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Section-IV: Application
Section-IV: Application 131
Section-IV
Application of Reactive dyes
Study of dyeing properties
The present section deals with the study of applicability of reactive dyes
synthesized in section-II on cotton, wool and silk. These dyes have been applied to the
said fabrics by exhaust dyeing procedure. The exhaustion and fixation studies have
been carried out. The fastness properties of dyed patterns have also been studied.
The reactive dyes belong to the fibre-reactive class of colorants and have the
unique property to combine chemically with cellulose to form a stable chemical
linkage in presence of alkali. The main reasons for such growth in the market share of
reactive dye are a wider range of shades, higher brilliance, higher wet fastness and
that they can be applied by simple methods at an economical cost. Triazinyl dyes i.e.
dye containing cyanuric chloride residues, are supposed to react with cellulose,
forming a covalent bond between the dye and cellulose.
The introduction of reactive dyes by ICI in 1956 is an important landmark in
the history of synthetic dyes[1]
. The procion dyes, on account of the sulphonic acid
group in their molecules, are readily soluble in water. In neutral solution, they have
substantivity, towards cellulose similar to that of very ‘low affinity direct dyes’,
a wash fastness of low order, and they exhaust better in the presence of an electrolyte.
In the neutral solution, physical adsorption and possibly some hydrogen bonding take
place but covalent bond formation occurs only when alkali has been added to the dye
bath. The fact that, after the addition of alkali, chemical union takes place is based
upon following facts[2]
.
(i) Before alkali treatment the color can be washed out with repeated extractions
with neutral water,
(ii) After alkali treatment, it is fast even to boiling soap solution,
Section-IV: Application
Section-IV: Application 132
(iii) The color can be stripped from cellulosic materials dyed with direct dyes,
azoic or vat dyes by boiling with pyridine, o-chlorophenol or chloroform, but
this is not the case with procion dyes fixed by the action of alkali. The
conclusion, therefore, is that a binding force of different nature is operating,
(iv) An azo dye can be reduced giving a colorless mixture of two primary amines.
If the reactivity of the dye is increased considerably, the rate of reaction with
the fibre increases. Therefore, the dyeing can be carried out in a short time. However,
in this case the rate of hydrolysis of the dye also increases, leading to deactivation of a
part of the dyes. As the temperature increases the rate of this hydrolysis increases and
for this reason, when dissolving the dyestuff and during its application in the dye bath,
the temperature is not allowed to rise above 40ºC.
However, many problems arise from the use of reactive dye (e.g.):
(a) High sodium chloride or sodium sulphate concentration (up to 100 g/lit.) is
required, which causes environmental problem.
(b) Dye-free reaction is relatively inefficient, which in some cases, results in
50-60% of the applied dye being co-valently bonded to substrate.
(c) To remove unfixed dye, time and energy consuming and expensive washing
off procedures are required.
(d) Unfixed reactive dye and/or hydrolyzed dye may cause an environmental
hazard.
Cellulose dyed with procion azo dye is reduced; one component of the dye
molecule remains anchored to the fibre. The resulting fibre is colorless. If this is
diazotised and coupled with suitable coupling component the color is restored.
This operation involving reduction, diazotization and coupling can be repeated
producing similar results.
Section-IV: Application
Section-IV: Application 133
Concurrently with the fixation of the dye on the cellulose an undesirable
reaction takes place between the chlorine atoms in cyanuric chloride group and water,
yielding either the mono or di-hydroxy compound.
Application of reactive dyes to silk fabrics
Recently there has been increased interest in the application of reactive dyes
on silk. Silk is the most superior natural fibre because of its smooth, soft luster and
elastic characteristics. It combines strength with lightness, durability with beauty and
cleanliness with luster[3]
. However, because of its long continuous form, silk is not
bulky, a characteristic considered very desirable, at present, for textile fibres and
fabrics.
Experimental
This section deals with the application of the reactive dyes on silk. This
section is divided into four subsections dealing respectively with the application of the
dye, study of exhaustion of dye bath, study of fixation of dye on the fabrics and the
study of the fastness properties of the dyed patterns.
(1) (i) Application of reactive dyes to silk fabrics
Application of reactive dyes to silk fabric was carried out by the following
procedure:
Materials and conditions for 2 % shade
Weight of the silk fabric : 2.0 gm
Amount of the dye under study : 40 mg
Acetic acid solution (10 % w/v) : 2.0 ml
Formic acid solution (40 % w/v) : 1.5 ml
MLR : 1:40
Total volume of the solution in dye bath : 80 ml
pH of the dye bath : 3 .0
Dyeing temperature : 85ºC
Time for dyeing : 90 min.
Section-IV: Application
Section-IV: Application 134
(a) Pretreatment of the fabric
Silk fabric (2.0 g) was treated with a ½ to ¾ % soap solution at near boil for
2 h keeping liquor ratio 30:1. The fabric was then removed from the bath and rinsed
several times with water squeezed, dried and conditioned for two days in atmospheric
condition and subsequently used for dyeing.
(b) Adjustment of pH of fabric
In order to study dyeing at certain pH, it is necessary to pH at the suspended
fabric and solution on forming dyebath. A weighted amount of previously treated silk
fabric was heated to 40ºC for 5 minutes in a solution which was adjusted to the
required pH 3.0 by adding diluted acetic acid solution. This treatment was continued
till pH of the solution remained fixed at pH 3.0. The silk fabric was removed and
squeezed as much as possible to free it from adhering mother liquor.
(c) Dyeing procedure
The dyestuff under study was dissolved by pasting up in cold water and then
addition of hot water and stirred well to give a clear solution. The pH of the dye bath
was adjusted to 3.0 by adding acetic acid solution and the total volume was adjusted
to 80 ml by adding required amount of distilled water. The temperature of the dye
bath was adjusted to 50ºC and the silk pattern was introduced into the dye liquor with
stirring. The temperature of the dye bath was gradually increased to 85ºC over a
period of 10 min. At this temperature formic acid was added to the dye bath to
achieve good exhaustion. The dyeing was continued for 60 minutes more. After this,
the dye liquor was decanted in to a 250 ml volumetric flask.
The dyed pattern was then washed for several times with cold water (100 ml).
The washings were transferred to the volumetric flask. The combined solution of the
dye liquor and washing was then diluted to 250 ml with water. 5 ml of this solution
was further diluted to 25 ml with water and the absorbance of this solution was
measured.
The above dyed-pattern was further treated with solution of detergent Lissapol
D (0.29 g) and sodium carbonate (0.1 g) in water (100 ml) at 50ºC for 15 min. After
washing the dyed-pattern with water, it was dried and a part of it was mounted on the
shade cards 1 and 2.
Section-IV: Application
Section-IV: Application 135
(ii) Exhaustion study of the dye bath
After diluting the liquor and washing to 250 ml, 5 ml of this solution was then
diluted to 25 ml with water. The percentage dye bath exhaustion was calculated by
measuring the absorbance of the above solution and reading the corresponding
concentration on the calibration curve. The results of the exhaustion study of each dye
for silk fibres are presented in Table-28.
Calibration study
This involved preparation of dye solution of known concentration and
measurement of absorbance at specified wavelength.
The dye under study (10 mg) was dissolved in water (50 ml) to yield a stock
solution. Four dilutions were made by diluting 0.5 ml, 1.0 ml, 1.5 ml and 2.0 ml of the
stock solution to 25 ml by adding required amount of water yielding dye solutions
containing 4.0×10-3
, 8.0×10-3
, 12.0×10-3
and 16.0×10-3
mg of dye per one ml of the
solutions were measured at various wavelengths in the range from 350 nm to 700 nm.
The position of the maximum absorbance (λmax) was ascertained from this study.
Optical densities of the above mentioned four solutions were measured at wavelength
where maximum absorbance is observed. Calibration curves were prepared by
plotting absorbance versus concentration. The relevant calibration curves for different
dyes are shown in Figures- 37, 38, 39 and 40 the data are presented in Table-26.
(iii) Fixation study of the dyed silk fabrics
This involves two steps. Construction of calibration curve and estimation of
dye which is fixed on the fabric by extraction method.
Calibration study
This involved preparation of dye solution of known concentration and
measurement of absorbance at specified wavelength.
The dye under study (10 mg) was dissolved in A.R. grade concentrated
sulphuric acid (50 ml). Using this stock solution, four other solution of lower
concentration was prepared. For this purpose 0.5 ml, 1.0 ml, 1.5 ml and 2.0 ml of the
stock solution were diluted to 25 ml by adding required amount of concentrated
sulphuric acid. The absorbance of the stock solutions was measured at various
wavelengths in the range from 350 to 700 nm. The position of the maximum
Section-IV: Application
Section-IV: Application 136
absorbance (λmax) was ascertained from this study. Optical densities of the above
mentioned four solutions were measured at wavelength where maximum absorbance
is observed. Calibration curves were prepared by plotting absorbance versus
concentration. The relevant calibration curves for different dyes are shown in Figure-
41, 42, 43 and 44 and the data are presented in Table- 27.
Estimation of fixation of dye
Dyed silk pattern (0.1 g) was placed in a corning tube and sulphuric acid (10
ml) was poured over it. This requires about 10-15 minutes stirring of fabric in
sulphuric acid at room temperature. The solution was diluted to 50 ml with sulphuric
acid. The percentage fixation of the dye was then calculated by measuring the
absorbance of this solution and reading the corresponding concentration on the
calibration curve. A solution of the same amount of undyed silk fabric in sulphuric
acid (50 ml) was used as reference solution in colorimetric estimation. The results of
the fixation study of each dye are presented in Table-28.
(iv) Study of light, wash and rubbing fastness properties of dyed silk fabrics
Study of Light Fastness
It is common practice to study light fastness properties of dyed patterns by
automatic devices known as fadeometers. Xeno-test machine is a much better
instrument for this purpose. For the present study improved tests were carried out
using daylight as a source of light. This requires use of standard dyed-pattern with
increasing light fastness properties. Such standard samples are available and are made
by the procedures according to the specifications laid down by the international
organization for the standardization (I.S.O.)[4]
. These standard samples have light
fastness properties rated from 8 to 1, the order indicating a decrease in the fastness
properties.
In actual study the dyed patterns and the eight standard dyed patterns of nearly
equal size were mounted on cards of a convenient size (~35 × 10 cm). Nearly one half
of each dyed pattern was covered with an opaque sheet of cardboard leaving the
remaining half exposed to light. The cards were then exposed to sunlight in exposure
frames inclined at an angle of 45ºC. After definite intervals of time the exposed and
unexposed portions of the test patterns were examined.
Section-IV: Application
Section-IV: Application 137
Simultaneously a similar observation was made with the standard patterns
starting with the first standard patterns. Suppose at a given time the test pattern has
not faded but the standard patterns rated at 1, 2 and 3 have faded. At the next
consecutive inspection the test pattern has distinctly faded but the standard pattern
rated as 4 has not faded. The test pattern showing such behavior is considered to have
light fastness between 3 to 6.
Grade Qualitative assessment[5]
8 Maximum fastness
7 Excellent fastness
6 Very good fastness
5 Good fastness
4 Fair fastness
3 Moderate fastness
2 Slight fastness
1 Poor fastness
Following the above procedure the light fastness properties of all the test
patterns were evaluated and assessed. The results are tabulated in Table-31.
Study of Wash Fastness
Fastness to washing is dependent on the substantivity and polarity of the dye
molecule and also on the fibre morphology. The study was carried out in the
following manner. A soap solution, which was recommended for I. S.O. test[6]
, was
prepared by dissolving soap (5 g) in distilled water (100 ml). The test dyed pattern
was then treated with 20ml of the soap solution for 45 min at 50ºC after treatment; the
test pattern was removed from soap solution bath and washed for 5-10 min in cold
running tap water. The pattern was squeezed and dried at 50ºC. Loss in depth in shade
of the pattern was assessed with grey scale. Grey scale[7]
, for alternation of color,
consisting of grade 1 to 5, defines the following:
Section-IV: Application
Section-IV: Application 138
Gray scale in light of I.S.O. recommendation
Observation Grade Qualitative assessment
Shade unaltered 5 Excellent
Very slight loss in depth or alteration 4 Very good
Appreciable loss or alteration 3 Good
Distinct loss or alteration 2 Fair
Great loss or much altered 1 Poor
By following above procedure the wash fastness properties of all the test
patterns were evaluated and assessed. The results of wash fastness study of dyes D1 to
D40 are presented in Table-31.
