STRATEGIES FOR STABILIZATION OF

NATURAL COLORANTS

An Investigation

By

RAHUL SHARMA

Submitted in partial fulfillment for the Degree of

Master of Science in Food Technology

University of Mysore
Mysore, Karnataka

Human Resource Development Department

Central Food Technological Research Institute
Mysore–570020

JUNE 2005
 Acknowledgement
It gives me great pleasure to express my heartfelt thanks and gratitude to my guide Dr. P. Srinivas,
Scientist, Plantation Products, Spices and Flavor Technology Department, Central Food
Technological Research Institute, Mysore for his valuable guidance, worthy suggestions and
continuous encouragement throughout the investigation work.

I am extremely grateful to Dr. V. Prakash, Director, CFTRI, Mysore, Mr. B. Raghvan, Head,
PPSFT Department and Dr. M. C. Varadaraj, Head, Human Resource Development for providing
me an opportunity to undertake this work.

I wish to extend my thanks to Mr. S. R. Sampathu, Scientist, PPSFT, Ms. H. B. Sowbhagya,

Scientist, Ms. K. Ramalakshmi, Scientist PPSFT and Mr. S. G. Jayaprakash, FEPD for guidance
and help during the product preparation and color assay.

I would like to extend my thanks to Dr. B. S. Jena, Dr. R. P. Singh, Dr. P. S. Negi & Dr. G. K.
Jayaprakash of HRD for their constant encouragement and all other staff members of HRD for
their cooperation.

I am also thankful to Dr. H. H. Pattekhan, Scientist, TTBD and Mr. J. R. Manjunath and Ms.
Divya Parvati, research students in PPSFT for their assistance during the experimental work in the
laboratory. I convey my sincere thanks to faculty members of the PPSFT department for their help
during the course of this work.

I am highly gratified to my beloved parents, brothers and sisters for their constant enlightenment,
encouragement and love.

My thanks are also due to all my friends and classmates, who are associated with the successful
completion of this work.

CFTRI, Mysore
Date: 21 June 2005 Rahul Sharma
Dr. P. Srinivas, 21 June 2005
Scientist,
Plantation Products, Spices and Flavor Technology,
Central Food Technological Research Institute,
Mysore.

CERTIFICATE

I certify that this investigation report on ‘Strategies for Stabilization of Natural Colorants’
submitted by Mr. Rahul Sharma in partial fulfilment of the award of Degree of Master of Science in
Food Technology of the University of Mysore, is written by him based on the investigation work
carried out under my guidance in the Department of Plantation Products, Spices and Flavor
Technology, Central Food Technological Research Institute, Mysore.

(Dr. P. Srinivas)
 INDEX

SR. No. CHAPTER PAGE NO.

1. INTRODUCTION 1

2. MATERIALS AND METHODS 10

3. RESULT AND DISCUSSIONS 14

4. SUMMARY AND CONCLUSIONS 21

BIBLIOGRAPHY 23
 Chapter One

CHAPTER 1

INTRODUCTION

General

The color of foodstuff is an important attribute, which is considered as one of the

primary characteristics perceived by the senses that helps in determining the
acceptability, judging the quality and appreciating the aesthetic value of a foodstuff
(Sampathu et al., 1981; Santhanakrishna et al, 1981; and Pattnaik et al, 1999). Color
helps in recognition of food and also makes it appealing to the consumer. The consumer
tends to judge the quality of a food product by its color. Also, appetite is stimulated or
dampened in almost direct relation to the observer’s reaction to color. The color,
therefore, has a direct bearing on the flavor of a food product. In terms of food
manufacturing a good strawberry color, for example, can indicate to the consumer a high
quality product, whilst a washed out or artificially bright product can indicate poor
quality or an inferior product.

