xerox mannual

8/19/2019 Xerox Mannual

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Cyclodextrins (CDs) are water soluble cyclic, non reducing oligosaccharides

consisting of D-(+)-gluocopyranose units linked through α-1,4-glycosidic linkages.

The major and most common three cyclodextrins, α-cyclodextrin, β-cyclodextrin and

γ-cyclodextrin consists of six, seven and eight D-(+)-glucopyranose units,

respectively. In CDs every gluocopyranose units has three free OH groups, two of

which (C-2 and C-3) are secondary, and one (C-6) is primary. Cyclodextrins are

modified for a wide variety of reasons. In CD derivative family, CD polymers reserve

a special position. In recent years attention has been expanding from supramolecular

chemistry of cyclodextrins to supramolecular chemistry of cyclodextrins based

polymers because more sophisticated structures and advanced functions have been

achieved by the formation of supramolecular cyclodextrin polymers. Compounds

consisting of covalently linked cyclodextrin rings are called cyclodextrin polymers.

The cyclodextrins fixed into polymeric structures behave differently from their

monomeric derivatives. CD polymers have the inclusion ability and the controlled

release ability, like their parent CDs. They have fine mechanical intensity and

excellent stability. Therefore, CD polymers have been studied for a long time and

received much attention in pharmaceutical industry, biomedical areas and

environmental chemistry.

Cyclodextrins (CDs) are water soluble: therefore they cannot be used as an adsorbent

in the uptake of dyes and various organic pollutants’. The only possible way is to

cross link the cyclodextrin either by using a suitable cross-linking agent or by

covalently binding the cyclodextrin to the backbone of existing polymers.

Chapter 3 deals with the synthesis of β-Cyclodextrin polymer using epichlorohydrin

as the cross-linker and also characterization by SEM and elemental analysis andremoval of some food dyes using β-Cyclodextrinepichlorohydrin polymer as a solid

phase extractant.


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3.1 PROCEDURE

3.1.1 Synthesis of β -cyclodextri nepichlorohydrin polymer

β-CDP was synthesized by method given in Literature (1a

). A brief procedure is given

here : 40g of β-CD, 10 g of soluble starch and 100 mL of 20% sodium hydroxide were

added into a beaker. The mixture was vigorously stirred at 50-60ºC until the reactants

dissolved. Total 60 mL of epichlorohydrin was added drop wise into the solution, and

β-CDP was formed in 30 min. Filtered with pressure through Buchner funnel and then

washed with distilled water 5-6 times, the polymer was dried at 100ºC and then stored

at room temperature in the desiccators for further use.


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Fig. 3.1.1 Represents generalized form showing cavity, toroidal form of β-

Cyclodextrin (β-CD) & β-Cyclodextrin polymer (β-CDP).

Fig. 3.1.2. Schematic representation of the Synthesis of β-

cyclodextrinepichlorohydrin polymer


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3.1.3 Characterization of β -CDP

The synthesized β-CD polymer was characterized by following techniques:

(i ) Scanning Electron Mi croscope (SEM) Analysis

(ii) FT-I R Analysis

3.1.3.1 Scanning El ectron M icroscope (SEM) Analysis

The synthesized polymer was subjected to SEM analysis to study their morphological

aspects. SEM Scans of the polymers were carried out at Sophisticated Instrumentation

centre (SIC), Punjabi University, Patiala on JEOL (Tokyo, JAPAN) JSM-6510/LV-

SEM Instrument. For the above experiment 1mg. of the sample was mounted on a

glass slide and subjected to scan at an acceleration voltage of 10Kv. The SEM images

were taken at five different magnifications from X200, X1000, X2500, X3000 and

X5000. From the SEM micrographs it is obvious that the structure of β-CD is rock

like and β-CDP is relatively fibrous which shows that cross-linking has taken place.