Study of Rubbing (crocking), Dry and Wet fastness[8]
Transfer of color ‘crocking’ from textile material is undesirable and causes
much trouble both in textile finishing and processing as well as during wear of
garments. This test is applicable to textile made from all fibres, in the form of yarns
and fabrics.
The specimens are fastened in the crockmeter apparatus which causes a piece
of standard of white cloth (starch free 96×100 cotton fabrics long type) to rub against
the colored specimen under controlled condition of pressure and speed. The rubbing
fingers are covered with white cloth, both for the dry test and wet test, and slight back
and forth for 20 rubbing strokes. The color transferred to the white cloth is compared
with Grey scale. Grey scale for alteration of color, consisting of grade defines the
following:
Observation Grade Qualitative assessment
No dye stain on undyed fabrics 5 Excellent
Slight dye stain on undyed fabrics 4 Very good
Moderate dye stain on undyed fabrics 3 Good
Distinct dye stain on undyed fabrics 2 Fair
Very much dye stain on undyed fabrics 1 Poor
Section-IV: Application
Section-IV: Application 139
The results of light, wash and rubbing fastness properties on silk fabric are tabulated
in Table 31.
Application of Reactive Dyes to Wool
Wool can be dyed with various classes of dyes like acid dyes, metal complex
dyes and reactive dyes. A number of reactive dyes for wool were known before the
discovery of the modern reactive dyes for cellulose[9]
. The first reactive dyes for wool,
Supramine Orange R, were produced in the middle nineteen thirties. It reacts with the
amino group of wool under acidic condition, but it was not realized at the time that it
was chemically combined, although it had high wash fastness. The reactive dyes
contain sulphonic acid groups and therefore they behave as anionic dyes under acidic
conditions. When wool is kept in contact with a solution of a mineral acid for
sufficiently long time equilibrium is attained and positive sites (-+NH2-) are created.
The higher acid concentration is the larger is the number of such sites produced in the
fibre. There is general belief that both the sites (-+NH2-) on the fibre and C=O group
of the amide repeat unit of polyamide fibre are responsible for holding the dye
molecule at various sites on the polymer chain. The interactions supposed to operate
in such attachment are presented below.
Section-IV: Application
Section-IV: Application 140
It is logical imagine that all –NH– of polyamide will not be protonated and all
protonated sites would not be associated with a dye. Sulphonate ion association of dye
anion with –NH– will be stronger than that of SO42Ө
and ClӨ
ion. However, addition
of salt would decrease the extent of this association because of its high concentration.
Hence in cases where it is necessary to level the dyeing, salt is also added to the dye
bath which is acidic. However, the effect of addition of salt will favor the rate of
dyeing at neutral pH though it will adversely affect the dye bath exhaustion at
equilibrium. Increase in the temperature increases the rate of dyeing but as expected it
would decrease the extent of equilibrium dyeing with increase in temperature.
Experimental
This section deals with the application of reactive dyes on wool fabrics. This
section is divided into four subsections dealing respectively with the application of the
dye, study of exhaustion of dye bath, study of the fixation of dye on the wool and the
study of the fastness properties of the dyed patterns.
(2)(i) Application of reactive dyes to wool fabrics
Dyeing with all the reactive dye was carried out by following the same
procedure in all details.
Materials and conditions for 2 % shade
Weight of the wool : 2.0 gm
Amount of the dye under study : 40 mg
Glauber’s salt solution (20 % w/v) : 1.5 ml
Acetic acid solution (10 % w/v) : 1.5 ml
MLR : 1:40
Total volume of the solution in dye bath : 80 ml
pH of the dye bath : 3.0
Dyeing temperature : 100ºC
Time for dyeing : 90 min.
The steps involved are described below:
Section-IV: Application
Section-IV: Application 141
Preparation of wool for dyeing
The procedure described in literature was followed for this purpose[10]
. Wool
(2.0 g) was scoured in a solution of soap (0.2 g) and ammonia (0.05 ml) in water
(100 ml) at 45ºC for 10 minutes. The wool was removed from the bath and rinsed
several times with water. The wool was allowed to stand overnight in distilled water,
squeezed, dried and allowed to condition for two days in laboratory.
Adjustment of pH of wool to a required value
In order to study dyeing at a certain pH, it is necessary to adjust pH of the
suspended wool and solution forming dye bath to a required value. In order to obtain
evenness of pH throughout the material before it enters the dye liquor, a weighted
amount of previously treated wool was heated for 5 min. at 95-100ºC in a solution
which was adjusted to pH 3.0 by adding formic acid solution (10 % w/v). This
treatment was continued till pH of the solution remained fixed at pH 3.0. The wool
were removed and squeezed as far as possible to make free it from adhering mother
liquor.
Dyeing procedure
The dyestuff under study was dissolved by pasting up in cold water and then
addition of hot water. Anhydrous Glauber’s salt solution (1.5 ml, 20% w/v) was added
to it. The pH of the dye bath was adjusted to 3.0 by adding acetic acid solution
(1.5 ml, 10% v/v) and the total volume was adjusted to 80 ml by adding required
amount of distilled water. The temperature of the dye bath was raised to 50ºC and the
wool pattern, which was previously adjusted to pH 3.0, was introduced into the dye
liquor with stirring.
The temperature of the dye bath was gradually increased to 100ºC over a
period of 30 min. and this temperature was maintained for another 30 min.
After this, the dye liquor was decanted into a 250 ml volumetric flask. The
dyed pattern was then washed several times with cold water (100 ml). The washings
were transferred to the volumetric flask. The combined solution of the dye liquor and
washing was then diluted to 250 ml with water. 5 ml of this solution was further
diluted to 25 ml with water and the absorbance of this solution was measured.
The above dyed-pattern was further rinsed with water and dried and a part of it
was mounted on the shade cards 3 and 4.
Section-IV: Application
Section-IV: Application 142
(ii) Exhaustion study of the dye bath
The percentage dye bath exhaustion was calculated following the procedure
described earlier for the silk fabric. The results of percentage dye bath exhaustion are
shown in Table-29. The relevant calibration curves for different dyes are shown in
Figure-37, 38, 39 and 40 and the data are presented in Table-26.
(iii) Fixation study of dyed wool
The percentage fixation of the dyed yarn was calculated following the
procedure described earlier for silk fabric. The results of percentage fixation are
shown in Table-29. The relevant calibration curves for different dyes are shown in
Figure-41, 42, 43 and 44 and the data are presented in Table-27.
(iv) Study of light, wash and rubbing fastness properties of dyed wool
The light, wash and rubbing fastness properties of all the dyed patterns were
studied following the procedure described earlier for silk fabric. The results of light,
wash and rubbing fastness properties on the wool are tabulated in Table-32.
Application of Reactive Dyes to Cotton Fabrics
The attempts to improve wet fastness of dyes on cotton fabric were mainly in
their direction of converting soluble substance into less soluble substances. This led to
dyes, which could be applied to cotton with difficulty. Direct dyes lack wash fastness
because the forces, which retain them on fibre, are easily broken. This suggested that
only good and efficient way to improve wash fastness is to fasten the dye molecule to
cellulose by a covalent bond. Such dyes are designated as reactive dyes
Experimental
This section deals with the application of the reactive dyes on cotton. This
section is divided into four subsections dealing respectively with the application of the
dye, study of exhaustion of dye-bath, study of fixation of dye on the fabrics and the
study of the fastness properties of the dyed patterns.
(i) Application of Reactive Dyes to Cotton fabrics
Dyeing with all the reactive dyes to cotton fabric were carried out by the
following procedure:
Section-IV: Application
Section-IV: Application 143
Materials and conditions for 2% shade
Weight of the cotton fabric : 2.0 gm
Amount of the dye under study : 40 mg
Glauber’s salt solution (20 % w/v) : 1.0 ml
Soda ash solution (10 % w/v) : 1.0 ml
Sodium chloride : 0.5 g
MLR : 1:40
Total volume of the solution in dye bath : 80 ml
pH of the dye bath : 8.0
Dyeing temperature : 100oC
Time for dyeing : 90 min.
This involves two steps:
(a) Pretreatment of the fabric
(b) Dyeing
(a) Pretreatment of the fabric
Cotton fabric (2.0 g) was scoured in a solution of soap (0.2 g) and ammonia
(0.05 ml) in water (100 ml) at 50ºC for 10 min. The fabric was removed from the bath
and rinsed several times with water, squeezed, dried and conditioned for two days in
atmospheric condition and subsequently used for dyeing.
(b)Dyeing procedure
The dyestuff under study was dissolved by pasting up in cold water and then
addition of hot water. Anhydrous Glauber’s salt solution (1.0 ml, 20% w/v) and
sodium chloride (0.5 g) was added to it. The pH of the dye bath was adjusted to 8.0 by
adding soda ash solution (1.0 ml, 4% w/v) and the total volume was adjusted to 80 ml
by adding required amount of distilled water. The temperature of the dye bath was
raised to 40ºC and the cotton pattern was introduced into the dye liquor with stirring.
The temperature of the dye bath was gradually increased to 100ºC over a period of 30
min. and this temperature was maintained for another 60 min.
After this, the dye liquor was decanted into a 250 ml volumetric flask. The
dyed pattern was then washed several times with cold water (100 ml). The washing
was transferred to the volumetric flask. The combined solution of the dye liquor and
Section-IV: Application
Section-IV: Application 144
washing was then diluted to 250 ml with water. 5 ml of this solution was further
diluted to 25 ml with water and the absorbance of this solution was measured.
The above dyed-pattern was further rinsed with water and dried and a part of it
was mounted on the shade cards 5 and 6.
(ii) Exhaustion study of dye bath
The percentage dye bath exhaustion was calculated following the procedure
described earlier for the silk fabric. The results of percentage dye bath exhaustion are
shown in Table-30. The relevant calibration curves for different dyes are shown in
Figure- 37, 38, 39 and 40 and the data are presented in Table-26.
(iii) Fixation study of the dyed cotton fabric
The percentage fixation of the dyed fabric was calculated following the
procedure described earlier for silk fabric. The results of percentage fixation are
shown in Table-30. The relevant calibration curves for different dyes are shown in
Figure-41, 42, 43 and 44 and the data are presented in Table-27.
(iv) Study of light, wash and rubbing fastness properties of dyed cotton fabric
The light, wash and rubbing fastness properties of all the dyed patterns were
studied following the procedure described earlier for silk fabric. The results of light,
wash and rubbing fastness properties on the cotton fabric are tabulated in Table-33.
Section-IV: Application
Section-IV: Application 145
Figure-37: Calibration curve for exhaustion study of reactive dyes for Series-1
0 2 4 6 8 10 12 14 16 18
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45A
bso
rban
ce
Concentration x 10-3(mg/ml)
D1
D3
D5
D7
D9
Figure-38: Calibration curve for exhaustion study of reactive dyes for Series–2
0 2 4 6 8 10 12 14 16 18
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
Abso
rban
ce
Concentration x 10-3 (mg/ml)
D11
D13
D15
D17
D19
Section-IV: Application
Section-IV: Application 146
Figure-39: Calibration curve for exhaustion study of reactive dyes for Series-3
0 2 4 6 8 10 12 14 16 18
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45A
bso
rban
ce
Concentration x 10-3
(mg/ml)
D21
D23
D25
D27
D29
Figure-40: Calibration curve for exhaustion study of reactive dyes for Series-4
0 2 4 6 8 10 12 14 16 18
0.0
0.1
0.2
0.3
0.4
0.5
Abso
rban
ce
Concentration x 10-3 (mg/ml)
D31
D33
D35
D37
D39
Section-IV: Application
Section-IV: Application 147
Table-26: Calibration data for exhaustion study of reactive dyes
Substrate for dyeing : Silk (2.0 g), Wool (2.0 g) & Cotton (2.0 g)
Medium of Spectral study : Aqueous
Dye
No.
