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Antioxidants in Bakery Products: A Review

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Antioxidants in Bakery Products: A Review

B. Nanditha a; P. Prabhasankar a
a
Flour Milling, Baking, and Confectionery Technology Department, Central Food Technological Research
Institute, Mysore, India

Online Publication Date: 01 January 2009

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 Critical Reviews in Food Science and Nutrition, 49:1–27 (2009)
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ISSN: 1040-8398
DOI: 10.1080/10408390701764104

Antioxidants in Bakery Products:

A Review

B. NANDITHA and P. PRABHASANKAR

Flour Milling, Baking, and Confectionery Technology Department, Central Food Technological Research Institute, Mysore
570020, India
Downloaded By: [CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE] At: 04:07 24 October 2008

Fats impart taste and texture to the product but it is susceptible to oxidation leading to the development of rancidity and
off-flavor. Since ancient times it has been in practice to use antioxidants in foods. Discovery of synthetic antioxidants has
revolutionized the use of antioxidants in food. The effect of these antioxidants in bakery products were reviewed and found
to be effective in enhancing the shelf life. Animal experimental studies have shown that some of the synthetic antioxidants
had toxigenic, mutagenic, and carcinogenic effects. Hence there is an increasing demand for the use of natural antioxidants
in foods, especially in bakery products. Some of the natural antioxidants such as α-tocopherol, β-carotene, and ascorbic
acid were already used in bakery products. These natural antioxidants are found to be effective in enhancing the shelf life of
bakery products but not to the extent of synthetic antioxidants. Baking processing steps may lower the antioxidative activity
but techniques such as encapsulation of antioxidants can retain their activity. Antioxidative activity of the plant extracts such
as garcinia, curcumin, vanillins, and mint were reviewed but studies on their role in bakery products were limited or very
few. Hence there is a wide scope for study under this direction in depth.