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Fig. 3.1.3 SEM scan of β-CD at X200 Fig. 3.1.4 SEM scan of β-CD at X1000

Fig. 3.1.5 SEM Scan of β-CD at X2500 Fig. 3.1.6 SEM Scan of β-CD at

X3000

Fig. 3.1.7 SEM Scan of β-CD at X5000


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Fig. 3.1.8 SEM scan of β-CDP at X100 Fig. 3.1.9 SEM scan of β-CDP at X1000

Fig. 3.1.10 SEM scan of β-CDP at X2500 Fig. 3.1.11 SEM scan of β-CDP at X3000

Fig. 3.1.12 SEM scan of β-CDP at X5000


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3.1.3.2 Fouri er Transform In fr a-red (FT-I R) Analysis

The reactant β-Cyclodextrin (β-CD), the synthesized β-Cyclodextrin polymer (β-

CDP) were subjected to IR analysis on a Pro resolution FTIR system as KBr pellets in

the range 4000-400 cm-1

region. From FT-IR (KBr) of β-CD, β-CDP it can be seen

that: for β-CD, 3600- 3000 cm-1 (OH), 2900 cm-1 (CH); while for β-CDP a sharp band

at 3400 cm-1 (OH), 3000 cm-1 (C-H stretch) and 1040 cm-1 (C-O-C stretch) is

obtained.


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Fig. 3.1.10 FT-IR Spectrum of native β-Cyclodextrin


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Fig. 3.1.11 FT-IR Spectrum of cross-linked β-Cyclodextrin polymer (β-CDP)


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3.2 DETERMINATION OF BRILLIANT GREEN DYE

Color [1] is a vital constituent of food and probably the first characteristic perceived

by the human senses. Food colorant is any substance that is added to food or drink to

change its color probably, for economic reasons. Brilliant green dye is a cationic and

triarylmethane dye of the malachite green series. This has been used in dyeing paper,

pulp, textile, plastics, leather, cosmetics and food (2). The present work describes β-

cyclodextrinepichlorohydrin polymer (β-CDP) used as a solid support for the

preconcentration and determination of the Brilliant green dye.

3.2.1 Material s

3.2.1.1 Equipment

A Shimadzu UV-1800 spectrophotometer (Shimadzu Ltd., Japan) equipped with the

matched 10-mm quartz cells was used to measure absorbance. Digital century pH-

meter CP-901 with a combined glass electrode was used to carry out pH

measurements. A thermostatic shaking water bath (Perfit India Ltd.) was used to carry

out all the inclusive procedures.

3.2.1.2 Reagents

All reagents used were of analR grade unless otherwise stated. Double distilled water

was used throughout the experiment. A 0.01M stock solution of Brilliant green dye

(Loba chem.) was prepared in double distilled water and further diluted as and when

required.

Buffer solution in the pH range of 1.0-3.5 were made by mixing equimolar solutions

of hydrochloric acid/Sodium Acetate and buffer solutions in the pH range of 4.0-6.5

were made by mixing 0.2 M sodium acetate and 0.2 M acetic acid solutions in the

different proportions. While those in the pH range of 7.0-11.0 were made by mixing

0.5 M ammonia and 0.5 M ammonium chloride.

The glass wares were washed with chromic acid and soaked in 5% nitric acid and then

cleaned with double distilled water before use and dried in an electric oven.


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3.2.2. Procedure

200 mg of β-CDP and 2.5 mL of buffer solution (pH 4.0) were added to a 100 ml

stoppered conical flask at room temperature. The mixture was allowed to stand for 15

min. so that β-CDP should swell sufficiently and an appropriate amount of dye was

added and made up to 90 ml with double distilled water. The mixture was shaken in

the thermostatic shaking water bath for 120 min. at the rate of 120 r.p.m. agitation

speed, 5.0 ml of supernatant solution was taken and the absorbance was measured

spectrophotometrically.

3.2.3. Optimization of various parameters

3.2.3.1 Ef fect of pH

The formation of inclusion complex of the dye in the polymer depends on the pH of

the sample solution which was studied in the range of (1.0-7.0.) using different buffer

solutions. The % uptake (≥ 95) was obtained at pH 4.0 (Fig. 3.2.3.1). Therefore, the

working pH was chosen as 4.0 for the subsequent studies.

3.2.3.2. Ef fect of shaking time

Shaking time is an important factor in determining the possibility of application of the

β-CD polymer for the selective uptake of brilliant green dye. Different shaking time

(ranging from 15 to 135 min.) were studied for the % uptake of brilliant green dye by

β-CD polymer. The results of Brilliant green dye show that the % uptake of (≥95%)

was attained within 120 min. (Fig. 3.2.3.2). Therefore, the shaking time of 120 min.

was selected for further studies.