Wavelength
(nm)
Absorbance of dye solution at
specified wavelength
Slope of
linear
plot
K
Concentration ×10-3
(mg/ml)
4.0 8.0 12.0 16.0
D1 518 0.085 0.175 0.265 0.350 22.0
D3 490 0.100 0.205 0.280 0.410 25.0
D5 452 0.062 0.125 0.195 0.260 16.7
D7 438 0.070 0.145 0.218 0.289 17.9
D9 465 0.075 0.158 0.240 0.315 20.0
D11 535 0.052 0.108 0.160 0.215 13.3
D13 478 0.075 0.152 0.228 0.302 18.8
D15 455 0.068 0.138 0.205 0.272 17.2
D17 435 0.090 0.178 0.266 0.355 21.9
D19 442 0.096 0.192 0.288 0.385 24.0
D21 525 0.082 0.168 0.252 0.340 22.0
D23 473 0.095 0.190 0.286 0.380 23.1
D25 430 0.102 0.210 0.312 0.418 26.0
D27 442 0.078 0.152 0.232 0.310 20.0
D29 462 0.070 0.142 0.212 0.288 17.9
D31 523 0.110 0.220 0.328 0.442 26.0
D33 494 0.075 0.152 0.230 0.308 18.8
D35 425 0.095 0.192 0.285 0.382 22.7
D37 442 0.065 0.132 0.200 0.265 16.6
D39 275 0.082 0.162 0.242 0.322 20.0
Absorbance = Constant (K) × Concentration
Section-IV: Application
Section-IV: Application 148
Figure-41: Calibration curve for fixation study of reactive dyes for Series-1
0 2 4 6 8 10 12 14 16 18
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40A
bso
rban
ce
Concentration x 10-3
(mg/ml)
D1
D3
D5
D7
D9
Figure-42: Calibration curve for fixation study of reactive dyes for Series-2
0 2 4 6 8 10 12 14 16 18
0.0
0.1
0.2
0.3
0.4
0.5
Abso
rban
ce
Concentration x 10-3 (mg/ml)
D11
D13
D15
D17
D19
Section-IV: Application
Section-IV: Application 149
Figure-43: Calibration curve for fixation study of reactive dyes for Series-3
0 2 4 6 8 10 12 14 16 18
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40A
bso
rban
ce
Concentration x 10-3
(mg/ml)
D21
D23
D25
D27
D29
Figure-44: Calibration curve for fixation study of reactive dyes for Series-4
0 2 4 6 8 10 12 14 16 18
0.0
0.1
0.2
0.3
0.4
0.5
Abso
rban
ce
Concentration x 10-3 (mg/ml)
D31
D33
D35
D37
D39
Section-IV: Application
Section-IV: Application 150
Table-27: Calibration data for fixation study of reactive dyes
Substrate for dyeing : Silk (2.0 g), Wool (2.0 g) & Cotton (2.0 g)
Medium of Spectral study : Conc. Sulphuric acid
Dye
No.
Wavelength
(nm)
Absorbance of dye solution at
specified wavelength
Slope of
linear
plot
K
Concentration ×10-3
(mg/ml)
4.0 8.0 12.0 16.0
D1 515 0.080 0.165 0.245 0.330 20.8
D3 488 0.095 0.190 0.285 0.378 23.7
D5 462 0.088 0.175 0.264 0.352 21.9
D7 435 0.070 0.142 0.212 0.285 17.9
D9 475 0.064 0.130 0.195 0.258 16.7
D11 538 0.100 0.171 0.292 0.392 25.0
D13 470 0.075 0.152 0.232 0.313 19.4
D15 465 0.110 0.220 0.328 0.438 26.0
D17 430 0.085 0.170 0.255 0.342 21.4
D19 448 0.070 0.145 0.218 0.290 18.8
D21 520 0.090 0.180 0.270 0.360 22.7
D23 478 0.098 0.195 0.292 0.390 25.0
D25 434 0.075 0.149 0.220 0.292 17.0
D27 448 0.080 0.158 0.238 0.292 19.5
D29 460 0.068 0.135 0.200 0.268 16.6
D31 528 0.100 0.195 0.290 0.390 23.3
D33 490 0.115 0.212 0.320 0.428 26.0
D35 432 0.080 0.165 0.250 0.334 21.7
D37 445 0.058 0.118 0.175 0.235 15.0
D39 278 0.068 0.138 0.201 0.270 17.9
Absorbance = Constant (K) × Concentration
Section-IV: Application
Section-IV: Application 151
Table- 28: Results of exhaustion and fixation study of reactive dyes
Substrate for dyeing : Silk (2.0 g)
Dyed-pattern for fixation study : Silk (0.1 g)
Amount of dye under study : 40 mg
Medium of spectral study : Water in exhaustion study &
Conc. Sulphuric acid in fixation study
Dye
No.
Amount of
dye
remained
in dye
bath
X (mg)
Amount of
dye
exhausted
in dye bath
40 – X = Y
(mg)
%Ea=
Y x100
40
(mg)
Amount of
dye
in 0.1 g of
dyeing
a
(mg)
Amount of
dye
in 2.0 g of
dyeing
Total wt.
20 a = Z
(mg)
%Fb=
Z x 100
Y
(mg)
D1 3.89 36.11 90.28 1.42 28.4 78.65
D2 4.47 35.53 88.83 1.36 27.2 76.56
D3 4.83 35.17 87.93 1.33 26.6 75.63
D4 5.55 34.45 86.13 1.30 26.0 75.47
D5 4.90 35.10 87.75 1.31 26.2 74.64
D6 5.70 34.30 85.75 1.30 26.0 75.80
D7 6.04 33.96 84.90 1.31 26.2 77.15
D8 5.73 34.27 85.68 1.31 26.2 76.45
D9 5.31 34.69 86.73 1.31 26.2 75.52
D10 4.49 35.51 88.78 1.33 26.6 74.91
D11 4.34 35.66 89.15 1.43 28.6 80.20
D12 3.95 36.05 90.13 1.42 28.4 78.78
D13 3.76 36.24 90.60 1.43 28.6 78.92
D14 4.74 35.26 88.15 1.34 26.8 76.00
D15 4.95 35.05 87.63 1.33 26.6 75.89
D16 5.30 34.70 86.75 1.32 26.4 76.10
D17 5.96 34.04 85.10 1.30 26.0 76.38
D18 5.57 34.43 86.08 1.29 25.8 74.93
D19 5.86 34.14 85.35 1.28 25.6 74.98
D20 6.03 33.97 84.92 1.26 25.2 74.18
Section-IV: Application
Section-IV: Application 152
Continue…
Dye
No.
Amount of
dye
remained
in dye
bath
X (mg)
Amount of
dye
exhausted
in dye bath
40 – X = Y
(mg)
%Ea=
Y x100
40
(mg)
Amount of
dye
in 0.1 g of
dyeing
a
(mg)
Amount of
dye
in 2.0 g of
dyeing
Total wt.
20 a = Z
(mg)
%Fb=
Z x 100
Y
(mg)
D21 3.92 36.08 90.20 1.45 28.9 80.38
D22 4.14 35.86 89.65 1.43 28.6 79.75
D23 4.49 35.51 88.78 1.40 28.0 78.85
D24 5.45 34.55 86.38 1.34 26.8 77.57
D25 5.64 34.36 85.90 1.35 27.0 78.58
D26 5.30 34.70 86.75 1.33 26.6 76.66
D27 4.55 35.45 88.63 1.37 27.4 77.29
D28 5.52 34.48 86.20 1.33 26.6 77.15
D29 4.94 35.06 87.65 1.34 26.8 76.44
D30 5.68 34.32 85.80 1.30 26.0 75.76
D31 3.71 36.29 90.73 1.46 29.2 80.46
D32 3.98 36.02 90.06 1.44 28.8 79.96
D33 4.14 35.86 89.66 1.42 28.4 79.20
D34 4.66 35.34 88.35 1.38 27.6 78.10
D35 5.74 34.26 85.65 1.33 26.6 77.64
D36 5.55 34.45 86.13 1.30 26.0 75.47
D37 5.94 34.06 85.15 1.30 26.0 76.34
D38 5.32 34.68 86.70 1.33 26.6 76.70
D39 4.54 35.46 88.65 1.34 26.8 75.58
D40 5.67 34.33 85.83 1.31 26.2 76.32
Abbreviations: a-Exhaustion, b-Fixation
Section-IV: Application
Section-IV: Application 153
Table-29: Results of Exhaustion and Fixation study of Reactive dyes
Substrate for dyeing : Wool (2.0 g)
Dyed-pattern for fixation study : Wool (0.1 g)
Amount of dye under study : 40 mg
Medium of spectral study : Water in exhaustion study &
Conc. Sulphuric acid in fixation study
Dye
No.
Amount of
dye
remained
in dye
bath
X (mg)
Amount of
dye
exhausted
in dye bath
40 – X = Y
(mg)
%Ea=
Y x100
40
(mg)
Amount of
dye
in 0.1 g of
dyeing
a
(mg)
Amount
of dye
in 2.0 g of
dyeing
Total wt.
20 a = Z
(mg)
%Fb=
Z x 100
Y
(mg)
D1 3.49 36.51 91.28 1.48 29.6 81.07
D2 4.56 35.44 88.60 1.43 28.6 80.70
D3 5.10 34.90 87.25 1.39 27.8 79.66
D4 5.54 34.46 86.15 1.35 27.0 78.35
D5 5.15 34.85 87.13 1.33 26.6 76.33
D6 5.28 34.72 86.80 1.32 26.4 76.04
D7 4.58 35.42 88.55 1.36 27.2 76.79
D8 5.12 34.88 87.20 1.36 27.2 77.98
D9 6.22 33.78 84.45 1.34 26.8 79.34
D10 5.50 34.50 86.25 1.33 26.6 77.10
D11 4.17 35.83 89.58 1.48 29.6 82.61
D12 4.78 35.22 88.06 1.41 28.2 80.07
D13 5.79 34.21 85.53 1.33 26.6 77.76
D14 5.11 34.89 87.23 1.34 26.8 76.81
D15 4.98 35.02 87.55 1.32 26.4 75.39
D16 4.75 35.25 88.13 1.37 27.4 77.73
D17 5.78 34.22 85.55 1.32 26.4 77.15
D18 6.35 33.65 84.13 1.28 25.6 76.08
D19 5.52 34.48 86.20 1.36 27.2 78.88
D20 5.16 34.84 87.10 1.38 27.6 79.22
Section-IV: Application
Section-IV: Application 154
Continue….
Dye
No.
Amount of
dye
remained
in dye bath
X (mg)
Amount of
dye
exhausted
in dye bath
40 – X = Y
(mg)
%Ea=
Y x100
40
(mg)
Amount of
dye
in 0.1 g of
dyeing
a
(mg)
Amount
of dye
in 2.0 g of
dyeing
Total wt.
20 a = Z
(mg)
%Fb=
Z x 100
Y
(mg)
D21 3.80 36.20 90.50 1.49 29.8 82.32
D22 4.19 35.81 89.53 1.46 29.2 81.54
D23 4.49 35.51 88.78 1.43 28.6 80.54
D24 5.66 34.34 85.85 1.37 27.4 79.79
D25 5.46 34.54 86.35 1.36 27.2 78.75
D26 4.98 35.02 87.55 1.35 27.0 77.10
D27 5.50 34.50 86.26 1.31 26.2 75.94
D28 4.59 35.41 88.53 1.39 27.8 78.51
D29 4.96 35.04 87.60 1.32 26.4 75.34
D30 5.40 34.60 86.50 1.36 27.2 78.61
D31 3.89 36.11 90.28 1.47 29.4 81.42
D32 4.00 36.00 90.00 1.45 29.0 80.56
D33 4.55 35.45 88.63 1.41 28.2 79.55
D34 4.93 35.07 87.68 1.38 27.6 78.70
D35 5.42 34.58 86.46 1.30 26.0 75.19
D36 4.70 35.30 88.25 1.35 27.0 76.49
D37 5.00 35.00 87.50 1.36 27.2 77.71
D38 4.61 35.39 88.48 1.35 27.0 76.29
D39 5.92 34.08 85.20 1.29 25.8 75.70
D40 5.56 34.44 86.10 1.35 27.0 78.40
Abbreviations: a-Exhaustion, b-Fixation
Section-IV: Application
Section-IV: Application 155
Table- 30: Results of exhaustion and fixation study of reactive dyes
Substrate for dyeing : Cotton (2.0 g)
Dyed-pattern for fixation study : Cotton (0.1 g)
Amount of dye under study : 40 mg
Medium of spectral study : Water in exhaustion study &
Conc. Sulphuric acid in fixation study
Dye
No.
