COLOURANTS

Introductions
Pigments in Animal and Plant
Tissues
Effects of processing
Color
Color and appearance are major, even
sometimes the most important, quality
attributes of foods
First to be evaluated by customers
Customers often relate specific colors of
food to quality, e.g. maturity of fruits,
freshness of raw meat
Influences flavor perception, e.g. red ≈
berry flavor, yellow ≈ orange flavor, etc
Colourants
Colorant: any chemical, either
natural or synthetic, that gives color
Foods have color because the are able to
reflect or emit different quantities of energy
at certain range of wavelengths stimulating
the retina in the eye.
Wavelengths of visible light: 380 – 770 nm
Some colorants, e.g. β-carotene or
riboflavin, are also nutrients
Food Color
• Any substance that is added to food
to change or enhance its color.
1) Natural colors
2) Synthetic colors
3) Lakes and dyes
• Natural Food color is any dye,
pigment or any other substance
obtained from seeds, fruits,
vegetables, algae or insect
Natural pigments; carotenoids,
anthocyanins, chlorophylls,
polyphenols, turmeric, caramel.
Pigments, Dyes & Lakes
Pigment: naturally occurring colourant found in the
cells of plants and animals
In food industry: dye = a certified food grade
water-soluble colorant
Lakes are oil-dispersible dyes extended on a
substratum
The only approved substratum for FD&C lakes:
alumina
Uncertified colorants may also be used given that
they are natural pigments or synthesized
substances, identical to the natural pigment
Stability of Natural Pigments
Many are unstable during processing and storage
Prevention is usually difficult, or even impossible
Depending on the pigment, stability is affected by:
Light
Oxygen
Heavy metals
Oxidizing or reducing agents
Temperature
Water activity
pH
Irradiation
Primary effects: formation of free radicals and excited molecules
Changes depend on many factors
The behavior of individual food colorants in solutions or extracts toward
irradiation may not predict the color changes
More sensitive: carotene
Pressure
Effects on foods: destruction of microorganisms, denaturation of proteins,
alteration of enzymatic reactions
More sensitive: myoglobin
Different stability performed by different pigment under different
conditions
Instability of natural pigments causes colorants are often added to
processed foods
Classification of natural colorants
Group Classification

Isoprenoid Carotenoids, Xanthophylls

derivatives
Tetrapyrrole Chlorophylls, Porphyrin, Heme pigments,
derivatives Bilins
Benzopyran Anthocyanins, Flavones, Flavonoids,
derivatives Tannins
Artefacts Melanoidins, Caramels

Others Anthraquinone, Betalain, Phenalone, etc

® CRC Press, Colour in Food, 2002

 Pigments

Isoprenoid Derivatives
Isoprenoid Derivatives

Multiples of (usually) eight isoprene units

(C5H8)
A chain of two terpenoids, each terpenoid
(a 10 carbon unit) is formed from 2
isoprene units being joined head to tail
Isoprenoid Derivatives – Carotenoids

Carotenoids: tetraterpenoids
Most widespread pigments
Functions in photosynthesis and photoprotection (in plan
tissues) and as precursors of vitamin A (nutrition)
Classification:
Hydrocarbons: carotenes & lycopene  tend to be fat soluble
and not soluble in water
Oxygenated: xanthophylls  Contain hydroxyl, epoxy, aldehyde
and keto groups
The structure of xanthophylls is essentially the same as the
carotenes but they contain hydroxyl groups  thus, they are
polar and more soluble in ethanol than the carotenes
Isoprenoid Derivatives – Carotenoids –
cont’d
Most common in plants: β-
carotene (also used as
additive colorant)
Distribution: tomatoes
(lycopene), carrots (α- and β-
carotenes), red peppers
(capxanthin), corn (lutein and
zeaxanthin), etc
Concentration correlates with
chlorophyll content
Factors affecting
concentration: maturity,
exposure to light, climate,
pesticide and fertilizes use,
soil type
Isoprenoid Derivatives – Carotenoids –
cont’d
Properties:
Soluble in oils and organic compounds
Have antioxidant properties
Moderately heat stable
Color loss by oxidation
Similar to unsaturated fatty acids, carotenoids can
undergo oxidation via a radical reaction mechanism
 formation of epoxide and isomeration of epoxide to
a furanoxide
E.g. loss of color in canned fruit juice
Can undergo Cis/Trans isomeration by heat,
acid, and light
Relatively stable during storage and handling
 Carotenoids Stability
The presence of multiple unsaturated
carbon–carbon double bonds makes
carotenoids susceptible to oxidation catalysed
by light, metals and hydroperoxides.
Oxidation results in the bleaching of colour,
loss of vitamin A activity and off odours.
Stable up to 50°C and in the pH range of 2–7,
thus, not degraded by most forms of
processing.
With heating, the naturally occurring trans
isomer rearranges to the cis isomer.
 Pigment