Keywords antioxidants, bakery, lipid oxidation, vitamins, food processing

INTRODUCTION BHA (Butylated Hydroxy Anisole), BHT (Butylated Hydroxy

Food is an essential commodity for mankind to survive. In
earlier days, it for future use was not in practice but as time ad-
vanced it became part of day-to-day life. During preservation,
a lot of problems were faced especially shelf life. The shelf life
of food is very limited and subjected to varieties of spoilage due
to its composition viz. moisture, lipid, protein, carbohydrates,
etc. As lipids are one of the major components of food, autoox-
idation of lipids and the generation of free radicals are natural
phenomena in biological and food systems.
Historically, the usage of gum guaiac way back in the 1930s
was the first antioxidant approved for the stabilization of ani-
mal fats especially lard (Joyner and McIntyre, 1938; Madhavi
et al., 1996.) Antioxidants have become an indispensable group
of food additives mainly because of their unique properties of
enhancing the shelf life of food products without any damage
to sensory or nutritional qualities. They includes tocopherols
and derived compounds in baked, fried products and vegetable
oils. ß-carotene in butter, butterfat, coconut oil, and corn oil.
Address correspondence to P. Prabhasankar, Flour Milling, Baking, and
Confectionery Technology Department, Central Food Technological Research
Institute, Mysore 570020, India, Phone: 91-821-2517730, Fax:91-821-2517233.
E-mail: prabhasankar@lycos.com
1
toluene), in bakery products and candies. Tertiary-butyl hydro-
quinone (TBHQ), in oils, fats, and meat products. Gallates in
frying fats and oils. Rosmarinic acid, in the form of the herb
rosemary and Italian seasoning mixtures in naturally or mini-
mally processed foods. In recent years, highly advanced tech-
niques of chemical synthesis have made it possible to develop
new synthetic antioxidants such as Trolox-C, a polymeric deriva-
tive of α-tocopherol and anoxomer. The chemical synthesis of
natural products like ascorbic acid and α-tocopherol on a com-
mercial scale have been successful. The synthetic approaches
have been targeted for developing nontoxic, nonabsorbable com-
pounds and incorporating in a wide range of food products. The
increasing use of antioxidants in food preservation and in dis-
ease prevention calls for clear definition of the term antioxidant
(Madhavi et al., 1996).
According to the US Food and Drug Administration (FDA),
antioxidants are defined as “substances used to preserve food by
retarding deterioration, rancidity, or discoloration due to oxida-
tion.” Antioxidant also was defined with a broader perspective as
“any substances when present at low concentration compared to
those of an oxidizable substrate, significantly delays or prevents
oxidation of the substrate” (Halliwell, 1991). The term “oxidiz-
able substrate” includes components in foods and in biological
systems viz., proteins, lipids, carbohydrates and genetic con-
stituents (Halliwell et al., 1995). According to the Prevention of
 2 B. NANDITHA AND P. PRABHASANKAR
Food Adulteration (1976) Act antioxidant means “a substance
which when added to food, retards or prevents oxidative deteri-
oration of food and does not include sugar, cereal, oils, flours,
herbs, and spices” (Khanna, 1987).
People were using bread as staple food from ancient times.
The bread was brought to India around the end of the seventeenth
century and also during the era of British rule when Europeans
started coming and settling in the country for trade. Due to the
industrial revolution, there was an increase in consumption of
bread and allied products and as a result, breadmaking got slowly
organized (Halliwell et al., 1995). The bakery industry in India
is the largest of the food industries with an annual turnover of
about Rs. 3000 crores. Bread and biscuits form the major baked
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foods accounting to over 80% (bread: 50%, biscuits: 30%) of
the total bakery products produced in the country. The quantities
of bread biscuits produced are more or less the same; however,
the value of biscuits is 112 times more than bread. Over 70%
of the total bakery products come from the unorganized sector.
Though the bakery industry in India has been nonexistent for
a long time, the real fillip came only in the latter part of the
twentieth century. Bakery products still remain the cheapest of
the processed ready-to-eat products in the country. The produc-
tion of bread and biscuits has increased from 5.19 lakh tons in
1975 to 18.95 lakh tons in 1990 recording a four-fold increase
in 15 years. Over 84% of the bread and 79% of the biscuits are
produced the unorganized the sector, which is 2–3 times more
than that produced by the organized sector. The growth rate for
bakery products is estimated at an average of 9.8% per annum
(Shukla et al., 2000). The annual production (2005–2006) of
bakery products in India (bread, biscuits, pastries, cakes, buns,
rusks etc.) is 30 lakh tons and that of bread and biscuits is 15
lakh tons and 11 lakh tons respectively (International FoodTec
India).
Bakery and confectionery products are produced from the
dough prepared with flours of cereals and legumes where starch
is the basic major component. Product formulations are by tradi-
tional cooking methods (oven, frying) or non-traditional meth-
ods (microwave, infrared, etc.) (Barbiroli and Mazzaracchio,
1994). A wide variety of bakery products can be found on su-
permarket shelves, such as breads, biscuits, unsweetened rolls
and buns, dough nuts, meat pies, dessert pies, pizza, quiche,
crackers, cookies, and other products (Smith et al., 2004).
In olden days, the baker could adjust the timing of fermenta-
tion and mixing, as well as of proofing and baking conditions, to
adjust for variation of flour properties, yeast activity, ingredients
quality, and room temperature. These adjustments are necessary
to obtain the desired quality of finished baked foods. In today’s
commercial and competitive bakery and mechanization such a
high degree of flexibility is not available or it is available only
at an unacceptable cost. To help stabilize and bring uniformity
of the dough properties that translate into various bakery prod-
ucts, the baker uses various kinds of functional additives. The
industrialization of the food industry, including baked goods,
is the result of the consumer’s demand for products with high
quality, convenience, longer shelf life, easier storage condition,
Table 1 Permitted flour and bread antioxidants in India∗
Max. Limit
Antioxidants for bakery products (PFA/BIS)
Tertiary Butyl Hydroxy quinone (TBHQ) 0.02%
Ascorbyl palmitate 0.02%
Ethyl gallate, propyl gallate, Octyl gallate, 0.01%
or mixture thereof
Butylated hydroxy toluene (BHT) 0.02% (on fat)
Butylated hydroxy anisole (BHA) 0.02% (on fat)
Citric, gallic and tartaric acid 0.01%