The practice of adding colors to processed foodstuffs to make them attractive dates
back to very ancient times. Our ancestors used saffron, turmeric and vegetable dyes. In
1856 the first synthetic color (mauvine), was developed by Sir William Henry Perkin
(Walford, 1980) and by the turn of century, unmonitored color additives had spread
through USA and Europe in many popular foods, including ketchup, mustard, jellies and
wine. Sellers at that time offered more than 80 artificial coloring agents, as these were
found more technically suitable for use in food than natural ones in efficiency, shades,
range and cost. Thus for many years synthetic colorants were used in majority of
formulations without having been tested for their safety. From 1907, certification
requirements saw many of them gradually withdrawn, as they were found harmful. The
list of certified synthetic colorants has been continuously shrinking since then. In India, a

1
 Chapter One

blanket ban on use of synthetic color additives has been suggested in 1981
(Sankarnarayan, 1981).

In recent years, therefore, manufacturers have shown a greater interest in the

natural colorants. Identification of alternative safe colorants from natural sources and
development of a few synthetic routes to then have partly met the color need of food
industry. Also due to public prejudice against synthetic additives, manufacturers want to
label their product as having, a natural ingredient or no additives and hence, the
preference to use the natural colorants.

Why is Color added:

The colors are added to the foodstuff on the basis of some specific reasons (Sampathu et
al, 1981), which are further governed by individual requirements. These reasons are as
follows:
1. Original appearance:

Due to the effect of various processing conditions, the natural color of foodstuff
gets destroyed e.g. color of canned pear, beans, strawberries, etc. is lost during
storage. To restore the original appearance of these foods, it is necessary to add color
in the foodstuffs.

2. Uniformity of Product:

Product obtained may vary in their appearance from season to season and even
batch-to-batch e.g. addition of a specific quantity of color becomes necessary e.g. in
case of butter, to make the product uniform throughout the year.

3. Intensifying Natural Color:

In some cases, the colors of the natural ingredients are too weak and it is well
known that the consumer prefers stronger colors. So colors are used to intensify colors
of those naturally occurring in foods e.g. fruit yogurts.

2
 Chapter One

4. Product Identification:

There are some products where there is little or no color of its own e.g. liquid
beverages, sherbets, jams, and jellies. So color addition helps to preserve the identity
or character of the food.

5. Protection:

Coloring helps to protect flavor and light sensitive vitamins during shelf storage
by a sunscreen effect.

6. Attractive Appearance:

To give an attractive appearance to food, which would otherwise look unattractive

or unappetizing, e.g. colorless gelatin base jelly and thus enhance enjoyment.

7. Antioxidant and nutroceutical action

The carotenoids and other pigments like curcumin have been shown to possess
strong antioxidant properties and a host of other health promoting and disease
alleviation effects which further enhance the importance of incorporation and use of
natural colours in foods and other products.

Some other benefits of addition of colorants to food products are as follows

(Pattnaik et al, 1999):

1. Some indication for how much to use and how long to store.
2. Improved stability and reduced wastage in storage.
3. Reduced cost and improved quality through manufacturer benefits.

3
 Chapter One

Market Trends:

There are no reliable published statistics on the size of the indigenous color
market. However, on a global scale a reasonable estimate would be $940 million, which
can be segmented as follows (Downham, 2000):

a) Synthetic colors - $400 million

b) Natural colors - $250 million (of which $100 million is in the USA)

c) Nature identical colors - $189 million

d) Caramel colors - $100 million

Consumer pressure, sociological changes and technological advances leading to

more advances in the food processing industry have increased the overall color market.
The most significant growth has been in naturally derived colors owing to the
improvements in stability as well as the food industries aim to meet the increasing
consumer perception that ‘natural is the best’. Future growth is thought to be going to be
greatest for naturally derived colors with a predicted annual growth rate of 5-10%.
Synthetic colors are still forecast to grow but a lower rate of between 3 and 5%.

Natural Colors:

Natural colors are the colors extracted from natural sources or manufactured by
biosynthetic routes. Natural colour is spread widely throughout nature in fruits,
vegetable, seeds and roots. In our daily diets we consume large quantity of many
pigments especially anthocyanins, carotenoids and chlorophylls. These are also known as
naturally derived colors. The shades and key characteristics of most commonly used
permitted naturally derive colour are shown in table 1.0 (Downham and Collins, 2000).