3.2.3.3. Ef fect of sample volume

Enriching low concentration of dye from large volume of sample the effect of sample

volume is an important factor in determining the possibility of application of polymer

for the % of uptake of brilliant green dye. For this purpose 15, 30, 45, 60,75, 90 and

105 ml of sample volumes containing a fixed amount of dye were taken and uptake

of brilliant green dye were studied (Fig. 3.2.3.3). The maximum % uptake (≥95%) of

brilliant green dye was obtained at sample volume of 90 ml. Therefore, 90 ml of

sample volume was used for further studies.


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3.2.3.4. Ef fect of agi tation speed

Shaking speed is an important factor in determining the possibility of application of

polymer for the quantitative % uptake of brilliant green dye. The driving force i.e

Shaking speed could help in mass transfer and facilitate the concentration gradient

between the sample solution and the polymer. Different speeds (ranging from 40 to

140 r.p.m.) were studied for the % uptake of brilliant green dye by polymer. The

results of brilliant green dye uptake (Fig. 3.2.3.4) show that the % uptake reach

maximum (≥ 95%) at 120 r.p.m. Therefore, the shaking speed of 120 r.p.m. was

selected for further studies.

3.2.3.5. Ef fect of amount of polymer

The amount of the β-CD polymer is another important parameter that affects %

uptake of dye. A quantitative removal (≥ 95%) cannot be achieved when the β-CD

polymer is less than the optimum amount. In order to optimize the smallest amount of

polymer, 100 mg, 200 mg, 300 mg, 400 mg and 500 mg of the polymer were added to

the solution containing known amount of dye. The quantitative recoveries were

obtained at 200 mg of β-CDP (Fig. 3.2.3.5). Therefore, 200 mg of the β-CDP has been

used for further studies.


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Fig. 3.2.3.1. Effect of pH on the % uptake of the Brilliant green dye by the β-CD

polymer

Fig. 3.2.3.2. Effect of shaking time on the % uptake of the Brilliant green dye by the

β-CD polymer

Fig. 3.2.3.3. Effect of sample volume on the % uptake of the Brilliant green dye bythe β-CD polymer


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Fig. 3.2.3.4. Effect of agitation speed on the % uptake of the Brilliant green dye by

the β-CD polymer.

Fig. 3.2.3.5. Effect of amount of adsorbent on the % uptake of the Brilliant green by

β-CD the polymer.


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3.2.4. Appli cation

3.2.4.1 Determination of Br il li ant green dye in Mouth fr eshener

Green color tablets was grounded with a mortar and pestle, 3.0 g sample (mouthfreshener), added in double distilled water and was heated for some time to dissolve

the tablets properly and then cooled. After cooling double distilled water was added

to the sample and mixed thoroughly. The residue was filtered and filtrate was diluted

with double distilled water and made volume up to the mark in a standard flask. The

sample solution was then analyzed by the developed procedure. The results are given

in (Table 3.2.4.1.)

3.2.4.2 Determination of Br il li ant green dye in Jelly

5.0 g green color jelly was crushed and was heated for some time to dissolved

properly and then cooled. After cooling double distilled water was added to the

sample and mix thoroughly. The residue was filtered and filtrate was diluted with

double distilled water and made the volume up to the mark in the standard flask. The

sample solution was then analyzed by the developed procedure. The results are given

in (Table 3.2.4.2)

3.2.4.3 Determination of Br il li ant green dye in khus syrup

5 mL of the sample of khus syrup was transferred to a standard flask and made the

volume up to mark with double distilled water and solution was analyzed by

developed procedure. The results of analysis are given in (Table 3.2.4.2)


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Table 3.2.4.1 Results for the analysis of determination of Brilliant green dye in

Mouth Freshener.

Food Samples Added, μg/ml Found, μg/ml Recovery, %

aPizzle mouth

Freshener

0 0.012 _

0.268 0.266 95.00

0.536 0..536 97.76

1.072 1.072 99.25

aPizzle mouth Freshener - locally available in market

Table 3.2.4.2 Results for the analysis of determination of Brilliant green dye in Jelly

and Khus syrup.

Food samples Added, μg/ml Found, μg/ml Recovery, %

aJelly 0 0.025 -

0.268 0.267 95.35

0.536 0.536 97.38

1.072 1.072 99.19

bKhus syrup 0 0.014 -

0.268 0.270 96.42

0.536 0.536 95.69

1.072 1.088 95.53

aJelly - locally available in market, bKhus syrup – locally avaiable in market


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3.3 DETERMINATION OF SUDAN I DYE

Sudan I is a synthetic azo dye that is typically used in many industrial applications

including solvents, oils, fats, waxes, plastics, printing inks, and floor polishes (3,4).