Benzopyran and Tetrapyrrole

Derivatives
Benzopyran Derivatives
Sub-structure:
anthocyanidins,
flavonoids, tannins
Basic common
structure: ring
structure
Anthocyanins:
Ring + some hydroxyl
groups through which
are conjugated sugar
molecules
Refers to the molecules
without the glycoside
residue
Benzopyran Derivatives – cont’d
Flavonoids: contain
a keto or hydroxyl
group at position 4
Tannins:
Found in tree bark
and leaves of many
plants
Responsible for the
formation of the
black/dark color of
tea and the brown
color of old wines
Benzopyran Derivatives – cont’d
Benzopyran Derivatives –
Anthocyanins
Responsible for a range
of colors in plants: blue,
purple, violet, magenta,
red, orange
The sugar moiety can
be hydrolyzed 
anthocyanidin
Anthocyanins are
soluble in water and
alcohol, insoluble in a
polar solvent and
unstable in neutral or
alkaline medium.
Benzopyran Derivatives –
Anthocyanins – cont’d
Benzopyran Derivatives –
Anthocyanins – cont’d
STABILITY
Relatively unstable
Most stable in acidic conditions
Degradation factors:
Major: pH, temperature, oxygen
Minor: light (accelerates degradation), enzymes
(discoloration), ascorbic acid, sulfur dioxide
(bleaching effect), metal ions (complexation
with metals stabilizes or discolorizes color),
sugars (stabilizer)
Oxygen
Unsaturated  susceptible to oxidative
degradation during various steps of processing
and storage
Factors affecting pigment loss
Factors should include the effects of oxidation, temperature
change, pH change and the presence of metal ions.

Anthocyanins:
They are most stable at low pH and temperature.
Anthocyanin pigments readily degrade during thermal
processing which can have a dramatic impact on colour
quality and may also affect nutritional properties.
Anthocyanins are enzymatically degraded in the presence of
polyphenol oxidase. This enzyme can be inactivated by mild
heating and therefore the inclusion of a blanching step
(heating to approximately 50C) can have a positive effect on
anthocyanin retention.
>50% of anthocyanins were lost after 6 month of storage
due to extensively polymerized during storage.
Benzopyran Derivatives –
Anthocyanins – cont’d
pH changes:
The most affected group  due to
the ionisable groups (groups with
charged atoms) in anthocyanins
Effects of pH depends on the type
of anthocyanin, generally:
In low pH, the color of anthocyanins
is red
As the pH increase, anthocyanins
may undergo two possible
pathways:
– Deprotonation  resulting in a blue
quinoidal compound
– Hydration  resulting in the
colorless chalcone
Benzopyran Derivatives –
Flavonoids
Flavonoids are the yellow plant pigments seen most
notably in lemons, oranges, and grapefruit.
The name stems from the Latin word "flavus," which
means yellow.
Flavonoids are placed into 12 different classes, the
best known of which are the anthocyanins, flavonols,
and flavones. All flavonoids have 15 carbon atoms
and consist of two 6-carbon rings connected to one
another by a carbon ring which contains an oxygen
atom.
Most naturally occurring flavonoids are bound to one
or more sugar molecules. Small changes in a
flavonoid's structure can cause large changes in its
color.
Proanthocyanidin – beige color (bean seed coat),
shades of black, red, brown and tan (cocoa,
cinnamon, grape seed and skin)
Benzopyran Derivatives –Tannins