4
 Chapter One

Table – 1

Pigment Sources Color Shade Comments

Curcumin Turmeric Bright lemon A non-soluble pigment that is light
Rhizome Yellow sensitive once solubilized and in
roots the presence of water. Used widely
in desserts and confectionery.
Lutein Marigold Golden Oil soluble carotenoid pigment
Alfalfa Yellow naturally present to use water-
soluble forms with increased
oxidative stability has extended its
use to applications such as desserts,
soft drinks and bakery products.
Only permitted in the USA in
chicken feed.
Natural Palm oil, Golden Another oil soluble mixture of
mixed D. Salina yellow to carotenioids (principally β-
carotenes (algae) orange carotene), which is naturally
sensitive to oxidation. The USA
permits only algal source. Has pro
vitamin A activity
Bixin/ Bixa orellana Orange Bixin is oil soluble and norbixin
Norbixin bush seeds water dispersible. Widely used in
ice cream, cheese, yellow fats and
smoked fish
Capsanthin/ Paprika Reddish Natural oil soluble carotenoids.
Capsorubin Capsicum orange Used in savory products such as
annum L. sauces and coatings as well as
desserts, confectionery and bakery
products.
Lycopene Tomatoes Orange red Natural oil soluble carotenoid
pigment. Has found little
commercialization as a color owing
to high cost, poor stability and
orange rather than red shade.
Anthocyanin Black Pink/red to Natural pigments of many red
grapeskin, mauve/blue fruits, flowers and vegetables.
elderberries, depending on
black carrots, pH
red cabbage
Chlorophyll Grass, Olive green Chlorophylls are oil dispersible and
lucerne and chlorophyllins water-soluble.
nettle Limited use owing to poor stability
and dull color shade.

5
 Chapter One

However, these pigments from nature vary widely in their physical and chemical
properties. Many are sensitive to oxidation, pH change and light. Due to less stability of
natural colorant, this poses many problems in their use in food systems.

Strategies for Stabilization:

From last few past decades much emphasis has been given to colour formulation,
which offer stability to natural and natural identical pigments as they have much variation
in their solubility and stability to various conditions. Colour formulation development
has focus on improvement in three following areas.

1. Making water dispersible from natural oil soluble pigments by forming a dispersion
of oil soluble pigment throughout in aqueous disperse phase using emulsifier and
stabilizer.

2. Enhancing stability to oxidation via the synergistic use of antioxidant such as

tocopherol and ascorbyl palmitate.

3. Improved stability and clarity for application such as soft drink, jellies and
conserves.

Another approach to stability and solubility of natural colours is through

processing technology such as pigment encapsulation into insoluble carrier to produce
ranges of suspension with following benefits.

1. Improvement to light stability (e.g. Curcumin) and extending its area of

applications to product like sauces, dressing and soft drink.

2. Conversion of oil dispersible form of natural colours to water dispersible pigments

3. A reduced level of certain additives

6
 Chapter One

4. Production of different colour shades from those normally exhibited by the soluble
pigments

5. Less migration of colors during application such as in desserts.

In addition to this, protection provided is dependent on encapsulating matrix,

efficiency of encapsulation or coating and point at which the pigment is released. The
ranges of encapsulating material and methods suitable for colours are carbohydrates,
gums, phospholipids and proteins and spray drying (SD), spray coating, spinning disc
technology, freeze-drying (FD) respectively.

Curcumin

Curcumin (1-7-bis (4-hydroxy-3-methoxy phenyl) –1,6- heptadiene-3, 5-dione) is

a natural yellow orange dye extracted from the rhizome of plant Curcuma longa L.
(Zingiberaceae). It is a natural compound primarily used for food coloring. Further, the
substance has pharmacological properties such as antitumor and anticancer activities. It is
also a photodynamic agent used for the destruction of bacterial and tumor cells.
Curcumin is non-toxic even at high dosages. Other than food colourant, it is used as
antioxidant additives for lubricants and motor oil and in sunscreen compounds (Hoon
Bong, P, 2000).