The main reason for the widespread usage is its colorfastness and low price. Azo-

colorants are biologically active and have been associated with increased occurrence

of bladder cancer in textile and leather dyers, painters, and hairdressers (5-8). Due to

their potential carcinogenicity, many countries have banned the use of most azo dyes

at any level in products for human consumption. The present work describes β-

cyclodextrinepichlorohydr in polymer (β-CDP) as a solid support for the

preconcentration and determination of the Sudan I dye.

3.3.1 Material s

3.3.1.1 Equipment

Equipments are same as described in section 3.2.1.1

3.3.1.2 Reagents


was used throughout the experiment. A 0.01M stock solution of Sudan I dye (Loba

chem.) was prepared in double distilled water and further diluted as and when

required.

Buffer solutions are same as described in section 3.2.1.2

3.3.2 Procedure



min. so that β-CDP should swell sufficiently and appropriate amount of dye was


the thermostatic shaking water bath for 90 min. at the rate of 100 r.p.m., 5.0 ml of

supernatant solution was transferred into a test tube and the absorbance was measured

spectrophotometrically.


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3.3.3 Optimization of various parameters


The formation of inclusion complex of the dye in the polymer depends on the pH ofthe sample solution which was studied in the range of (1.0-7.0) using different buffer

solutions, % uptake (≥ 95) was obtained at pH 3.0. (Fig.3.3.3.1). Therefore, the


3.3.3.2. Ef fect of the shaki ng time

Shaking time is an important factor in determining the possibility of

application of the β-CD polymer for the selective uptake of Sudan I dye. Differentshaking time (ranging from 15 to 120 min) were studied for the % uptake of Sudan I

dye by β-CD polymer. The results of % uptake of Sudan I dye vs. the shaking time

show that the % uptake of (≥95%) was attained in 90 min.(Fig. 3.3.3.2). Therefore,

the shaking time of 90 min. was selected for further studies.

3.3.3.3 Ef fect of sample volume

Enriching low concentration of dye from large volume of sample the effect of samplevolume is an important factor in determining the possibility of application of

polymer for the % of uptake of Sudan I dye.For this purpose 15, 30, 45, 60, 75 and 90

mL of sample volumes containing a fixed amount of dye were taken and uptake

of Sudan I dye was studied (Fig. 3.3.3.3.). The maximum % uptake (≥95%) of

Sudan I dye was obtained at sample volume of 75 mL. Therefore, 75 mL of sample

volume was used for further studies

3.3.3.4 Ef fect of agitation speed

Speed of shaking is an important factor in determining the possibility of

application of polymer for the quantitative % uptake of Sudan I dye. The driving

force i.e Shaking speed could help in mass transfer and facilitate the concentration

gradient between the sample solution and the polymer. Different speeds (ranging from

40 to 140 r.p.m) were studied for the % uptake of Sudan I dye by the polymer. The

results of % uptake of Sudan I vs. agitation speed (Fig. 3.3.3.4) show that the %


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uptake reach maximum (≥95%) at 100 r.p.m. Therefore, the shaking speed of 100

r.p.m. was selected for further studies.

3.3.3.5. Ef fect of amount of polymer


uptake of dye. A quantitative removal (≥ 95%) cannot be achieved when the β-CD




obtained at 200 mg of β-CDP (Fig. 3.3.3.5). Therefore, 200 mg of the β-CDP has been



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Fig. 3.3.3.1. Effect of the pH on the % uptake of the Sudan I dye by β-CD polymer

Fig. 3.3.3.2.Effect of shaking time on the % uptake of the Sudan I dye by β-CD

polymer

Fig. 3.3.3.3. Effect of sample volume on the % uptake of the Sudan I dye by β-CD

polymer


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Fig. 3.3.3.4. Effect of agitation speed on the % uptake of the Sudan I dye by β-CD

polymer.

Fig. 3.3.3.5.Effect of amount of adsorbent on the % uptake of the Sudan I dye by β-

CD polymer


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Table 3.3.4.1. Results for the analysis of determination of Sudan I dye by developed

method in Chili Powder / Chili Sauce samples.