Water-soluble polyphenolic compounds

Able to combine with proteins and other
polymers, e.g. polysaccharides
Color range: yellowish-white to light brown
Contribute to astringency in foods
Special role in formation of black color in
tea due to the conversion of catechin (a
tannin) to aflavins and arubigins
Tetrapyrrole Derivatives
Common structure: a central metal atom
surrounded by four linked pyrrole rings
(tetrapyrrole)  a porphyrin ring
2 important groups:
The chlorophylls: the central metal is
magnesium
The hemicromes (hemoglobin and myoglobin):
central metal is iron
the porphyrin ring forms the prosthetic group attached
to the globular protein globin
Tetrapyrrole Derivatives –
Chlorophylls
Characteristics
Major pigments in green
plants, algae and
photosynthetic bacteria
Located in chloroplasts
Weakly linked to
carotenoids, lipids and
lipoproteins
Best extracted with polar
solvents
Susceptible to
photodegradation once the
protection of surrounding is
gone
Chlorophylls Stability
Chlorophylls are susceptible to many chemical or enzymatic
degradation reactions.
The simultaneous actions of enzymes, weak acids, oxygen,
light and heat can lead to the formation of a large number of
degradation products.
Major chemical degradation routes are associated with
pheophytinization, epimerization, and pyrolysis, and also with
hydroxylation, oxidation or photo-oxidation, if light is
implicated
the main cause of green vegetable discolouration during
processing is the conversion of chlorophylls to pheophytins
when heated and/or by the infuence of pH (acidic condition)
Chlorophyll a was more susceptible to thermal degradation
than chlorophyll b in acidic conditions
Tetrapyrrole Derivatives –
Chlorophylls – cont’d
Degradation - Enzymatic changes
By the enzyme chlorophyllase, that cleaves phytol
The cleavage of the phytol chain of chlorophyll by
the enzyme chlorophyllase results in the formation
of chlorophyllide
In the presence of acid, chlorophyllides undergo
loss of the magnesium and form phephorbides
It is reported that this enzyme acts at ambient
temperature only in the presence of high
concentrations of organic solvents and only at
temperatures between 65° and 75 °C in aqueus
medium.
On the other hand, chlorophyllase losses its
activity when heated to 100 °C
Tetrapyrrole Derivatives –
Chlorophylls – cont’d
Degradation - by heat and acid
2 groups of derivatives formed during thermal
processing:
Mg-containing: green
Mg-free (pheophytin): olive-brown
Factors: media, pH, temperature range
Further heating produces pyropheophytin
In vegetable tissue, degradation is affected by pH.
Chlorophyll is very stable in basic media (pH 9.0)
Often happen in thermal processing, e.g.
blanching and sterilization (the degree of loss can
be as much as 80-100%)
• When green vegetables are cooked and/or
exposed to acid, the magnesium gets removed
from the center of this ring structure and replaced
by an atom of hydrogen.
• This results in a change from chlorophyll to
pheophytin.
• Chlorophyll a gets turned into a molecule called
pheophytin a, and the chlorophyll b gets turned
into pheophytin b.
• The color of the vegetable changes from bright
green to olive-gray. (The pheophytin provides a
green-gray color, and the pheophytin b provides
an olive-green color).
Tetrapyrrole Derivatives –
Chlorophylls – cont’d
Color Preservation in Food Processing
Acidification
HTST
Enzymatic conversion of chlorophyll
to chlorophyllide
Application of metallo complexes 
not approved world-wide
Regreening of thermal-processed
vegetables
Tetrapyrrole Derivatives - Heme
Compounds
The color of meat
Types: myoglobin &
hemoglobin
Most hemoglobin is
removed when animals are
slaughtered and bled 
myoglobin becomes the
primary pigment (90%) of
meat
Variation of myoglobin
quantity is affected by:
Species,
Age,
Sex, and
Physical activity of the
animals
Myoglobin binds oxygen more tightly than
hemoglobin. It is not good for oxygen transport
and is instead good for oxygen storage.
Hemoglobin is the opposite- good for 02 transport
but not for storage. This is because hemoglobin
has a different affinity for oxygen and therefore it is
released easily.
The myoglobin and hemoglobin have different
structural makeup (this explains why they have
different functions). Myoglobin is also found in
muscle tissue whereas hemoglobin is found in red
blood cells.
Myoglobin 
• Is a monomeric protein and binds molecular
oxygen and carry to muscle tissues.
• Muscle cells use myoglobin to exchange oxygen
during active respiration.