Curcumin and curcuminoids are practically insoluble in water at neutral and

medium acid condition. On the other hand, it is soluble in alkali or in glacial acetic acid.
However, its colour is not constant in aqueous medium or in organic solvents due to
degradation or conversion to its dissociation forms (Tonnesen and Karlsen, 1985).
Alkaline degradation of curcumin (CC), demethoxy curcumin (DMC) and Bis-
demethoxy curcumin (BDMC) in aqueous solution follows first order kinetics. This
response is also similar for pigments combined or alone in solution. Degradation rate
constant rapidly increases from pH 7.45 to a maximum at approximately at pH 10.2 and

7
 Chapter One

then tends to decline slightly at higher pH. The rate of alkaline degradation follows
CC>DMC>BDMC in increasing order (Prince and Buescher, 1997)

Prince, 1996 had studied the light induced degradation of CC, DMC, and BDMC
by exposing purified pigments to light of 1450-lux intensity. On analytical separation and
molar quantification on reverse-phase HPLC, they had reported that stability of curcumin
depends on the solvent system and presence of oxygen that in turns follows first order
kinetics. In the same study curcumin solution in methanol sparged with air. The stability
of curcuminoids against photo-oxidation was in the following increasing order
CC>DMC>BDMC. The degradation products were later identified to be a cyclization
product, vanillin, vanillic acid, ferulic acid, ferulic aldehyde and 4- vinylguaiacol.

Thus, use of curcumin in foods as coloring agent, pharmaceuticals and as a drug is

limited due to its low solubility under acid and neutral condition, high decomposition in
alkaline condition, oxidation in presence of oxygen and photodegradation in organic and
aqueous solutions.

8
 Chapter One

Strategies for Stabilization of Curcumin

Few attempts have been made for achieving stability of curcumin. Manabe et al.,
1974, enhanced the stability of curcumin by adding ascorbic acid with it showing
optimum residual ratio and permeation into peeled chestnuts as well as in syrup. Solution
of curcumin in propylene glycol has been reported to be more stable than its powder
counterpart (Sowbhagya et al., 1998)

Curcumin is also stabilized against colour change by maintaining it at pH range of

from about 3.5 to 4.5 which comprises a spray dried intimate mixture of curcumin, an
organic acid, a buffer, a dispersant for curcumin and a film forming encapsulate.
(Richard, 1980).

Curcuminoids are not affected by water activity. It was shown that there was no
influence of water activity on the stability in curcumin and turmeric oleoresin-
microcrystalline cellulose model system. (Souza et al., 1997). A patent involves the
formation of metal-curcumin coloring complexes, which are water soluble and more
stable than curcumin. (Maing and Miller, 1981).

In one study, cyclodextrin complex of curcumin was found to improve the water
solubility, hydrolytic and photochemical stability of compound (Tonnesen et al., 2002).
However, photodecomposition rate increases compared to curcumin solution in organic
solvent as the cavity size as well as charge and bulkiness of cyclodextrin side chain
influence the stability of complexion and degradation rate of curcumin. Therefore, it is of
interest to study the use of other polysaccharides like gum acacia as wall material for
micro-encapsulation of curcumin with additives.

Gum Acacia (GA) is a hydrocolloid obtained as natural exudates of acacia tree.