Food Samples Added, μg/mL Found, μg/mL Recovery, %

aChilli powder 0 0.017 -

0.331 0.334 95.97

0.662 0.662 97.49

0.892 0.890 97.90

bChilli Sauce 0 0.015 -

0.331 0.332 95.95

0.662 0.650 96.01

0.892 0.895 98.45

aChilli powder - locally available in market,

bChilli Sauce - locally available in market

Table 3.3.4.2. Results for the analysis of determination of Sudan I dye by developed

method in Tandoori masala mix sample.


aTandoori masala

mix

0 0.006 -

0.350 0.348 97.75

0.700 0.698 98.86

0.932 0.930 99.14

aTandoori masala mix – locally available in market


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3.4. DETERMINATION OF SUDAN III DYE

The most widely used synthetic organic azo dyes have been studied for their toxicity

risk (9). The chromophoric azo group on reduction forms suspected carcinogenic

aromatic amines under certain conditions (10). The Sudan III dye was used in food

industry with the regulation of maximum permissible levels in a particular foodstuff

(11). Based on toxicity data, various azo dyes are unauthorized and are sometimes

illegally used in food preparations either to enhance or to maintain the appearance of

food products (12-14). The adulteration of hot chili products with Sudan I, II, III and

IV led the EU to adopt emergency measure (15). The present work describes the use

of β-cyclodextrinepichlorohydr in polymer (β-CDP) as a solid support for the

preconcentration and determination of the Sudan III dye.

3.4.1 Material s

3.4.1.1 Equipment


3.4.1.2 Reagents


was used throughout the experiment. A 0.01M stock solution of Sudan III dye (Loba


required.


3.4.2 Procedure





the thermostatic shaking water bath for 75 min. at a rate of 120 r.p.m. agitation speed.

5.0 ml of supernatant solution was transferred into a test tube and the absorbance was

measured spectrophotometrically.


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The formation of inclusion complex of the dye in the polymer depends on the pH ofthe sample solution which was studied in the range of (1.0-7.0) using different buffer

solutions. % uptake (≥ 95) was obtained at pH 4.0. (Fig.3.4.3.1). Therefore, the


3.4.3.2 Ef fect of the shaking time


application of the β-CD polymer for the selective uptake of Sudan III dye. Differentshaking time (ranging from 15 to 120 min) were studied for the % uptake of Sudan

III dye by β-CD polymer. The results of % uptake of Sudan III dye vs. the shaking

time show that the % uptake of (≥95%) was attained within 75 min. (Fig. 3.4.3.2).

Therefore, the shaking time of 75 min. was selected for further studies.


Enriching low concentration of dye from large volume of sample the effect of samplevolume is an important factor in determining the possibility of application of

polymer for the % uptake of Sudan III dye. For this purpose 15, 30, 45, 60, 75 and 90

mL of sample volumes containing a fixed amount of dye were taken and uptake

of Sudan III dye was studied (Fig. 3.4.3.3). The maximum % uptake (≥95%) of

Sudan III dye was found with sample volume of 75 mL. Therefore, 75 mL of sample

volume was used for further studies

3.4.3.4 Ef fect of agitation speed

Shaking speed is an important factor in determining the possibility of application

of polymer for the quantitative % uptake of Sudan III dye. The driving force i.e



140 r.p.m) were studied for the % uptake of Sudan III dye by polymer. The results of

% uptake of Sudan III vs. agitation speed (Fig. 3.4.3.4) shows that the % uptake was


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Fig. 3.4.3.1. Effect of the pH on the % uptake of the Sudan III dye by β-CD polymer

Fig. 3.4.3.2.Effect of shaking time on the % uptake of the Sudan III dye by β-CD

polymer

Fig. 3.4.3.3 Effect of sample volume on the % uptake of the Sudan III dye by β-CD

polymer


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Fig. 3.4.3.4. Effect of agitation speed on the % uptake of the Sudan III dye by β-CD

polymer

Fig. 3.4.3.5. Effect of amount of adsorbent on the % uptake of the Sudan III dye by

the β-CD polymer


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3.4.4. Appl ications

3.4.4.1. Determination of Sudan I I I in Curr y Powder/ Curry Paste

3.0 g of the Curry powder/ curry paste sample was dissolved in hot double distilledwater. After mixing, the residue was filtered, and filtrate was diluted with double

distilled water and made the volume up to the mark in a standard flask. The sample

solution was then analyzed by the developed procedure. The results of analysis are

given in (Table 3.4.4.1)

3.4.4.2. Determination of Sudan I I I in Navratan oil

5.0 mL of Navratan oil, dissolved in Dimethylformamide (DMF). After mixing, theresidue was filtered, and filtrate was diluted with double distilled water and made the

volume up to mark in a standard flask. The sample solution was then analyzed by the

developed procedure. The results of analysis are given in (Table 3.4.4.2).