Resin guaiac 0.05%
Ascorbic acid (Improvers) 200 ppm
Sorbic acid and its salt (Rope and mold inhibitors) 1.0 g/kg flour
∗ Reference: (Khanna, 1987; Srivastava and Rao, 1997)
and high appeal to sight, touch, taste, and smell. To meet the
above demands the baking industries are required to use func-
tional food additives (Srivastava and Haridas Rao, 1997). Ta-
ble 1 gives permitted flour and bread additives/antioxidants in
India.
Plant and animal foods, in addition to supplying essential
nutrients for the survival of mankind, also possess a variety of
bioactive substances, which are of considerable use from the
standpoint of food science and biology. In food systems, natu-
rally occurring antioxidants impart protection to food compo-
nents against oxidation. The level of antioxidant is critical in
preserving fats containing foods to minimize the development
of objectionable odors and flavors due to rancidity and to pre-
vent or reduce the formation of decomposed products that could
be toxic (Sims and Fioriti, 1977). The development of rancidity
involves the degradation of vitamins (A, C, D, E, and folate), es-
sential fatty acids (linoleic acid), and essential amino acids like
methionine, cystine, histidine, lysine, and tryptophan resulted in
the loss of nutritional value, which can be prevented by the use
of antioxidants (Roubal and Tappel, 1966; Nielsen et al., 1985).
The usage of antioxidants prevents bleaching of labile pigments
such as anthocyanins, carotenoids, myoglobin, chlorophyll, or
browning reaction products in baked foods (Simpson, 1985).
Antioxidants have antimicrobial activities in addition to their
antioxidative properties in baked products. This phenomenon
has received much attention in food microbiology recently. Most
of the antimicrobial effect of antioxidants was focused on stud-
ies using BHA and BHT. Gailani et al. (1980) tested addi-
tional antioxidants on the growth of selected bacterial. Twenty
four species of bacteria (16 gram positive and 8 gram nega-
tive species) were tested against propyl gallate, TBHQ, BHA,
and BHT in solid and liquid systems at concentrations from 50
to 300 ppm. Most of the test organisms were not inhibited by
these antioxidants at 100 ppm but were inhibited by 300 ppm.
These compounds exhibited a broad spectrum of inhibition since
both gram positive and gram-negative organisms were equally
affected. Antioxidants were heat stable (121◦ C) and had bacte-
ricidal activity rather than bacteriostatic activities (Gailani et al.,
1980).
Antioxidants play a major role in enhancing the shelf life,
and preserving the nutritional and organoleptic qualities of
 ANTIOXIDANTS IN BAKERY PRODUCTS
bakery products. Further, more details on antioxidant mecha-
nism, classification, usage, and stability in bakery products are
discussed in this review.
MECHANISM OF LIPID OXIDATION
Autooxidation or lipid peroxidation is a process in which
susceptible lipids are attacked by oxygen leading to complex
chemical changes, resulting in the rancidity and generation of
off-flavors in the food. The basic mechanism of oxidative rancid-
ity or lipid autooxidation has been well established. Atmospheric
oxygen can react spontaneously with organic compounds and
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cause structural degradation, which is ultimately responsible for
the loss in quality in several chemical products of economic or
industrial importance. In baking systems, the spontaneous ox-
idative reaction can result in the deterioration of lipids because
natural fats, oils, and vitamins especially vitamins A, B, C, D,
E, and K, essential oils, cosmetics, and waxes contain a varying
degree of unstauration in their hydrocarbon chains and these un-
saturated sites are susceptible to oxidation. The direct reaction
of lipid molecule with a molecule of oxygen, is a free-radical
chain reaction and the mechanism of autooxidation can be dis-
tinguished in three distinguishing steps: Initiation, Propagation,
and Termination.
Initiation
The autooxidation of a fat is thought to be initiated with the
formation of free radicals if an unsaturated lipid is in contact
with oxygen (Eq. 1). Initiation reactions take place either by the
abstraction of hydrogen radical from allylic methylene group
of an unsaturated fatty acid or by the addition of a radical to a
double bond as shown below.
RH → R• + H• (1)
ROOH → RO• + HO• (2)
2ROOH → RO• + ROO• + H2 O (3)
The formation of lipid radical R• is usually mediated by trace
metals, irradiation, light, or heat. The hydroperoxides undergo
hemolytic cleavage to form alkoxy radicals (RO• ) or undergo
bimolecular decomposition.
Propagation
In propagation reactions, free radicals are converted into other
radicals. The initial formation of one radical becomes respon-
sible for the subsequent chemical transformation of innumer-
able molecules due to chain of events. In fact, propagation of
free-radical oxidation processes occurs in the case of lipids by
3
chain reactions that consume oxygen and yield new free-radical
species (peroxy radicals, ROO• ) or by the formation of perox-
ides (ROOH) as in Equations (4) and (5).
R• + O2 → ROO• (4)
ROO• + RH → ROOH + R• (5)
The products R• and ROO• can further propagate free-radical
reactions. Lipid peroxy radicals (ROO• ) initiate a chain reac-
tion with other molecules, resulting in the formation of lipid
hydroperoxides and lipid free radicals. This reaction, when re-
peated many times, produces an accumulation of hydroperox-
ides.
Lipid hydroperoxides, the primary products of autooxidation,
are odorless and tasteless. Since lipid radicals are also highly
reactive, they can easily undergo propagation reactions by two
mechanisms: by reaction with an oxygen molecule in the triplet
ground state (Eq. 4) or by removal of a hydrogen atom (Eq. 5).
Molecular oxygen is particularly susceptible to radical attack.
This radical-oxygen reaction is very fast, as it requires almost
zero activation energy.
Termination
A free radical has been defined as a molecular entity possess-
ing an unpaired electron. Free radicals are electrically neutral
and solvation effects are generally very small. They are con-
sidered to be bonding-deficient and hence structurally unstable.
Radicals therefore tend to react whenever possible to restore
normal bonding. That is why a free radical is highly reactive
and radicals bond to one another, forming a stable nonradi-
cal compound. Thus, the termination reactions lead to interrup-
tion of the repeating sequence of propagating steps of the chain
reaction.
R• + R• → R − R (6)
R• + ROO• → ROOR (7)
ROO• + ROO• → ROOR + O2 (8)
When the coupling of two radicals involves identical species,
the reaction is recognized as dimerization.
Classification and Mechanism of Antioxidants
Food antioxidants are classified based on their function
(Madhavi et al., 1996) (Fig. 1) as follows.
 4 B. NANDITHA AND P. PRABHASANKAR