Amount of
dye
remained
in dye
bath
X (mg)
Amount of
dye
exhausted
in dye bath
40 – X = Y
(mg)
%Ea=
Y x100
40
(mg)
Amount of
dye
in 0.1 g of
dyeing
a
(mg)
Amount
of dye
in 2.0 g of
dyeing
Total wt.
20 a = Z
(mg)
%Fb=
Z x 100
Y
(mg)
D1 4.15 35.85 89.62 1.45 29.0 80.90
D2 5.20 34.80 87.00 1.38 27.6 79.31
D3 6.15 33.85 84.62 1.32 26.4 78.00
D4 4.65 35.35 88.37 1.29 25.8 72.98
D5 5.94 34.06 85.15 1.28 25.6 75.16
D6 5.15 34.85 87.13 1.29 25.8 74.03
D7 6.94 33.06 82.65 1.28 25.6 77.43
D8 6.72 33.28 83.20 1.27 25.4 76.32
D9 4.75 35.25 88.13 1.39 27.8 78.87
D10 4.33 35.67 89.18 1.32 26.4 74.01
D11 3.90 36.10 90.25 1.44 28.8 79.78
D12 4.66 35.34 88.35 1.38 27.6 78.01
D13 5.94 34.06 85.15 1.30 26.0 76.34
D14 6.35 33.65 84.13 1.25 25.0 74.29
D15 5.50 34.50 86.25 1.32 26.4 76.52
D16 4.94 35.06 87.65 1.36 27.2 77.58
D17 6.21 33.79 84.48 1.26 25.2 74.58
D18 6.50 33.50 83.76 1.25 25.0 74.63
D19 6.34 33.66 84.16 1.27 25.4 75.46
D20 5.35 34.65 86.63 1.37 27.4 79.08
Section-IV: Application
Section-IV: Application 156
Continue…
Dye
No.
Amount of
dye
remained
in dye
bath
X (mg)
Amount of
dye
exhausted
in dye
bath
40 – X = Y
(mg)
%Ea=
Y x100
40
(mg)
Amount of
dye
in 0.1 g of
dyeing
a
(mg)
Amount
of dye
in 2.0 g of
dyeing
Total wt.
20 a = Z
(mg)
%Fb=
Z x 100
Y
(mg)
D21 5.10 34.90 87.25 1.36 27.20 77.94
D22 5.95 34.05 85.13 1.28 25.60 75.18
D23 5.34 34.66 86.65 1.31 26.20 75.59
D24 6.49 33.51 83.78 1.30 26.00 77.59
D25 5.34 34.66 86.65 1.32 26.40 76.17
D26 4.54 35.46 88.65 1.40 28.00 78.96
D27 4.95 35.05 87.63 1.34 26.80 76.46
D28 5.22 34.78 86.95 1.32 26.40 75.91
D29 5.64 34.36 85.90 1.28 25.60 74.51
D30 6.05 33.95 84.88 1.28 25.60 75.41
D31 4.35 35.65 89.13 1.43 28.60 80.22
D32 4.70 35.30 88.25 1.40 28.00 79.32
D33 5.46 34.54 86.35 1.36 27.20 78.75
D34 6.35 33.65 84.13 1.29 25.80 76.67
D35 5.50 34.50 86.25 1.33 26.60 77.10
D36 5.95 34.05 85.13 1.30 26.00 76.36
D37 6.10 33.90 84.75 1.27 25.40 74.93
D38 5.50 34.50 86.25 1.29 25.80 74.78
D39 7.15 32.85 82.13 1.25 25.00 76.10
D40 4.70 35.30 88.26 1.34 26.80 75.92
Abbreviations: a-Exhaustion, b-Fixation
Section-IV: Application
Section-IV: Application 157
Table-31: Fastness properties of reactive dyes on silk fabrics
Dye
No.
Fastness properties Dye
No.
Fastness properties
Light Wash Rubbing Light Wash Rubbing
Dry Wet Dry Wet
D1 4-5 5 5 4-5 D21 4 5 5 4-5
D2 5 4-5 3-4 4 D22 5 5 4-5 4
D3 4-5 4 4 3-4 D23 4-5 4 4 4
D4 4 3-4 3 3 D24 3 3-4 3 3-4
D5 6 3 3-4 3 D25 6 3 3-4 3
D6 3-4 4-5 4 3-4 D26 4-5 3-4 4 3
D7 6 3-4 3-4 4 D27 6 4 3 3-4
D8 6 3 3 4 D28 6 3 3-4 4
D9 3-4 3 4-5 3-4 D29 4-5 4 4 4
D10 6 4-5 4 4 D30 6 3-4 3-4 3-4
D11 4 5 5 5 D31 3-4 5 4-5 5
D12 5 4-5 4 4-5 D32 5 4-5 4 4-5
D13 4-5 4 4-5 4 D33 4-5 4-5 4 4
D14 4-5 3-4 3 3 D34 3-4 4 3 3
D15 6 3 3-4 3 D35 6 3 3-4 3-4
D16 3-4 3 4 3-4 D36 4 3-4 3 3
D17 6 4 3 3-4 D37 6 3 3 3-4
D18 6 3-4 3-4 4 D38 6 4 3-4 3
D19 4-5 4 4 3-4 D39 4-5 3-4 4 3-4
D20 6 4-5 3 4 D40 6 4 3-4 4
Light fastness: 1-poor, 2-slight, 3-moderate, 4-fair, 5-good, 6-very good.
Wash & Rubbing fastness: 1-poor, 2-fair, 3-good, 4-very good, 5-excellent.
Section-IV: Application
Section-IV: Application 158
Table-32: Fastness properties of reactive dyes on wool fabrics
Dye
No.
Fastness properties Dye
No.
Fastness properties
Light Wash Rubbing Light Wash Rubbing
Dry Wet Dry Wet
D1 4-5 5 4-5 5 D21 4-5 5 5 5
D2 5-6 4 4 4 D22 5 4-5 4 4-5
D3 5 4 3-4 3-4 D23 4-5 4-5 3-4 4
D4 4-5 3-4 4 3 D24 3-4 4 3 3
D5 6 4 3-4 3-4 D25 6 3 4 3-4
D6 3-4 3 3 3 D26 4-5 4 3 3-4
D7 6 4 4 3-4 D27 6 3-4 3-4 3
D8 6 3-4 3-4 3 D28 6 3 3 4
D9 4-5 4 4 3-4 D29 4-5 3-4 3-4 3-4
D10 6 4 4-5 4-5 D30 6 3 4 4
D11 6 5 5 5 D31 6 5 5 5
D12 5-6 5 4-5 4 D32 5-6 4 4-5 4
D13 5 4-5 4 4 D33 5 4 4-5 3-4
D14 4 4 3 3-4 D34 4-5 3 3 3
D15 6 3 3 3-4 D35 6 3-4 4 4
D16 3-4 4 3-4 3 D36 4-5 3 3-4 3
D17 6 4-5 4 3 D37 6 3-4 3-4 3-4
D18 6 3 4 3-4 D38 6 3 3 3-4
D19 4-5 4 3 4 D39 3-4 3-4 4 4
D20 6 5 3-4 4-5 D40 6 3 3-4 4
Light fastness: 1-poor, 2-slight, 3-moderate, 4-fair, 5-good, 6-very good.
Wash & Rubbing fastness: 1-poor, 2-fair, 3-good, 4-very good, 5-excellent.
Section-IV: Application
Section-IV: Application 159
Table-33: Fastness properties of reactive dyes on cotton fabrics:
Dye
No.
Fastness properties Dye
No.
Fastness properties
Light Wash Rubbing Light Wash Rubbing
Dry Wet Dry Wet
D1 5 5 5 5 D21 5 5 5 5
D2 4-5 4-5 4-5 4 D22 4-5 4 4-5 4
D3 5 4 4 4 D23 5 4 4-5 4
D4 4-5 4 4 3-4 D24 3-4 3-4 4 4
D5 6 3-4 3 3 D25 6 3 3-4 3-4
D6 3-4 3 3-4 3-4 D26 3-4 4 3-4 4
D7 6 3 3-4 3 D27 6 3 3-4 3
D8 6 3-4 3 3 D28 6 3-4 3 3
D9 3-4 3-4 4 4 D29 3-4 3-4 3 3-4
D10 6 4 4 4 D30 6 4 3-4 4
D11 6 5 5 5 D31 4-5 5 5 5
D12 5 4-5 5 3-4 D32 4 4-5 4 5
D13 4-5 4 4 3-4 D33 4-5 4 4 4-5
D14 4 4 3-4 3 D34 5 3-4 3-4 3-4
D15 6 3-4 3 3 D35 6 3 4 3-4
D16 3-4 3 3 3-4 D36 3-4 3-4 4 3
D17 6 4 3-4 3 D37 6 3 3 3
D18 6 3-4 3 4 D38 6 3-4 3 3-4
D19 3-4 4 3-4 4 D39 3-4 4 3 3-4
D20 6 3-4 4 4 D40 6 3-4 4-5 3
Light fastness: 1-poor, 2-slight, 3-moderate, 4-fair, 5-good, 6-very good.
Wash & Rubbing fastness: 1-poor, 2-fair, 3-good, 4-very good, 5-excellent.
Section-IV: Application
Section-IV: Application 160
The Computer Color Matching system (CCM)
Colorimetric data (CIELAB data)
The computer color matching (CCM) system consist of a reflectance
spectrophotometer[11]
attached with a computer system. The spectrophotometer
measures % reflectance of sample at an interval of 20 nm in the visible region i.e.
from 400 nm to 700 nm. From % reflectance at different wavelength, a plot of %
reflectance vs. wavelength is obtained. This is called as ‘Reflectance Profile’ of
colored material. The computer color matching system contains software which can
compute number of important material, which includes K/S, CIE color co-ordinates,
color difference between two samples, matching of two samples etc. This parameter
can be briefly defined as follows:
K/S[12]
is based on Kubelka-Munk theory and it is proportional to the
concentration of dye on substrate.
Several equations have been proposed to relate the reflectance of a dyed fabric
to the concentration of the dye in the fibre. Kubelka-Munk equation has been widely
used in color measurement and it relates reflectance (R) to an absorption coefficient
(S). For a uniformly colored opaque sample illuminated with diffuse light.
According to Kubelka-Munk theory, the optical properties of a sample are
described by two values: K is a measure of the light absorption and S is a measure of
the light scattering. On textiles, K is determined primarily by the dyestuff and S only
by the substrate. From the wavelength, Kubelka and Munk calculate the following
relationship for reflectance R of a thick, opaque sample with constant K and S.
K/S = (1– R)2/2R
[13]
In this equation, R is a fraction (e.g., 0.32 for 32% reflectance). The K/S value
at λmax was used as a measure of the color depth.
Since K and S are additives, the coefficient of a mixture can be calculated
from the K and S values of the individual components. Since the fibres too absorb
light, a correction has to be made for the absorption of light by the fibres.
The correction is very small when the total reflectance is more than 50.0% and
is sometimes ignored, when the correction is made.
Section-IV: Application
Section-IV: Application 161
K/S = C × Abs.
Where C is the dye concentration and Abs is the absorption. This linear
relationship holds fairly well at low dye concentrations, but not so at higher
concentrations.
CIE L* a* b* Co-ordinates
[14]
CIE (Commission International Enclarage) is international committee of color.
CIE tristimulus values X, Y, Z define color in numbers. These values precisely
describe the color but do not give idea about how the color would look like. CIE Lab
color Co-ordinates are obtained by non-linear transformation of color order system.
They give idea about how the color will look like.
Thus, Co-ordinates L* represent lightness (ranging from 0 to 100; 0 for black
and 100 for white), a* is the red-green axis [(+) for red, (0) for gray and (–) for green]
and b* is the yellow-blue axis [(+) for yellow, (0) for gray and (-) for blue)].
Expressing undertones:
1. When primary color is red (a +ve), the undertone is either,
yellow (b +ve) or blue (b –ve).