GA is composed of a highly branch arrangement of simple sugar galactose, arabinose,
rhamnose and glucoronic acid (Aderson and stodard, 1966; Street and Aderson, 1983). It

9
 Chapter One

also contains proteinaceous material covalently attached to the polysaccharide component

with an enhanced ability to absorb at oil-water interface and stabilizing effect on
emulsion. This is due to the hydrophobic nature of proteinaceous chain, which absorbs at
the surface of the oil droplets, while the hydrophilic carbohydrate blocks attached to
protein chain protrude into aqueous solution preventing droplet aggregation and
coalescence (Islam et al., 1997). GA is also good encapsulant due to its high water
solubility, low viscosity of concentrated solution relative to other hydrocolloids gums and
ability to act as oil in water emulsion (Gliksman, 1983).

Literature survey showed that the approach of stabilization of curcumin by use of

GA as encapsulant with antioxidants like TBHQ, Ascorbyl plamitate and their
combination by SD as well as by FD has not been explored. In present study, preparation
of Spray Dried Encapsulated Curcumin (SDEC) and Freeze Dried Encapsulated
Curcumin (FDEC) with and with out additives and study of their stability against
exposure to severe conditions like UV (365nm) and heat (45 0C) has been envisaged. It
was conceptualized that encapsulation with GA could impart physical protection and act
as a barrier to light as well as O2. Also, it was thought that antioxidant additives could
further stabilize the pigment against photo-oxidation.

10
 Chapter Two

CHAPTER 2
MATERIALS AND METHODS

MATERIALS REQUIRED:

Curcumin Natural flavours and essences, Mysore

Gum Acacia Gum acacia-71, Alok Chemicals

Corporation, India

Ethyl Alcohol 95 percent

Tertiary Butyl Hydroxy Quinone (TBHQ) Sigma-Aldrich, USA

Ascorbyl Palmite (AP) Sigma-Aldrich, USA

Distilled Water Double distilled

Refine Sunflower oil

Tween 60 Sigma-Aldrich, USA

EQUIPMENTS USED:

UV Lamp Allen 425 modelLCF-750-Q

UV-VIS Spectrophotometer GBC Cintra 10

Ultra-Sonication Branson Model 1510

10
 Chapter Two

LSD-48 Mini Spray drier JISL

Freeze Drier Lyophilization Model-503, USA

Homogenizer Ultra-Turrax, T-25 Basic, Ira-Werke,

Germany

METHODS:

Product preparation

a) Preparation of Emulsion

Emulsion was prepared at room temperature as described by Mc Nanee et al., 1998,

with some modification. Emulsion dispersed aqueous phase consisted of GA (10%) and
Tween 60. In addition to this oil phase consisted of refined sunflower oil (2.7% w/v),
Curcumin (0.5% w/v) and additives in equal ratio to curcumin, were mixed with aqueous
phase using high shear and subsequently homogenized at 16000 rpm in homogenizer
(Ultra-Turrax T-25 Basic) for 10 minutes. Then emulsion was kept at low temperature in
dark condition until it was subjected to drying.

b) Spray drying of Emulsion

For preparation of micro-encapsulated curcumin, prepared emulsion with or without

additives was fed to LSD-48 Mini Spray drier at rate of 2 ml/ minutes, 2 kg/cm2 with
inlet temperature of 142± 5 °C and outlet temperature at 80± 3 °C. Spray dried product
was collected in sealed plastic bags and kept at dry cool and dark place.

11
 Chapter Two

c) Freeze drying of Emulsion

Emulsion was frozen to -30°C. The freeze drier Lyophilization Model-503, USA
operated at pressure of 250 mtorr. The temperature was increasing stepwise in following
sequence i.e. -30°C for 2 hours the –20°C to 15 °C within 18 hours. Dried sheets of
microencapsulated curcumin were obtained at ambient temperature, collected in sealed
plastic bags, which were subsequently powdered by hand pressing to uniform size
particle size. The product was stored at a dry cool and dark place.

Sample Preparation

Determination of total colour value of Freeze Dried Encapsulated Curcumin

(FDEC) control and FDEC with AP + TBHQ in equal amounts was done in duplicate as
per American Spice Trade Association (ASTA) method with some modification. In that
0.2 g of sample was taken, to which 80 ml of 95% ethanol was added. Subsequently
contents were refluxed for 2½ hours. After refluxing, the contents were cooled to room
temperature and volume was made to 100 ml in volumetric flask that was kept overnight
in dark condition to have a clear supernatant. Sample was taken from it and was diluted
ten times then absorbance (OD) was taken at 425 nm in spectrophotometer (UV-VIS
Spectrophotometer GBC Cintra 10).