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Table 3.4.4.1 Results for the analysis of determination of Sudan III dye by developed

method in Curry powder and Curry paste sample.


aCurry powder 0 0.015 -

0.254 0.256 95.16

0.440 0.440 96.70

0.920 0.918 98.18

bCurry paste 0 .010 -

0.260 0.262 97.03

0.450 0.452 98.26

0.950 0.948 98.75

aCurry powder – Locally available in market bCurry paste – locally available in

marke

Table 3.4.4.2 Results for the analysis of determination of Sudan III dye by developed

method in some Navratan cosmetic samples.


a Navratan oil 0 0.019 -

0.274 0.286 97.61

0.469 0.470 96.31

0.939 0.950 99.16

a Navratan oil – locally available in market


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3.5 DETERMINATION OF SUDAN IV DYE

Sudan dyes are a class of lipophilic synthetic organic colorants that are widely used

in industry because of their colorfastness and low price. These azo-dyes are known to

be biologically active through their metabolites (16), and they represent a potential

risk to public health if they enter the food chain. Sudan IV dye have been associated

with the occurrence of bladder cancer in textile and leather dyers, painters, and hair-

dressers (17,18). Sudan IV is considered a possible human carcinogen and mutagen

(19), classified as a category 3 carcinogen by the International Agency for Research

on Cancer (IARC) (20). Sudan IV dye has been found to cause a high frequency of

bladder carcinomas in mice (21). The present work describes β-

cyclodextrinepichlor ohydrin polymer (β-CDP) as a solid support for the

preconcentration and determination of the Sudan IV dye.

3.5.1 Material s

3.5.1 Equipment


3.5.1.2 Reagents


was used throughout the experiment. A 0.01M stock solution of Sudan IV dye (Loba


required.


3.5.2 Procedure





the thermostatic shaking water bath for 120 min. at the rate of 120 r.p.m. agitation


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speed, 5.0 ml of supernatant solution was transferred into a test tube and the

absorbance was measured spectrophotometrically.

3.5.3 Optimization of various parameters


The formation of inclusion complex of the dye in the polymer depends on the pH of

the sample solution which was studied in the range of (1.0-7.0) using different buffer

solutions (Fig. 3.5.3.1), % uptake (≥95) was obtained at pH 3.0. Therefore, the


3.5.3.2 Eff ect of the shaki ng time


application of the β-CD polymer for the selective uptake of Sudan IV dye. Different

shaking time (ranging from 15 to 135 min) were studied for the % uptake of Sudan

IV dye by β-CD polymer. The results of % uptake of Sudan IV dye vs. the shaking

time show that the % uptake of (≥95%) was attained within 120 min.(Fig. 3.5.3.2).



Enriching low concentration of dye from large volume of sample the effect of sample

volume is an important factor in determining the possibility of application of

polymer for the % of uptake of Sudan IV dye.For this purpose 15, 30, 45, 60, 90 and

105 mL of sample volumes containing a fixed amount of dye were taken and

uptake of Sudan IV dye was studied (Fig. 3.5.3.3). The maximum % uptake

(≥95%) of Sudan IV dye was obtained at sample volume of 90 mL. Therefore, 90 mL

of sample volume was used for the further studies.

3.5.3.4.Ef fect of agitation speed

Shaking speed is an important factor in determining the possibility of application

of polymer for the quantitative %uptake of Sudan IV dye. The driving force i.e




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140 r.p.m) were studied for the % uptake of Sudan IV dye by polymer. The results of

% uptake of Sudan IV vs. agitation speed (Fig. 3.5.3.4) show that the % uptake reach

maximum (≥95%) at 120 r.p.m. Therefore, the shaking speed of 120 r.p.m. was

selected for further studies.