2. Green (a –ve)
then yellow (b +ve) or blue (b –ve).
3. Yellow (b +ve)
Then red (a +ve) or green (a –ve).
4. Blue (b –ve)
Then red (a +ve) or green (a –ve)
5. When the color is white, grey or black undertone is expressed in both a and b.
+ve Red -ve Green
a
+ve Yellow -ve Blue
b
+ve Lighter -ve Darker
L
+ve Brighter -ve Duller
C
Section-IV: Application
Section-IV: Application 162
The data regarding these values are shown in Table-34 for silk fibres,
Table-35 for wool fibres and Table-36 for cotton fibres.
The graph of K/S values all the dyes D1-D40 are shown in Figure-45 to 48 for
silk fibres; Figure-49 to 52 for wool fibres and Figure-53 to 56 for cotton fibres.
The graph of b* vs a* of all the dyes (Series-1 to 4) are shown in Figure-57 to
60 for silk fibre, Figure-61 to 64 for wool fibre and Figures-65 to 68 for cotton fibre.
Color difference equation[15]
:
When two samples do not match, the magnitude of color difference can be
calculated from difference in color co-ordinates. i.e. L* a* b* formula:
E = [(L2) + (a
2) + (b
2)]
1/2
Where E is total color difference. The computer color matching system calculates
color difference using above equation.
Section-IV: Application
Section-IV: Application 163
Table-34: Color difference data and K/S data on Silk fibre.
Dye
No.
L* a* b* C* H* K/S
D1 66.85 23.45 04.96 23.97 11.94 00.969
D2 68.50 26.44 21.58 34.13 39.22 01.451
D3 70.87 22.89 11.26 25.51 26.20 00.765
D4 71.83 21.74 07.54 23.01 19.13 00.711
D5 77.54 15.28 13.28 20.24 41.00 00.478
D6 68.73 25.26 14.82 29.29 30.40 01.066
D7 81.13 12.49 12.18 17.44 44.29 00.468
D8 73.43 17.74 39.02 42.87 65.55 01.913
D9 76.98 16.17 09.66 18.84 30.86 00.471
D10 78.81 14.84 10.62 18.25 35.58 00.431
D11 45.41 54.18 06.36 54.55 06.70 10.758
D12 60.86 39.06 49.14 62.78 51.52 07.650
D13 68.41 29.46 33.98 44.98 49.07 02.523
D14 71.21 22.51 14.56 26.81 32.90 00.907
D15 78.23 13.63 14.94 20.22 47.63 00.474
D16 72.32 22.62 15.34 27.33 34.15 00.836
D17 83.64 05.71 21.36 22.11 75.04 00.431
D18 83.21 05.01 18.38 19.05 74.77 00.411
D19 79.54 14.48 12.34 19.02 40.44 00.401
D20 85.37 02.13 14.02 14.18 81.34 00.334
Section-IV: Application
Section-IV: Application 164
Continue…
Dye
No.
L* a* b* C* H* K/S
D21 60.09 27.96 -06.08 28.61 347.73 01.593
D22 62.03 26.78 20.28 33.59 37.14 02.324
D23 61.67 30.40 13.66 33.32 24.20 02.124
D24 68.82 19.67 05.64 20.46 16.00 00.812
D25 81.16 05.77 14.78 15.87 68.67 00.544
D26 71.11 18.66 09.80 21.08 27.70 01.255
D27 77.38 06.16 26.24 26.95 76.79 01.341
D28 80.40 03.79 20.16 20.51 79.35 00.982
D29 69.54 17.42 06.94 18.75 21.72 00.748
D30 72.93 13.58 25.26 28.68 61.74 01.650
D31 58.75 30.35 -00.78 30.36 358.53 01.890
D32 78.36 14.58 15.72 21.44 47.15 00.504
D33 76.68 16.07 10.68 19.29 33.61 00.488
D34 81.86 13.64 04.82 14.47 19.46 00.253
D35 77.75 10.53 35.06 36.61 73.29 01.226
D36 77.19 16.08 09.02 18.44 29.29 00.441
D37 76.50 10.77 37.72 39.23 74.07 01.520
D38 80.91 06.15 19.64 20.58 72.62 00.605
D39 76.08 15.02 12.56 19.58 39.91 00.530
D40 81.05 12.72 07.84 14.95 31.64 00.284
Section-IV: Application
Section-IV: Application 165
Table-35: Color difference data and K/S data on wool fibre.
Dye
No.
L* a* b* C* H* K/S
D1 42.88 36.05 14.76 38.96 22.26 06.993
D2 58.26 34.31 34.58 48.72 45.22 04.481
D3 52.72 40.07 29.84 49.96 36.68 06.229
D4 33.01 41.71 21.48 46.91 27.25 21.135
D5 57.77 33.32 36.62 49.51 47.70 04.884
D6 51.53 38.34 33.46 50.89 41.11 06.620
D7 61.55 28.66 33.06 43.75 49.08 05.198
D8 57.86 33.07 36.12 48.97 47.53 04.884
D9 36.36 42.71 30.42 52.44 35.46 20.751
D10 55.89 31.44 28.62 42.51 42.32 04.638
D11 32.36 49.00 17.38 51.99 19.53 26.183
D12 53.40 38.02 51.80 64.26 53.72 14.354
D13 51.36 35.65 45.64 57.91 52.01 13.263
D14 43.45 39.93 32.38 51.41 39.04 14.122
D15 55.38 34.92 45.42 57.29 52.44 08.753
D16 50.73 35.27 29.64 46.07 40.04 06.344
D17 69.92 13.67 27.70 30.89 63.73 02.046
D18 64.20 23.17 44.98 50.60 62.75 04.333
D19 67.97 16.54 30.90 35.05 61.85 02.826
D20 57.70 31.55 24.46 39.92 37.78 03.435
Section-IV: Application
Section-IV: Application 166
Continue…
Dye
No.
L* a* b* C* H* K/S
D21 47.76 31.41 09.88 32.93 17.46 06.796
D22 65.84 14.86 31.88 35.18 65.00 05.168
D23 51.95 31.30 29.10 42.74 42.91 07.461
D24 43.00 37.16 26.24 45.49 35.23 11.305
D25 65.31 15.27 36.54 39.60 67.32 07.135
D26 66.62 16.80 31.42 35.63 61.86 06.239
D27 66.00 21.83 42.60 47.87 62.87 04.613
D28 70.25 08.77 39.22 40.19 77.40 07.800
D29 44.09 43.42 43.16 61.22 44.83 23.282
D30 67.57 09.68 35.76 37.05 74.85 10.411
D31 60.48 26.41 10.96 28.59 22.54 04.446
D32 72.23 14.78 27.52 31.24 61.76 04.582
D33 61.17 25.05 24.86 35.29 44.79 05.474
D34 57.20 33.01 20.66 38.94 32.04 05.044
D35 72.32 15.77 31.04 34.82 63.07 04.182
D36 65.60 23.28 22.68 32.50 44.26 05.016
D37 73.34 12.12 25.62 28.34 64.69 04.172
D38 74.53 10.66 27.96 29.92 69.14 04.208
D39 51.14 25.85 25.12 36.05 44.18 07.405
D40 59.58 33.38 26.34 42.52 38.27 05.369
Section-IV: Application
Section-IV: Application 167
Table-36: Color difference data and K/S data on cotton fibre.
Dye
No.
L* a* b* C* H* K/S
D1 42.93 42.56 10.14 43.75 13.40 08.069
D2 67.59 30.24 24.62 38.99 39.15 01.660
D3 62.71 35.01 22.62 41.68 32.87 02.415
D4 42.06 45.59 20.28 49.90 23.98 11.157
D5 65.92 26.03 27.04 37.53 46.09 01.884
D6 62.71 31.36 32.28 45.01 45.83 03.059
D7 68.68 26.19 30.80 40.43 49.63 02.120
D8 66.30 30.76 26.72 40.75 40.98 01.956
D9 55.81 32.10 26.38 41.55 39.42 03.696
D10 67.08 28.37 21.34 35.50 36.95 01.463
D11 42.05 54.00 11.48 55.21 12.00 13.303
D12 64.17 36.56 50.52 62.36 54.11 06.198
D13 64.30 34.05 38.06 51.07 48.19 03.735
D14 51.39 38.83 31.06 49.72 38.66 07.019
D15 67.14 26.11 25.24 36.31 44.03 01.594
D16 63.97 29.60 24.38 38.35 39.48 01.970
D17 79.22 13.83 27.32 30.62 63.15 00.890
D18 73.65 16.04 35.04 38.54 65.41 01.860
D19 69.89 17.45 29.22 34.03 59.16 02.509
D20 73.23 12.80 20.62 24.27 58.18 01.306
Section-IV: Application
Section-IV: Application 168
Continue…
Dye
No.
L* a* b* C* H* K/S
D21 40.41 37.69 -3.56 37.85 354.60 08.005
D22 72.46 15.43 20.32 25.52 52.79 01.574
D23 60.15 33.03 22.02 39.70 33.69 03.104
D24 54.00 27.70 14.86 31.43 28.21 03.321
D25 75.57 08.60 21.20 22.88 67.22 01.410
D26 69.12 17.00 19.36 25.77 48.71 02.486
D27 73.67 12.81 23.92 27.13 61.83 01.550
D28 65.65 10.50 27.22 29.18 68.91 04.214
D29 47.54 32.81 16.68 36.80 26.95 06.000
D30 73.75 08.92 21.06 22.87 67.04 01.812
D31 52.15 36.59 -00.30 36.60 359.53 03.462
D32 76.31 16.42 21.50 27.05 52.64 01.006
D33 66.14 25.37 18.36 31.32 35.89 01.483
D34 66.60 29.55 13.06 32.30 23.85 01.378
D35 77.41 15.18 25.94 30.06 59.66 01.008
D36 72.48 21.97 15.98 27.17 36.03 01.123
D37 77.65 12.89 25.18 28.29 62.90 01.338
D38 78.98 09.07 21.78 23.59 67.39 00.991
D39 63.80 22.17 17.62 28.32 38.48 01.519
D40 67.78 27.89 18.48 33.46 33.53 01.423
Section-IV: Application
Section-IV: Application 169
Figure-45: Graph of K/S values of dyes D1-10 on silk fibre, which shows that dye D8
shows maximum K/S value and D10 shows minimum K/S value.
D1 D2 D3 D4 D5 D6 D7 D8 D9 D10
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
0.4310.471
1.913
0.468
1.066
0.478
0.7110.765
1.451
0.969
K/S
Dyes (D1-D
10)
Figure-46: Graph of K/S values of dyes D11-20 on silk fibre, which shows that dye D11
shows maximum K/S value and dye D20 shows minimum K/S value.
D11 D12 D13 D14 D15 D16 D17 D18 D19 D20
0
2
4
6
8
10
0.3340.4010.4110.4310.836
0.4740.907
2.523
7.650
10.758
K/S
Dyes (D11
-D20
)
Section-IV: Application
Section-IV: Application 170
Figure-47: Graph of K/S values of dyes D21-30 on silk fibre, which shows that dye D22
shows maximum K/S value and dye D25 shows minimum K/S value.
D21 D22 D23 D24 D25 D26 D27 D28 D29 D30
0.0
0.5
1.0
1.5
2.0
2.5
1.65
0.748
0.982
1.3411.255
0.544
0.812
2.124
2.324
1.593
K/S
Dyes (D21
-D30
)
Figure-48: Graph of K/S values of dyes D31-40 on silk fibre, which shows that dye D31
shows maximum K/S value and dye D34 shows minimum K/S value.
D31 D32 D33 D34 D35 D36 D37 D38 D39 D40
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
0.284
0.5300.605
1.520
0.441
1.226
0.253
0.4880.504
1.890
K/S
Dyes (D31
-D40
)
Section-IV: Application
Section-IV: Application 171
Figure-49: Graph of K/S values of dyes D1-10 on wool fibre, which shows that dye D4
shows maximum K/S value and dye D2 shows minimum K/S value.