In an alternative method, determination of total colour value for all encapsulated

formulation of curcumin with and without additive was done by Sonication method that
was developed for the faster estimation of colour value. A known weight (0.2 g) of FDEC
or SDEC was taken in a 250 ml conical flask and 30 ml of 95% alcohol added, which in
turn was sonicated in water bath at room temperature for 5 and 10 minutes respectively.
Subsequently contents were transferred to a 100 ml volumetric flask with 2-3 washings.
The volume was made up to 100 ml and content is kept for overnight in dark. Then
sample was taken from it that was diluted to 1:10 times and absorbance was taken at
425nm in Spectrophotometer.

12
 Chapter Two

% Curcumin = (Volume) x (Dilution) x (Absorbance at 425nm) x (0.0025x100)

(Weight of sample) x (1000*0.42)

Quantification of % Retention of curcumin on exposure to UV light and Heat

FDEC and SDEC with and without additives were exposed to UV light at 365nm
(UV Lamp Allen 425 modelLCF-750-Q) and 45°C temperature in incubator. Samples
were mixed thoroughly and known weight (0.2 g) was drawn at different time interval
then colour value was determined by ultra-sonication method at different period of
exposure. In addition to this colour value of different formulation of FDEC and SDEC
without exposure (0 hours) was also be determine for calculation for percentage
retention. The whole study was done in duplicates and average value is reported.

% Degradation = (CV of sample without exposure) – (CV of sample with exposure) x100

(CV of sample without exposure)

% Retention = 100 – (% Degradation)

13
 Chapter Three

CHAPTER 3

RESULTS AND DISCUSSION

The yield of microencapsulated curcumin for different formulations for FD and

SD method is shown in table 3.1. From the table it is evident that the yield of FD method
of encapsulation is lesser than SD method of encapsulation. However, practically the
yield of product from freeze-drying is always more than products obtained from spray
drying. The reason for lesser yield in the present case was the losses during the scrapping
of freeze dried material from the tray. Because of the atmospheric moisture, condensation
occurred on the thin bottom layer during the course of scrapping, which became sticky
mass and that resulted in incomplete product recovery.

Table 3.1 CONTROL TBHQ AP AP + TBHQ AVERAGE

YIELD % FD 41.21 36.49 35.03 45.54 35.57

SD 37.20 30.71 43.06 40.89 37.97

Sonication using ultrasound is an efficient non-thermal method for extraction of

compounds from matrices as it creates intense agitation, which causes the total dispersion
of solids and improves the material transfer. Ultrasound was used for disruption of cells
and tissues in migration of proteins, minerals and other substance into solutions (Huges,
D. E. and Nyborg, W. L., 1962, Kim, S. M and Zayas, J. F., 1989). Rozzi et al., 2002 had
used sonication for extraction of lycopene from tomato peel for preparation of standard
solution. Thus, this method is extensively used for extraction of compounds from solid
material or matrices into solvent.

14
 Chapter Three

The comparison between ASTA and sonication method for estimation of colour
value was made for control and formulation with both the additives (Table 3.2). It was
found that there was little difference between the colour values obtained from both the
methods. Therefore, sonication method, which was simpler and consumed lesser time,
was employed further for estimation of curcumin in all formulations.