Hemoglobin 
• Is a tetrameric protein and binds molecular
oxygen on RBCs. Being a tetramer it binds four
oxygen molecules and distribute them throughout
the whole body.
• It serves to deliver oxygen needed for cellular
metabolism and removes the resulting waste
product, carbon dioxide from the body tissues. 
Each hemoglobin protein is made up
subunits called hemes, which are
what give blood its red color.
More specifically, the hemes can bind
iron molecules, and these iron
molecules bind oxygen.
The blood cells are red because of
the interaction between iron and
oxygen.
Tetrapyrrole Derivatives - Heme
Compounds – cont’d
Chemistry & Color - Oxidation
Meat color is determined by:
The chemistry of myoglobin
Myoglobin’s state of oxidation
Type of ligands bound to myoglobin’s heme
The state of the globin protein
2 forms of heme irons: reduced ferrous (Fe2+) or
oxidized ferric (Fe3+)
O2 + myoglobin  oxymyoglobin (MbO2) =
oxygenation (not oxidation!!)
In heme compounds, the central iron atom may be
oxidized relatively easily to form metmyoglobin (from
Fe2+ to Fe3+)
Oxidation: Conversion of Fe2+ to become Fe3+
Tetrapyrrole Derivatives - Heme
Compounds – cont’d
Chemistry & Color - Oxidation
Factors affecting oxidation rate:
Presence of oxygen
Presence of globin protein
pH value
Ratio between Mb and MbO2
Presence of trace metals, esp. Cu ions
Tetrapyrrole Derivatives - Heme
Compounds – cont’d
Cured Meat
End result: stable pink color
Reactions:
Nitric oxide (NO) + Mb  nitric oxide myoglobin
(MbNO2) (also known as nitrosylmyoglobin) …
bright red & unstable
Heating: formation of nitric oxide
myohemochromogen (nitrosylhemochrome) …
pink & stable
Too much nitrous acid: formation of
nitrimyoglobin (NMb)  heating 
nitrihemin …green (“nitrite burn”)
Tetrapyrrole Derivatives - Heme
Compounds – cont’d
Tetrapyrrole Derivatives - Heme
Compounds – cont’d
Stability of heme compounds
Factors
Interaction between factors in meat
Environment
Light
Temperature
– Denaturation of protein
– Complete denaturation is at around 80 – 85 0C
– May contain Fe2+ (similar color to oxymyoglobin) or Fe3+
(brown)
Humidity
pH
– pH changes affects the globin protein
Bacteria
Lipid oxidation
Antioxidants
Biochemical factors
MA packaging helps in stabilizing meat color
Natural Colourants

Melanins, Melanoidins and

Caramels
Melanins, Melanoidins and Caramels
In general: Complex polymeric molecules
Melanins
Occur naturally
Responsible for many of the black, grey and brown colors of plants and
animals
Melanoidins and caramels: formed by non-enzymatic browning
reactions, usually during the heat processing of foods
Melanoidins: the result of Maillard reactions (a reaction between a
reducing sugar and a primary or secondary amine)
Caramels: the result of caramelization (a reaction result from the
effect of heat on sugars
Natural Colorants

Others
Betalains
Pigments found in
beetroots
Subdivision:
Betacyanins: purplish red
Betaxanthins: yellow  do
not have an aromatic ring
system attached to N-1 or
sugar residues
Water soluble
The color is not affected by
pH
Heating causes
isomeration to isobetanin
 loss of color of beets
during thermal processing
Oxygen accelerates color
loss
Anthraquinone (Quinones)
Quinones: phenolic
compounds widely
distributed in
plants, esp. trees
(give the color of
wood)
Bitter taste
E.g. cochineal or
carmine
Xanthones
Yellow, phenolic
pigment
Often confused
with quinones and
flavones
E.g. mangiferin in
mangoes,
mangostin in
mangosteens