D1 D2 D3 D4 D5 D6 D7 D8 D9 D10
0
5
10
15
20
4.638
20.751
4.8845.198
6.620
4.884
21.135
6.229
4.481
6.993
K/S
Dyes (D1-D
10)
Figure-50: Graph of K/S values of dyes D11-20 on wool fibre, which shows that dye
D11 shows maximum K/S value and dye D17 shows minimum K/S value.
D11 D12 D13 D14 D15 D16 D17 D18 D19 D20
0
5
10
15
20
25
3.4352.826
4.333
2.046
6.344
8.753
14.12213.263
14.354
26.183
K/S
Dyes (D11
-D20
)
Section-IV: Application
Section-IV: Application 172
Figure-51: Graph of K/S values of dyes D21-30 on wool fibre, which shows that dye
D29 shows maximum K/S value and dye D27 shows minimum K/S value.
D21 D22 D23 D24 D25 D26 D27 D28 D29 D30
0
5
10
15
20
25
10.411
23.282
7.800
4.613
6.2397.135
11.305
7.461
5.168
6.796
K/S
Dyes (D21
-D30
)
Figure-52: Graph of K/S values of dyes D31-40 on wool fibre, which shows that dye
D39 shows maximum K/S value and dye D37 shows minimum K/S value.
D31 D32 D33 D34 D35 D36 D37 D38 D39 D40
0
1
2
3
4
5
6
7
5.369
7.405
4.2084.172
5.016
4.182
5.044
5.474
4.5824.446
K/S
Dyes (D31
-D40
)
Section-IV: Application
Section-IV: Application 173
Figure-53: Graph of K/S values of dyes D1-10 on cotton fibre, which shows that dye
D4 shows maximum K/S value and dye D10 shows minimum K/S value.
D1 D2 D3 D4 D5 D6 D7 D8 D9 D10
0
2
4
6
8
10
12
1.463
3.696
1.9562.120
3.059
1.884
11.157
2.415
1.660
8.069
K/S
Dyes (D1-D
10)
Figure-54: Graph of K/S values of dyes D11-20 on cotton fibre, which shows that dye
D11 shows maximum K/S value and dye D17 shows minimum K/S value.
D11 D12 D13 D14 D15 D16 D17 D18 D19 D20
0
2
4
6
8
10
12
14
1.306
2.5091.86
0.890
1.9701.594
7.019
3.735
6.198
13.303
K/S
Dyes (D11
-D20
)
Section-IV: Application
Section-IV: Application 174
Figure-55: Graph of K/S values of dyes D21-30 on cotton fibre, which shows that dye
D21 shows maximum K/S value and dye D25 shows minimum K/S value.
D21 D22 D23 D24 D25 D26 D27 D28 D29 D30
0
1
2
3
4
5
6
7
8
1.812
6.000
4.214
1.550
2.486
1.410
3.3213.104
1.574
8.005
K/S
Dyes (D21
-D30
)
Figure-56: Graph of K/S values of dyes D31-40 on cotton fibre, which shows that dye
D31 shows maximum K/S value and dye D38 shows minimum K/S value.
D31 D32 D33 D34 D35 D36 D37 D38 D39 D40
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
1.4231.519
0.991
1.338
1.1231.008
1.3781.483
1.006
3.462
K/S
Dyes (D31
-D40
)
Section-IV: Application
Section-IV: Application 175
Figure-57: Graph of b* vs a* for silk fibre (Series-1)
0
5
10
15
20
25
30
35
40
45
0 5 10 15 20 25 30
a*
b*
D1
D2
D3
D4
D5
D6
D7
D8
D9
D10
Figure-58: Graph of b* vs a* for silk fibre (Series-2)
0
10
20
30
40
50
60
0 10 20 30 40 50 60
a*
b*
D11
D12
D13
D14
D15
D16
D17
D18
D19
D20
Figure-59: Graph of b* vs a* for silk fibre (Series-3)
-10
-5
0
5
10
15
20
25
30
0 5 10 15 20 25 30 35
a*
b*
D21
D22
D23
D24
D25
D26
D27
D28
D29
D30
Section-IV: Application
Section-IV: Application 176
Figure-60: Graph of b* vs a* for silk fibre (Series-4)
-5
0
5
10
15
20
25
30
35
40
0 5 10 15 20 25 30 35
a*
b*
D31
D32
D33
D34
D35
D36
D37
D38
D39
D40
Figure-61: Graph of b* vs a* for wool fibre (Series-1)
0
5
10
15
20
25
30
35
40
0 10 20 30 40 50
a*
b*
D1
D2
D3
D4
D5
D6
D7
D8
D9
D10
Figure-62: Graph of b* vs a* for wool fibre (Series-2)
0
10
20
30
40
50
60
0 10 20 30 40 50 60
a*
b*
D11
D12
D13
D14
D15
D16
D17
D18
D19
D20
Section-IV: Application
Section-IV: Application 177
Figure-63: Graph of b* vs a* for wool fibre (Series-3)
0
5
10
15
20
25
30
35
40
45
50
0 10 20 30 40 50
a*
b*
D21
D22
D23
D24
D25
D26
D27
D28
D29
D30
Figure-64: Graph of b* vs a* for wool fibre (Series-4)
0
5
10
15
20
25
30
35
0 10 20 30 40
a*
b*
D31
D32
D33
D34
D35
D36
D37
D38
D39
D40
Figure-65: Graph of b* vs a* for cotton fibre (Series-1)
0
5
10
15
20
25
30
35
0 10 20 30 40 50
a*
b*
D1
D2
D3
D4
D5
D6
D7
D8
D9
D10
Section-IV: Application
Section-IV: Application 178
Figure-66: Graph of b* vs a* for cotton fibre (Series-2)
0
10
20
30
40
50
60
0 10 20 30 40 50 60
a*
b*
D11
D12
D13
D14
D15
D16
D17
D18
D19
D20
Figure-67: Graph of b* vs a* for cotton fibre (Series-3)
-10
-5
0
5
10
15
20
25
30
0 10 20 30 40
a*
b*
D21
D22
D23
D24
D25
D26
D27
D28
D29
D30
Figure-68: Graph of b* vs a* for cotton fibre (Series-4)
-5
0
5
10
15
20
25
30
0 10 20 30 40
a*
b*
D31
D32
D33
D34
D35
D36
D37
D38
D39
D40
Section-IV: Application
Section-IV: Application 179
Antimicrobial activity (Antibacterial and antifungal)
Antibacterial activity
Introduction
Man is closely influenced by the activities if microorganisms. Microorganisms
are a part of our lives in more ways than most of us understand. They have shaped our
present environment and their activities will greatly influence our future.
Microorganisms should not be considered separate from human beings, but profound
beneficial influence as a part of our life. They are employed in the manufacture of
dairy products, certain foods, min processing of certain medicines and therapeutic
agents, in manufacture of certain chemicals and in numerous other ways.
Despite the established useful functions and potentially valuable activities of
microorganism, these microscopic dorms of life may be best known as agents of food
spoilage and causal agents of human beings viz. Acquired immune deficiency
syndrome[aids], herpes, legionnaires disease, influenza, jaundice, tuberculosis,
typhoid, dermatomycoses, dysentery, malaria etc. Animals [infected with brucellosis,
tularemia etc...] and plants [infected with mildews, rusts, smuts, cankers, leaf spots,
etc.] have also been known to be victims of microbial pathogens. So far as is known,
all primitive and civilized societies have experienced diseases caused by microbes,
frequently with disastrous results. Moreover, microorganisms have played profound
roles in warfare, religion and the migration of populations.
Control of microbial population is necessary to prevent transmission of
disease, infection, decomposition, contamination and spoilage caused by them, man’s
personal comforts and convenience depend to a large extent on the control of
microbial population.
Bacteria
In 1928, a German scientist C.E. Chrenberg first used the term “bacterium” to
denote small microscopic organism with a relatively simple and primitive form of the
cellular organization known as “prokaryotic”.
Danish physician, Gram in peculiarity, bacteria are generally unicellular e.g.
cocci, bacilli, etc filamentous, eg. actinomycetes, some being sheathed having certain
cells specialized for reproduction. The microorganisms are capable of producing
diseases in host are known as ‘pathogenic’. Most of the microorganisms present on
Section-IV: Application
Section-IV: Application 180
the skin and mucous membrane are non pathogenic and are often referred to as
“commensals” or if they live on food residues as in intestine, they may be called
“saprophytes”. Generally, the pathogenic cocci and bacilli are Gram positive and the
pathogenic coco bacilli are Gram negative.
For evaluation of antibacterial activity in our case, we have used S. aureus and
S. pyogenes from Gram positive group of bacteria and E. coli and P. aeruginosa from
Gram negative group of bacteria.
S. aureus
Genus: staphylococcus [microccaceae]
Staphylococci are differentiated from micrococcus, a genus of the same family
by its ability to utilize glucose, mannitol and pyruvate anaerobically. Cells of
staphylococci are usually to be found on the skin or mucous membranes of the animal
body, especially of the nose and mouth where they occur in large numbers even under
normal conditions.
Species: S. aureus
The individual cells of S.aureus are 0.8 to 0.9 µ in diameter. They are ovoid or
spherical, non motile, non capsulated, non sporing stain with ordinary aniline dyes
and Gram positive, typically arranged in groups of irregular clusters like branches of
groups found in pus, singly or in pairs. The optimum temperature for the growth us
37ºC, optimum pH is 7.4 to 7.6. They produce golden yellow pigment, which
develops best at room temperature. They cause pyoregenic of pus forming
[suppurative] conditions, mastitis of women and cows, boils, carbuncles infantile
impetigo, internal abscess and food poisoning.
E. coli
Genus: Escherichia [enterobacteriaceae]
This genus comprises Escherichia and several variants and is of particular
interest to the sanitarian since they occur commonly in the formal intestinal tract of
man and animals. Their presence in foods or in drinking water may indicate faecal
pollution. E. coli is the most distinctively recognized feacal species.
Species: Escherichia coli
E. coli is the most important type in this species, which contains a number of
other types. Escherichia in 1885 discovered in from the faces of the newborn and
Section-IV: Application
Section-IV: Application 181
showed the organisms in the intesting within three days after birth. It is a commensals
of the human intesting and found in the intestinal tract of men and animals and is also
found in the sewage water, land, soil contaminated by feacal matters. The Gram
negative rods are 2 to 4 µ by 0.4 µ in size, commonly seen in coccobacillary form and
rarely in filamentous form. They are facultative anaerobes and grow in all laboratory
media. Colonies are circular, raised, smooth and emit a faecal odour. E. coli are
generally non pathogenic and are incriminated as pathogens because in certain
instances some strains have been found to produce septicemia, inflammations of liver
and gall bladder, appendix, meningitis, pneumonia and other infections and this
species is a recognized pathogen in the veterinary field.
S. pyogenes
Genus: Streptococcus
The term streptococcus was first introduced by Bilroth [1874] and the term
S. pyogenes was used by Rosenbach [1884]. These are spherical or ovoid cells; divide
in one axis and form chains; nonmotile and nonsporing. The growth is absence of
native proteins in the medium; they produce characteristic haemolytic changes in
media containing blood; produce acid only by fermentation of carbohydrates; often
fail to liquefy gelatin; some strains produce exotoxin and extracellular products; a few
of them are anaerobic.
Species: Streptococcus pyogenes
S. pyogenes is pathogenic to human and found in sore throat, follicular
tonsillitis, septicemia, acute or malignant ulcerative endocarditis etc. These are
spherical cocci 0.5 to 0.75 micro in diameter, arranged in moderately long chains of
round cocci and easily differentiated from enterococci that from short chains of 2 to 4
spheres. S. pyogenes is recently isolated from throat or other lesions; they show either
mucoid or matt colonies. On keeping in the laboratory, they undergo varation to a
glossy type. Streptococci are susceptible to destructive agents, and to penicillin and
sulphonamides.
Section-IV: Application
Section-IV: Application 182
P. aeruginosa
Genus: Pseudomonas
Genus pseudomonas is characterized by gram negative motile rods,
nonsporing aerobes, oxidase positive, bluish green or yellowish pigment diffusing into
the medium. Out of 140 species, only one is pathogenic to human.