3.5.3.5 Ef fect of amount of polymer


uptake of dye. A quantitative removal (≥95%) cannot be achieved when the β-CD




obtained at 200 mg of β-CDP (Fig. 3.5.3.5). Therefore, 200 mg of the β-CDP was



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Fig. 3.5.3.1. Effect of the pH on the % uptake of the Sudan IV dye by β-CD polymer

Fig. 3.5.3.2. Effect of shaking time on the % uptake of the Sudan IV dye by the β-CD

polymer

Fig. 3.5.3.3. Effect of sample volume on the % uptake of the Sudan IV by the β-CD

polymer


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Fig. 3.5.3.4. Effect of agitation speed on the % uptake of the Sudan IV dye by the β-

CD polymer

Fig. 3.5.3.5. Effect of amount of adsorbent on the % uptake of the Sudan IV dye by

the β-CD polymer


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3.5.4.1. Determination of Sudan I V i n H ot Chi li Tomato Sauce/ Chi li pepper

3.0 g of the Hot chili tomato sauce/ Chili pepper sample was dissolved in hot doubledistilled water. After mixing, the residue was filtered, and filtrate was diluted with

double distilled water and made the volume up to the mark in a standard flask. The

sample solution was then analyzed by the developed procedure. The results of

analysis are given in (Table 3.5.4.1)

3.5.4.2. Determination of Sudan I V i n Red Sausage

3.0 g of the Red Sausage sample was dissolved in hot double distilled water. Aftermixing, the residue was filtered, and filtrate was diluted with double distilled water

and made the volume up to the mark in a standard flask. The sample solution was then

analyzed by the developed procedure. The results of analysis are given in (Table

3.5.4.2)


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Table 3.5.4.1. Results for the analysis of determination of Sudan IV dye by developed

method in Hot Chilli Tamoto Sauce and Hot Chilli Pepper food samples.


aHot Chilli Tamoto

Sauce

0 0.012 -

0.258 0.260 96.29

0.550 0.548 97.50

0.936 0.930 98.10

bHot Chilli Pepper 0 0.014 -

0.258 0.264 97.05

0.550 0.552 98.87

0.936 0.938 97.73

aHot Chilli Tamoto Sauce - locally available in market, bHot Chilli Pepper - locally

available in market

Table 3.5.4.2 Results for the analysis of determination of Sudan IV dye by developed

method in Red Sausages sample.


aRed Sausage 0 0.011 -

0.260 0.262 97.03

0.568 0.570 98.44

0.945 0.950 99.37

aRed Sausage – locally available in market


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3.6 DETERMINATION OF SUNSET YELLOW DYE

Synthetic dyes are widely used for improving the color and enhancing the visual

aesthetic appeal of some foods, and this effect is maintained throughout the

production process and during storage. They present high stability to light, oxygen,

and pH changes, and have lower prices compared to natural dyes (22-23). These dyes

are added as small amounts in food, drugs or cosmetic products (24-26). Sunset

yellow is widely used as additives in soft drinks and other non-alcoholic drinks, the

highest dose of each in soft drinks being 70.0 mg 1 -1 (27). The present work describes

that β-cyclodextrinepichlorohydr in polymer (β-CDP) as a solid support for the

preconcentration and determination of the Sunset yellow dye.

3.6.1 Material s

3.6.1.1 Equipment


3.6.1.2 Reagents


was used throughout the experiment. A 0.01M stock solution of Sunset Yellow dye

(Loba chem.) was prepared in double distilled water and further diluted as and when

required.


3.6.2. Procedure





the thermostatic shaking water bath for 90 min., at the rate of 140 r.p.m., 5.0 ml of

supernatant solution was transferred into a test tube and the absorbance was measured

spectrophotometrically


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The formation of inclusion complex of the dye in the polymer depends on the pH ofthe sample solution which was studied in the range of (1.0-7.0.) using different buffer

solutions (Fig. 3.6.3.1), % uptake (≥ 95) was obtained at pH 3.0. Therefore, the


3.6.3.2 Eff ect of shaking time

Shaking time is an important factor in determining the possibility of application of the

β-CD polymer for the selective uptake of sunset yellow dye. Different shaking time(ranging from 15 to 105 min.) were studied for the % uptake of sunset yellow dye by

β-CD polymer. The results of % uptake of Sunset yellow dye vs. the shaking time

show that the % uptake of (≥95%) was attained within 90 min.(Fig. 3.6.3.2).



Enriching low concentration of dye from large volume of sample the effect of samplevolume is an important factor in determining the possibility of application of polymer

for the % of uptake of sunset yellow dye. For this purpose 15, 30, 45, 60, 75 and 90

ml of sample volumes containing a fixed amount of dye were taken and uptake of

sunset yellow dye was studied (Fig. 3.6.3.3). The maximum % uptake (≥95%) of

sunset yellow dye was at sample volume of 75ml. Therefore, 75ml of sample volume

was used for the further studies.

