During the bleaching process the
consumption of HP and PAA was more rapid at higher

AUTEX 2011 Conference – 8 – 10th June 2011 – Mu1house – France 184

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150 Years of Innovation and Research in Textile Science
TAED concentrations. The degree of whiteness
increased with higher concentrations of TAED bleach
activator, but was very similar. Therefore, the
consumption of HP and PAA at higher TAED
concentrations could be mainly ascribed to
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AUTEX 2011 Conference – 8 – 10th June 2011 – Mu1house – France
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185