Species: Pseudomonas aeruginosa
P. aeruginosa occurs as a commensal in the intestine of human and animal’s
but, when the defensive mechanism of the body is poor. It acts as a minor pathogen
producing suppurative wound, otitis media, peritonitis, cystitis, bronchopneumonia
and empyema. In children it causes diarrhea and septicemia. The pus produced by
P.aeruginosa is greenish blue. These are gram negative, actively motile, non sporing
organisms 1.5-3.0 micro by 0.5 micro with rounded ends and bipolar flagella. They
occur singly or in pair, of short chains. They grow well in ordinary media under
aerobic conditions, producing diffusible pigment.
Antifungal activity
Introduction
It has been estimated that the life expectancy of humans has increased by at
least 10 years since the discovery of antimicrobial agents for the treatment of
microbial infections. A consequence of our success with antimicrobial agents and
improved medical care is the number of fungal infections.
The incidence of fungal infections has increased dramatically in the past 20
years partly because of the increase in the number of people whose immune systems
are compromised by wither aids, aging, organ transplantation or cancer therapy.
Accordingly, the increase in rates of morbidity and mortality because of fungal
infections has been now recognized as a major problem. In response to the increased
incidence of fungal infections, the pharmaceutical industry has developed a number of
newer less toxic antifungal for clinical use. The increased use of antifungal, often for
prolonged periods, has lead to recognition of the phenomenon of acquired antifungal
resistance to one or more of the available antifungal.
Fungi are nonphotosynthetic eukaryotes growing either as colonies of single
cells (yeasts) or as filamentous multicellular aggregate [molds]. Most fungi live as
Section-IV: Application
Section-IV: Application 183
saprophytes in soil or on dead plant material and are important in the mineralization
of organic matter. A smaller number produce disease in human and animals. The in
vitro methods used for detections of antifungal potency are similar to those used in
antibacterial screening. As with bacteria, it is easy to discover several synthetic and
natural compounds that, in small quantity, can retard or prevent growth of fungi in
culture media.
C. albicans
Genus: Candida
Candida species reproduce by yeast like budding cells but they also show
formation of pseudomycellum. These pseudomycellum are chains of elongated cells
formed from buds and the buds elongated without breaking of the mother cell. They
are very fragile and separate easily. Mycelia also form by the elongation of the germ
tube produced by a mother cell.
Species: Candida albicans
C. albicans may remain as a commensal of the mucous membrane with or
without causing any pathologic changes to the deeper tissues of the same fungus may
cause pathological lesion of the skin. Such a fungus under favorable conditions can
cause superficial, intermediate of deep mycoses depending on the condition of the
host.
A. niger and A. clavatus
Genus: Aspergillus
The aspergilli are widespread in nature, being found on fruits, vegetables and
other substrates, which may provide nutriment. Some species are involved in food
spoilage. They are important economically because they are used in a number of
industrial fermentations, including the production of citric acid gluconic acid.
Aspergilli grow in high concentrations of sugar and salt, indicating that they can
extract water required for their growth from relatively dry substances.
Evaluation techniques
The following conditions must be met for the screening of antimicrobial
activity. There should be intimate contact between the test organisms and substance to
be evaluated.
Section-IV: Application
Section-IV: Application 184
Required conditions should be provided for the growth of microorganisms.
Conditions should be same through the study.
Aseptic/sterile environment should be maintained.
Various methods have been used from time to time by several workers to
evaluate the antimicrobial activity. The evaluation can be done by the following
methods:
1. Turbidometric method
2. Agar streak dilution method
3. Serial dilution method
4. Agar diffusion method
Following techniques are used as agar diffusion method:
1. Agar cup method.
2. Agar ditch method.
3. Paper disc method.
We have used the broth dilution method to evaluate the antibacterial
activity[16]
.
It is one of the non automated in vitro bacterial susceptibility tests.
This classic method yields a quantitative result for the amount of antimicrobial agents
that is needed to inhibit growth of specific microorganisms. It is carried out in tubes.
1. Macro dillution method in tubes
2. Micro dillution format using plastic trays
Determination of minimal bactericidal concentrations by agar cup method
Materials and method
1. All the synthesized drugs were used for antibacterial test procedures
2. All necessary controls like:
2.1 Drug control
2.2 Vehicle control
2.3 Agar control
2.4 Organism control
2.5 Known antibacterial drugs control
2.6 All MTCC cultures were tested against above mentioned known and unknown
drugs.
Section-IV: Application
Section-IV: Application 185
2.7 Mueller hinton broth was used as nutrient medium to grow and dilute the drug
suspension for the test.
2.8 Inoculum size for test strain was adjusted to 108 cfu [colony forming unit] per
milliliter by comparing the turbidity.
Following common standard strains were used for screening of antibacterial
and antifungal activities: the strains were procured from institute of microbial
technology, chandigarh.
E. coli (Gram negative) MTCC – 442
P. aeruginosa (Gram negative) MTCC – 441
S. aureus (Gram positive) MTCC – 96
S. pyogenes (Gram positive) MTCC – 443
C. albicans (fungus) MTCC – 227
A. niger (fungus) MTCC – 282
A. clavatus (fungus) MTCC – 1323
DMSO was used as diluents/vehicle to get desired concentration of drugs to
test upon standard bacterial strains.
Minimal bactericidal concentration [MBC]
The main advantage of the ‘broth dilution method’ for MBC determination
lies in the fact that it can readily be converted to determine the MBC as well.
1. Serial dilutions were prepared in primary and secondary screening.
2. The control tube containing no antibiotic is immediately sub cultured [before
inoculation] by spreading a loopful evenly over a quarter of p[late of medium
suitable for the growth of the test organism and put for incubation at 37ºC
overnight. The tubes are then incubated overnight.
3. The MBC of the control organism is read to check the accuracy of the drug
concentrations.
4. The lowest concentration inhibiting growth of the organism is recorded as the
MBC.
5. All the tubes not showing visible growth [in the same manner as control tube
described above] are sub cultured and incubated overnight at 37ºC.
Section-IV: Application
Section-IV: Application 186
6. The amount of growth from the control tube before incubation [which
represents the original inoculum] is compared.
7. Subcultures may show: similar number of colonies indicating bacteriostatis, a
reduced number of colonies indicating a partial or slow bactericidal activity &
no growth, if the whole inoculum has been killed. The test must include a
second set of the same dilutions inoculated with an organism of known
sensitivity.
Methods used for primary and secondary screening
Each synthesized drug was diluted obtaining 2000 µg/ml concentration, as a
stock solution.
Primary screen
In primary screening 500 µ/ml, 250 µ/ml, and 125 µ/ml concentrations of the
synthesized drugs were taken. The active synthesized drugs found in this primary
screening were further tested in a second set of dilution against all microorganisms.
Secondary screen
The drugs found active in primary screening were similarly diluted to obtain
100 µ/ml, 50 µ/ml, 25 µ/ml, 12.5 µ/ml, 6.250 µ/ml, 3.125 µ/ml and 1.5625 µ/ml
concentrations.
Reading result
The highest dilution showing at least 99% inhibition zone is taken as MIC.
The result of this is much affected by the size of the inoculum. The test mixture
should contain 108 organism/ml.
Reading results
The highest dilution showing at least 99% inhibition is taken as MBC.
The result of this test is much affected by the size of the inoculum. The test
mixture should contain 108 organisms/ ml.
The standard drugs
The standard drug used in the present study is Gentamycin for evaluating
antibacterial activity which showed [0.25, 0.05, 0.5 & 1 µg/ml]. MBC against
Section-IV: Application
Section-IV: Application 187
S. aureus, E. coli, S. pyogenes. & P. aeruginosa respectively. Nystatin is used as the
standard drug for antifungal activity which showed 100 µg/ml. MFC against all the
species, used for the antifungal activity.
Antibacterial activity data and antifungal data of all the reactive dyes (D1-D40)
are summarized in Table-37 and 38.
Section-IV: Application
Section-IV: Application 188
Table-37: Antibacterial activity of reactive dyes D1-D40
Dye No. Minimal Bactericidal Concentration (µg/ml)
Gram-negative Gram-positive
E. coli P. Aeruginosa S. aureus S. pyogenus
MTCC443 MTCC 1688 MTCC 96 MTCC 442
D1 200 500 250 200
D2 500 500 250 500
D3 250 200 500 1000
D4 200 500 500 1000
D5 100 100 100 100
D6 500 500 200 200
D7 100 250 500 500
D8 200 500 250 500
D9 250 200 200 200
D10 100 200 250 250
D11 62.5 200 200 200
D12 500 500 500 1000
D13 100 200 200 200
D14 500 1000 500 500
D15 250 250 500 500
D16 62.5 200 250 500
D17 500 200 500 1000
D18 1000 1000 1000 1000
D19 1000 1000 1000 500
D20 500 500 100 1000
Gentamycin 0.05 1 0.25 0.5
Ampicillin 100 100 250 100
Chloramphenicol 50 50 50 50
Ciprofloxacin 25 25 50 50
Norfloxacin 10 10 10 10
Section-IV: Application
Section-IV: Application 189
Continue….
Dye No. Minimal Bactericidal Concentration (µg/ml)
Gram-negative Gram-positive
E. coli P. Aeruginosa S. aureus S. pyogenus
MTCC443 MTCC 1688 MTCC 96 MTCC 442
D21 500 500 100 1000
D22 100 500 500 500
D23 200 500 500 200
D24 200 200 1000 100
D25 50 100 250 250
D26 200 500 1000 200
D27 200 500 1000 1000
D28 500 500 1000 1000
D29 62.5 100 200 200
D30 200 100 500 500
D31 250 250 500 250
D32 250 500 500 500
D33 500 200 100 100
D34 100 500 100 500
D35 500 100 200 500
D36 100 200 500 500
D37 200 200 1000 1000
D38 500 500 500 500
D39 62.5 100 200 100
D40 100 100 500 100
Gentamycin 0.05 1 0.25 0.5
Ampicillin 100 100 250 100
Chloramphenicol 50 50 50 50
Ciprofloxacin 25 25 50 50
Norfloxacin 10 10 10 10
Section-IV: Application
Section-IV: Application 190
Table-38: Antifungal activity of reactive dyes D1-D40
Dye No. Minimal fungicidal Concentration (µg/ml)
C. albicans A. niger A. clavatus
MTCC 227 MTCC 282 MTCC 1323
D1 200 500 500
D2 500 1000 1000
D3 100 250 250
D4 200 500 500
D5 200 500 500
D6 200 500 500
D7 100 500 500
D8 500 1000 1000
D9 500 >1000 >1000
D10 200 1000 1000
D11 500 >1000 >1000
D12 500 500 500
D13 1000 1000 >1000
D14 100 250 250
D15 1000 1000 >1000
D16 100 250 250
D17 1000 1000 1000
D18 100 250 250
D19 500 >1000 >1000
D20 1000 500 500
Nystatin 100 100 100
Greseofulvin 500 100 100
Section-IV: Application
Section-IV: Application 191
Continue…
Dye No. Minimal fungicidal Concentration (µg/ml)
C. albicans A. niger A. clavatus
MTCC 227 MTCC 282 MTCC 1323
D21 500 500 500
D22 1000 1000 1000
D23 500 1000 >1000
D24 200 200 200
D25 62.5 200 200
D26 1000 1000 1000
D27 100 500 500
D28 500 500 500
D29 >1000 >1000 >1000
D30 >1000 >1000 >1000
D31 100 250 250
D32 250 1000 1000
D33 500 1000 >1000
D34 250 500 500
D35 100 200 200
D36 500 500 500
D37 200 200 250
D38 500 500 500
D39 1000 1000 1000
D40 250 >1000 >1000
Nystatin 100 100 100
Greseofulvin 500 100 100
Section-IV: Application
Section-IV: Application 192
Thermogravimetric Analysis (TGA)
The thermal degradation of dye depends upon various variables. Important
variables include decomposition temperature, heating rate, rate of removal of volatile
products from the reaction zone, softening and melting points of the dye, sample size
and dimensions, presence of oxygen in the atmosphere or absorbed in the sample or
other occluded impurities, the initiators used and the mechanism of termination and
many other factors.
In order to identify the generalized mechanism, therefore, thermal
decomposition is carried out under inert atmosphere with carefully purified reactive
dyes samples in the form of finely divided powder.
Thermogravimetric curves obtained at a heating rate of 10°C/min in nitrogen
(30 ml/min) atmosphere for reactive dyes are shown in Figure-69 to 76 and TGA data
of reactive dyes at various temperatures are given below.