3.6.3.4. Ef fect of agi tation speed

Shaking speed is an important factor in determining the possibility of application of

polymer for the quantitative % uptake of sunset yellow dye. The driving force i.e



140 r.p.m.) were studied for the % uptake of Sunset yellow dye by polymer. The

results of % uptake of sunset yellow vs. agitation speed (Fig. 3.6.3.4) shows that the


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% uptake reach maximum (≥ 95%) at 140 r.p.m. Therefore, the shaking speed of 140

r.p.m. was selected for further studies.

3.6.3.5 Ef fect of amount of polymer

The amount of the β-CD polymer is another important parameter that affects %uptake

of dye. A quantitative removal (≥ 95%) cannot be achieved when the β-CD polymer is

less than the optimum amount. In order to optimize the smallest amount of polymer,

100mg, 200mg, 300mg, and 400mg of the polymer were added to the solution

containing known amount of dye. The quantitative recoveries were obtained at 200

mg of β-CDP shown in (Fig. 3.6.3.5). Therefore, 200 mg of the β-CDP has been used

for further studies.


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Fig. 3.6.3.1. Effect of pH on the % uptake of the Sunset yellow dye by the β-CD

polymer

Fig. 3.6.3.2. Effect of shaking time on the % uptake of the Sunset yellow dye by the

β-CD polymer

Fig. 3.6.3.3. Effect of sample volume on the % uptake of the Sunset yellow dye by

the β-CD polymer


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Fig. 3.6.3.4. Effect of agitation speed on the % uptake of the Sunset yellow dye by

the polymer

Fig. 3.6.3.5 Effect of amount of adsorbent on the % uptake of the Sunset yellow dye

by the β-CD polymer


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3.6.4.1. Determination of Sunset yell ow in Santr a Goli (orange f lavor)

8.0 g sample of Santra goli was grounded in mortar with pestle and then dissolved inhot double distilled water. The solution was cooled and filtered. Filtrate was diluted

with double distilled water and made the volume up to the mark in a standard flask.

The sample solution was then analyzed by the developed procedure. The results of

analysis are given in (Table 3.6.4.1)

3.6.4.2. Determination of Sunset yell ow in M ir inda (orange fl avor)

5.0 mL of Mirinda sample was dissolved in double distilled water. After mixing, theresidue was filtered, and filtrate was diluted with double distilled water and made the

volume up to the mark in a standard flask. The sample solution was then analyzed by

the developed procedure. The results of analysis are given in (Table 3.6.4.1).

3.6.4.3. Determination of sunset yell ow in Dry Syrup (ZI F I 100)

5.0 mL of Dry syrup sample was dissolved in double distilled water. After mixing, the

residue was filtered, and filtrate was diluted with double distilled water and made thevolume up to the mark in a standard flask. The sample solution was then analyzed by

the developed procedure. The results of analysis are given in (Table 3.6.4.2).


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Table 3.6.4.1 Results for the analysis of determination of Sunset Yellow dye by

developed method in Santra Goli and Mirinda food samples.

Food Samples Added, µg/ml Found, µg/ml Recovery, %

aSantra Goli 0 0.010 -

0.304 0.302 96.17

0.603 0.590 96.24

1.206 1.208 99.34

bMirinda 0 0.022 -

0.304 0.301 95.85

0.603 0.595 95.20

1.206 1.201 97.80

aSantra goli - locally available in market,

bMirinda - locally available in market

Table 3.6.4.2 Results for the analysis of determination of Sunset yellow dye by

developed method in Dry Syrup (ZIFI 100) pharmaceutical sample.

Food Samples Added, µg/ml Found, µg/ml Recovery, %

aDry Syrup (ZIFI

100)

0 0.011 -

0.228 0.232 97.07

0.454 0.458 98.49

0.808 0.810 98.90

a

Dry Syrup – locally available in market


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113

Table 3.7 SHOWING OPTIMIZED CONDITIONS FOR THE

DETERMINATION OF FOOD DYES USING β-CDEPICHLOROHYDRIN

AS SOLID PHASE EXTRACTANT

Food dyes pH Shaking

time (min.)

Sample

volume (mL)

Agitation

speed (r.p.m.)

Amount of

adsorbent (mg)

Brilliant

green

4.0 120 90 120 200

Sudan I 3.0 90 75 100 200

Sudan III 4.0 75 75 120 300

Sudan IV 3.0 120 90 120 200

Sunset

yellow

3.0 90 75 140 200


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