Table 3.2 Control AP + T

Method I II Average I II Average
ASTA 1.9621 2.088 2.025 2.288 2.580 2.434
Ultra-Sonication 1.984 2.054 2.019 2.344 2.45 2.401

While comparing the stability offered by both FDEC and SDEC control against
UV light at 365nm and 450C incubation, it can be inferred from Table 3.3 and figure1
that stability of FDEC and SDEC against UV light is almost same. However, in case of
stability against thermal exposure SDEC exhibited more stability than FDEC (Table 3.4
and figure 2). This difference in stability of freeze and spray dried products is probably
due to morphological difference that exists in the constitution of both products. In case of
spray dried product, case hardening might have occurred during at high temperature of
drying which would allow lesser penetration of heat within the matrix. However, freeze
dried product that is more porous in nature, offered lesser resistance to heat transfer
within the particles, which probably could be the cause for higher degradation of
curcumin in this product.

In case of UV light exposure it was evident that negligible difference exists

between the stability offered to curcumin by spray as well as freeze dried encapsulation.
This is apparently due to the UV light exposure being more severe than thermal exposure.
In addition to this, UV light can penetrate within the particle of spray and freeze dried
product to the similar extent leading to decomposition of curcumin to the same level.
However, these have to substantiated further by investigations using SEM studies of

15
 Chapter Three

encapsulated curcumin. The stability of encapsulated curcumin against UV light at 365

nm is more than that in solutions, both organic and aqueous systems, reported by
Kumawat, A. and Srinivas P., 2002.

Table 3.3
% RETENTION OF ON EXPOSURE TO UV light @ 365mn
HOURS
OF FREED DRIED
EXPOSUR ENCAPSULATED CURCUMIN FREED DRIED ENCAPSULATED
E (FDECuv) CURCUMIN (SDECuv)
0 100 100
96 96.13 92.96
168 94.7 92.3
240 88.51 87.93
336 86.73 85.48
556 79.31 82.11
724 75.78 75.71

Figure 1:EFFECT OF UV LIGHT EXPOSURE ON

CONTROLS
% RETENTION OF CURCUMIN

97

92

87
FDECuv
82 SDECuv

77

72
0 150 300 450 600 750
HOURS OF EXPOSURE TO UV @365nm

16
 Chapter Three

Table 3.4
% RETENTION OF ON EXPOSURE TO HEAT AT 450C
FREED DRIED SPRAY DRIED
HOURS OF ENCAPSULATED ENCAPSULATED
EXPOSURE CURCUMIN (FDECuv) CURCUMIN (SDECuv)
0 100 100
96 88.5 91.58
240 83.67 86.65
556 80.36 82.91
724 75.95 78.63

Figure 2 : EFFECT OF HEAT EXPOSURE ON CONTROLS

97
% RETENTION OF CURCUMIN

92

87 FDECth
SDECth

82

77

72
0 150 300 450 600 750
HOURS OF EXPOSURE TO 45 C

17
 Chapter Three

From Table 3.5 and figure 3, it can be concluded that stability offered by different
formulations with additives of FDEC and SDEC against UV light at 365nm exposure is
in decreasing order as follows AP+T > AP>TBHQ respectively.

Table 3.5
% RETENTION OF ON EXPOSURE TO UV LIGHT AT 365mn
FREED DRIED SPRAY DRIED
HOURS OF
ENCAPSULATED CURCUMIN ENCAPSULATED CURCUMIN
EXPOSURE
TBHQ AP AP+T TBHQ AP AP+T
0 100 100 100 100 100 100
96 94.74 79.59 98.31 87.4 96.64 96.48
168 94.22 78.95 97.5 85.06 90.44 96
240 93.12 78.6 97.4 84.48 86.18 93.01
336 91.55 78.1 96.62 81.13 84.91 93.33
556 85.03 75.1 96.54 79.29 78.94 90.53
724 73.017 73.622 95.46 71.7 76.14 89.54

F ig u r e 3 : E F F E C T O F U V L IG H T E X P O S U R E
O N F O R M U L A T IO N S

T B H Q fd A p fd A P + T fd
TBHQ sd Apsd AP+Tsd
100
% RETENTION OF CUCUMIN

90

80

70

0 150 300 450 600 750

HO URS O F EXPO SURE TO UV @ 365nm

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 Chapter Three

Stability offered by different formulations of FDEC against 450C exposure is also

in the decreasing order of AP+T > TBHQ > AP. However, for different SDEC
formulations, AP offers more stability than TBHQ (Table 3.6 and figure 4).