Table-39: Temperature (ºC) characteristics of reactive dyes
Dyes No. % Weight loss at various temperature (ºC) from TGA
150 300 450 600 750 900
D1 3.85 5.89 9.49 11.75 16.45 29.01
D5 1.86 4.67 7.91 9.58 14.30 30.82
D11 2.02 4.22 6.52 9.02 11.52 32.52
D25 3.21 5.21 9.01 11.41 15.01 35.01
D21 2.82 5.12 8.12 10.52 15.02 32.52
D25 2.53 4.88 7.18 9.93 14.53 36.03
D31 1.01 4.11 6.11 8.01 14.01 33.76
D35 1.60 3.55 5.65 7.65 13.40 30.55
Thermal stability of the reactive dyes on the basis of initial decomposition
temperature 150ºC is as follow:
D1 > D25 > D21 > D25 > D11 > D5 > D35 > D31
Most of the reactive dyes degrade in a single step. All the samples start their
degradation at around 150ºC and weight loss of about 1.01-3.85% was observed. The
rate of degradation of all the samples increase rapidly between 750-900ºC and the
weight loss of about 29.01-36.03% was observed.
Section-IV: Application
Section-IV: Application 193
Figure-69: Thermogravimetric curves of dyes D1
-100 0 100 200 300 400 500 600 700 800 900 1000
70
75
80
85
90
95
100 PerkinElmer Thermal Analysis
Heat from 30.00oC to 900
oC at 10.00
oC/min
Wei
ght
(%)
Temperature (oC)
Figure-70: Thermogravimetric curves of dyes D5
-100 0 100 200 300 400 500 600 700 800 900 1000
70
75
80
85
90
95
100
Heat from 30.00oC to 900.00
oC at 10.00
oC/min
PerkinElmer Thermal Analysis
Wei
ght
(%)
Temperature (oC)
Section-IV: Application
Section-IV: Application 194
Figure-71: Thermogravimetric curves of dyes D11
-100 0 100 200 300 400 500 600 700 800 900 1000
65
70
75
80
85
90
95
100
105
Heat from 30.00oC to 900.00
oC at 10.00
oC/min
PerkinElmer Thermal Analysis
Wei
ght
(%)
Temperature (oC)
Figure-72: Thermogravimetric curves of dyes D15
-100 0 100 200 300 400 500 600 700 800 900 1000
60
65
70
75
80
85
90
95
100
105
Heat from 30.00oC to 900.00
oC at 10.00
oC/min
PerkinElmer Thermal Analysis
Wei
ght
(%)
Temperature (oC)
Section-IV: Application
Section-IV: Application 195
Figure-73: Thermogravimetric curves of dyes D21
-100 0 100 200 300 400 500 600 700 800 900 1000
65
70
75
80
85
90
95
100
105
Heat from 30.00oC to 900.00
oC at 10.00
oC/min
PerkinElmer Thermal Analysis
Wei
ght
(%)
Temperature (oc)
Figure-74: Thermogravimetric curves of dyes D25
-100 0 100 200 300 400 500 600 700 800 900 1000
60
65
70
75
80
85
90
95
100
105
Heat from 30.00oC to 900.00
oC at 10.00
oC/min
PerkinElmer Thermal Analysis
Wei
ght
(%)
Temperature (oC)
Section-IV: Application
Section-IV: Application 196
Figure-75: Thermogravimetric curves of dyes D31
-100 0 100 200 300 400 500 600 700 800 900 1000
65
70
75
80
85
90
95
100
105
Heat from 30.00oC to 900.00
oC at 10.00
oC/min
PerkinElmer Thermal Analysis
Wei
ght
(%)
Temperature (oC)
Figure-76: Thermogravimetric curves of dyes D35
-100 0 100 200 300 400 500 600 700 800 900 1000
65
70
75
80
85
90
95
100
Heat from 30.00oC to 900.00
oC at 10.00
oC/min
PerkinElmer Thermal Analysis
Wei
ght
(%)
Temperature (oC)
Section-IV: Application
Section-IV: Application 197
Overall Evaluation of Reactive Dyes
The main purpose of this investigation was to design and synthesis mono azo
reactive dyes containing a heterocyclic chromophoric moiety like quinazolinone
molecule in good yield. Percentage yields were fairly high for all dyes ranging from
78 to 85% (Table 1-4). This indicated a relatively high level of conversion from the
starting materials to synthesized dye. The dyes were applied to silk, wool and cotton
fabrics.
Three aspects are considered for overall evaluation,
(a) Shade,
(b) Dyeing properties,
(c) Fastness properties,
(d) Colorimetric data and
(e) Antimicrobial activity
Evaluation on silk, wool and cotton fibres
(a) Shade
Dyed pattern mounted on shade cards 1 & 2 for silk, 3 & 4 for wool and 5 & 6
for cotton shows that reactive dyes of the type Series-1, Series-2, Series-3 and
Series-4 produce purple to yellow shades. The shades are deeper in the case of silk
and wool than that of cotton fibre. This is probably due to the high substantivity of
the dyes on silk and wool fibres.
The variation in the hues depending on the nature and position of the electron
donating and electron attracting groups attached to the coupling components used, i.e.
dyes having H-acid as coupling component gives purple color hue, J-acid as coupling
component gives yellow color and N-phenyl-J-acid as coupling components gives red
color.
From the visualization of shades of all the dyes, we can arrange them by
decreasing order of depth of shade, which is as follows:
Series-2 > Series-1 > Series-3 > Series-4
It means that Series-2 has darker depth of shade than remaining three series and
Series-4 has lighter depth of shade than remaining three series.
Section-IV: Application
Section-IV: Application 198
(b) Dyeing properties
The results of the exhaustion study reveals that exhaustion on silk fibre ranges
from 84 to 91%; for wool fibre ranges from 84 to 92% and for cotton fibre ranges
from 82-91%.
The data regarding the fixation on the silk fibre ranges from 74-81%; for wool
fibre ranges from 75 to 83% and for cotton fibre ranges from 72 to 81% respectively.
This indicates that the dyes have good affinity and solubility with the fabric.
A remarkable degree of levelness after washing indicated good penetration and
excellent affinity to the fibers. The introduction of a s-triazine group to the dye
molecule improves the degree of exhaustion and fixation of the dyes.
(c) Fastness properties
All the reactive dyes show generally moderate to very good light fastness
property and good to excellent washing and rubbing fastness properties on silk, wool
and cotton fibres. This is attributed to good penetration and affinity of the reactive
dyes to the fibres. Good light fastness may be due to the higher stability of resonance
in the conjugated system and the dye structure within the fabric in an aggregated or
crystalline form being transformed from a dispersed form.
(d) Colorimetric data
Colorimetric data results for silk, wool and cotton fibres are as follows:
For silk fibre
All the dyes showed reddish-yellow shade except two dyes D21 and D31
showed reddish-blue shade with good lightness and brightness. Dye D11 showed
maximum color strength value (10.76) while D34 showed minimum color strength
value (0.25).
For wool fibre
All the dyes showed reddish-yellow shade with full of lightness and
brightness. Dye D11 showed maximum color strength value (26.18) while dye D17
showed minimum color strength value (2.05). The color yield values (K/S) seem to be
higher on wool in comparison with silk and cotton fibres. This is probably due to the
high substantivity of the dyes on wool fibre.
Section-IV: Application
Section-IV: Application 199
For cotton fibre
All the dyes showed reddish-yellow shades except two dyes D21 and D31
which showed reddish-blue shade with good lightness and brightness. Dye D11
showed maximum color strength value (13.30) while dye D17 showed minimum color
strength value (0.89).
The lower value of K/S may be due to the pronounced aggregation of the dye
molecule in the dye bath leading to the reduced solubility and prevention of the dye
molecule being absorbed on the fibre surface.
(e) Antimicrobial activity
All the synthesized reactive dyes samples (D1 to D40) were tested for their
antibacterial and antifungal activity (MIC) in vitro by broth dilution method with two
Gram positive bacteria S. aureus, S. pyogenes and two Gram negative bacteria E. coli,
P. aeruginosa and fungi C. albicans, A. niger and A. clavatus organisms taking
gentamycin, ampicillin, chloramphenicol, ciprofloxacin, norfloxacin, nystatin and
greseofulvin as standard drug.
Series-1:
Antibacterial activity
Dye D5 showed equipotential in activity against E. coli, P. aeruginosa and
S. pyogenus, while it showed excellent activity against S. aureus with respect to
standard drug Ampicillin. Dye D10 showed equipotential activity against E. coli and
S. aureus with respect to standard drug Ampicillin, Dye D7 showed euipotential
activity against E. coli; dyes D6 and D8 showed very good and good activity against
S. aureus with respect to standard drug Ampicillin.
Antifungal activity
Dyes D3 and D7 showed equipotential activity against C. albicans with respect
to standard drug Nystain, while dyes D1, D4, D5, D6 and D10 showed excellent activity
against C. albicans with respect to standard drug Greseofulvin. Dyes D2, D8 and D9
showed equipotential activity against C. albicans with respect to standard drug
Greseofulvin.
Section-IV: Application
Section-IV: Application 200
Series-2:
Antibacterial activity
Dye D11 showed excellent activity against E. coli and very good activity
against S. aureus with respect to standard drug Ampicillin. Dye D16 showed excellent
activity against E. coli and equipotential activity against S. aureus with respect to
standard drug Ampicillin and dye D13 showed equipotential activity against E. coli
and very good activity against S. aureus with respect to standard drug Ampicillin.
Antifungal activity
Dyes D14, D16 and D18 showed equipotential activity against C. albicans with
respect to standard drug Nystatin while dyes D11, D12 and D19 showed equipotential
acitivity against C. albicans with respect to standard drug Greseofulvin.
Series-3:
Antibacterial activity
Dye D25 showed equipotential activity against E. coli with respect to standard
drug Chloramphenicol and showed equipotential activity against P. aeruginosa and S.
aureus with respect to standard drug Ampicillin. Dye D29 showed excellent activity
against E. coli and equipotential activity against P. aeruginosa while very good
against S. aureus with respect to standard drug Ampicillin. Dye D22 showed
equipotential activity against E. coli and dye D30 showed equipotential activity
against P. aeruginosa while dye D21 showed excellent activity against S. aureus and
dye D24 showed equipotential activity against S. pyogenus with respect to standard
drug Ampicillin.
Antifungal activity
Dye D25 showed excellent activity against C. albicans and dye D27 showed
equipotential activity against C. albicans with respect to standard drug Nystatin. Dyes
D21, D23 and D28 showed equipotential activity and dye D24 showed very good activity
against C. albicans with respect to standard drug Greseofulvin.
Section-IV: Application
Section-IV: Application 201
Series-4:
Antibacterial activity
Dye D39 showed excellent activity against E. coli and equipotential activity
against P. aeruginosa while very good activity against S. aureus and good against S.
pyogenus with respect to standard drug Ampicillin. Dye D40 showed equipotential
activity against E. coli, P. aeruginosa and S. pyogenus with respect to standard drug
Ampicillin. Dyes D34 and D36 showed good activity against E. coli while dye D35
showed equipotential activity P. aeruginosa and very good against S. aureus. Dye D33
showed excellent activity against S. aureus, good against S. pyogenus while dye D34
showed excellent activity against S. aureus with respect to standard drug Ampicillin.
Antifungal activity
Dyes D31 and D35 showed equipotential activity against C. albicans with
respect to standard drug Nystatin. Dyes D32, D34, D37 and D40 showed excellent
activity against C. albicans while dyes D33, D36 and D38 showed equipotential activity
against C. albicans with respect to standard drug Greseofulvin.
The results demonstrate that certain dyes are able to reduce microbial growth
almost completely in the case of E. coli and C. albicans. Selected dyes would
therefore be valuable for the dyeing of sheets and gowns for hospital use, and on
articles which are less suitable for laundering such as mattresses and upholstery.
Thermogravimetric analysis
From the TGA analysis data it is found that the reactive dyes having good
thermal stability. Due to good thermal stability they can be applicable in high
technology application such as, liquid crystal display and color filters etc.
Section-IV: Application
Section-IV: Application 202
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