application of reactive dyes -...

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,



(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.



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.



(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.



(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



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.



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:



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



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.



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


dye, study of exhaustion of dye bath, study of the fixation of dye on the wool and the


(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


Glauber’s salt solution (20 % w/v) : 1.5 ml

Acetic acid solution (10 % w/v) : 1.5 ml

MLR : 1:40


pH of the dye bath : 3.0

Dyeing temperature : 100ºC


The steps involved are described below:



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


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.



(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



(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


dye, study of exhaustion of dye-bath, study of fixation of dye on the fabrics and the


(i) Application of Reactive Dyes to Cotton fabrics

Dyeing with all the reactive dyes to cotton fabric were carried out by the

following procedure:



Materials and conditions for 2% shade

Weight of the cotton fabric : 2.0 gm


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


pH of the dye bath : 8.0

Dyeing temperature : 100oC


This involves two steps:


(b) Dyeing


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


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



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


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



(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.



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




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


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


D31

D33

D35

D37

D39



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



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


(mg/ml)

D1

D3

D5

D7

D9


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


D11

D13

D15

D17

D19




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


(mg/ml)

D21

D23

D25

D27

D29


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


D31

D33

D35

D37

D39



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



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



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



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)




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



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




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)




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



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




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.



Table-32: Fastness properties of reactive dyes on wool fabrics

Dye

No.


No.

Fastness properties


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





Table-33: Fastness properties of reactive dyes on cotton fabrics:

Dye

No.


No.

Fastness properties


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





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.



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



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.



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



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



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



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



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



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



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)


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

)





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

)



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

)



Figure-49: Graph of K/S values of dyes D1-10 on wool fibre, which shows that dye D4


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

)





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

)



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

)



Figure-53: Graph of K/S values of dyes D1-10 on cotton fibre, which shows that dye


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)



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

)





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

)



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

)



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


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


-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




-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


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




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


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




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


-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


-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



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



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



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.



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



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.



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.



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.



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



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.



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



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



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



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



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.



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)


-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)




-100 0 100 200 300 400 500 600 700 800 900 1000

65

70

75

80

85

90

95

100

105


oC at 10.00

oC/min


Wei

ght

(%)

Temperature (oC)


-100 0 100 200 300 400 500 600 700 800 900 1000

60

65

70

75

80

85

90

95

100

105


oC at 10.00

oC/min


Wei

ght

(%)

Temperature (oC)




-100 0 100 200 300 400 500 600 700 800 900 1000

65

70

75

80

85

90

95

100

105


oC at 10.00

oC/min


Wei

ght

(%)

Temperature (oc)


-100 0 100 200 300 400 500 600 700 800 900 1000

60

65

70

75

80

85

90

95

100

105


oC at 10.00

oC/min


Wei

ght

(%)

Temperature (oC)




-100 0 100 200 300 400 500 600 700 800 900 1000

65

70

75

80

85

90

95

100

105


oC at 10.00

oC/min


Wei

ght

(%)

Temperature (oC)


-100 0 100 200 300 400 500 600 700 800 900 1000

65

70

75

80

85

90

95

100


oC at 10.00

oC/min


Wei

ght

(%)

Temperature (oC)



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.



(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.



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:


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.



Series-2:


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:


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.



Series-4:


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.



References

1. V. A. Shenai, Technology of Textile Procesing, Vol. VI, Technology of

Dyeing, 260 (1994).

2. T. Vickerstaff, J. Soc. Dyers Colors., 73 (1957) 237.

3. M. L. Gulrajani, Silk:Dyeing, Printing and Finishing, I. I. T., Delhi (1998).

4. E. R. Trotman, Dyeing and Chemical Technology of Textile Fibres, 4th ed.,

London, 590 (1970).


London, 333, 334 (1970).


London, 596, 597 (1970).


London, 589 (1970).

8. AATCC, 8 (1961).

9. C. M. Whittaker, C. C. Wilcock, Dyeing with Coal-tar Dyestuff, 6th

ed.,

Bolliere, Tindall and Cox, London, 23 (1964).

10. B. G. Skinner, T. Vickerstaff, J. Soc. Dyers Colors., 61 (1945) 193.

11. F. W. Billmeyer, Advance in Chemistry, Vol-107, Industrial Color

Technology, Ed., Robert F. Gould, ACS Publications (1971).

12. F. W. Billmeyer (Jr.), R. L. Abram, J. Paint Technol., 45 (1973) 310.

13. E. Allen, J. Optics Soc. Am., 56 (1966) 1256.

14. F. W. Billmeyer (Jr.), R. T. Mercus, Appl. Optics, 8 (1969) 763.

15. F. W. Billmeyer (Jr.), J. Coating Tech., 51 (1979) 46.

16. A. Rattan, Antimicrobials in laboratory medicine (2000) 85.

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