Table 3.6
% RETENTION OF ON EXPOSURE TO 450C
FREED DRIED ENCAPSULATED SPRAY DRIED ENCAPSULATED
HOURS OF
CURCUMIN CURCUMIN
EXPOSURE
TBHQ AP AP+T TBHQ AP AP+T

0 100 100 100 100 100 100

96 93.36 87.79 98.11 85.49 97.68 98.04

240 90.95 83.14 97.39 83.93 93.1 95.64

556 85.94 78.22 96.54 81.81 88.21 93.49

724 84.68 76.98 94.24 74.46 79.25 90.71

Figure 4: EFFECT OF HEAT EXPOSURE ON

FORMULATIONS

TBHQfd Apfd AP+Tfd

TBHQsd Apsd AP+Tsd
100
% RETENTION OF CURCUMIN

90

80

70
0 150 300 450 600 750

HOURS OF EXPOSURE AT 45 C

19
 Chapter Three

Thus, by comparing the data of stability of encapsulated curcumin with and

without additives it can be inferred that stability of curcumin in matrix is further
enhanced by the presence of different additives, TBHQ (T), Ascorbyl palmitate (AP) and
their combination. In addition to this, the combination of TBHQ and Ascorbyl palmitate
is more effective than either of additive when used alone.

In a similar study by Kumawat, A and Srinivas, P., 2002, it was shown that use of
antioxidant stabilized the curcumin in acetone system exposed to UV light at 365 nm and
their NMR spectral studies had indicated the association of protons of the colorant with
that of the additive and indication of photo-catalyzed transformations. It was concluded
that color instability arises from the free access of the radicals to the chromophore and
related transformation reactions and presence of additives, such a process is inhibited,
which lends better stability to the colorant.

Present experimental results also indicate that the color degradation by light and
heat is probably a radical mediated reaction, which causes the formation of products that
alter the structure of the chromophore and hence affect the color of the colorant. The
mechanism of degradation of curcumin under the influence of heat and UV light, also its
stabilization by additives is to be studied in depth by spectroscopic studies like NMR and
MS for a better understanding of the chemical processes involved both in degradation of
the pigment and its stabilization by anti-oxidants.

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 Chapter 4

SUMMARY AND CONCLUSIONS

For the use of natural colors in food products, it is important to have a complete
understanding of the chemical and physical environment that exist in the product(s) to be
colored during and after processing. Also the capabilities and limitations of natural colors
as they apply to the product(s) need to taken into consideration. Instability of natural
colors is one of the major limitations in the application of these colors.

A good number and variety of natural color pigments are now available for
coloring foods. The global market for natural colors is estimated at more than $939
million. Despite processing limitations and higher costs, the natural food colorant market
is growing and this growth is largely influenced by the consumer perception that ‘natural
is superior’.

Curcumin colour pigment is highly unstable to light and less stable to heat. In the
present study, effect of encapsulation of curcumin along with different additives viz.,
TBHQ, Ascorbyl palmitate and their combination on the color stability of curcumin has
been investigated.

Following conclusions can be drawn from this study:

Encapsulation of curcumin offers stability against exposure to UV light and

heat

Stability of curcumin is further enhanced by the presence of different additives,

TBHQ (T), ascorbyl palmitate (AP) and their combination.

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 Chapter 4

Combination of AP and T is much more effective than either of the additives

showing synergistic action.

This methodology for stabilization of curcumin pigment is novel

Results indicate that free radical mediated oxidative reactions occur during
photo and thermal degradation

The mechanism of degradation of curcumin and its stabilization by additives

has to be studied in depth by NMR Spectral studies

Further studies are necessary for arriving at the optimum concentration,

economics and regulatory levels for these additives.

22
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