Food Antioxidants

Phenols Gallates
Hydroquinone
Primary Trihydroxy butyrophenone
Antioxidants Nordihydroguairetic acid

Secondary/Synergistic
Antioxidants
Hindered Butylated hydroxy anisole
Phenols Butylated hydroxy toluene
Tert iary butyl hydroxy quinone
Toc opherols
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Gum Guaiac A
Ionox series

Miscellaneous Ethoxyquin
Primary Antioxidants Noxomer
Trolox-C

Oxygen Scavengers Chelating Agents Secondary Miscellaneous

Antioxidants Antioxidants

Sulfites Polyphosphates Thiodipropionic Nitrites

Ascorbic acid Ethylene diamine acid Amino acid
Ascorbyl tetra acetic acid Dilauryl, Spice
palmitate Tartaric acid Distearyl esters extracts
Erythrobic acid Citric acid Flavonoids
Citrate esters Vitamin A
Phytic acid β-Carotene
Lecithin Tea extracts
Zinc

Figure 1 Classification of food antioxidants.

Primary Antioxidants AH + R• → A• + RH
AH + ROO• → A• + ROOH
Primary antioxidants terminate the free-radical chain reaction
by donating hydrogen or electrons to free radicals and converting AH + RO• → A• + ROH
them to more stable products. They may also function by addi-
tion in reactions with the lipid radicals, forming lipid-antioxidant The antioxidant free radical may further interfere with the
complexes. Primary antioxidants are effective at very low chain-propagation reactions by forming peroxy antioxidant
concentrations, and at higher levels they may become pro- compounds.
oxidants.
Primary antioxidants may either delay or inhibit the initiation A• + ROO• → ROOH
step by reacting with a fat free radical or inhibit the propagation
step by reacting with the peroxy or alkoxy radicals. A• + RO• → ROA
 ANTIOXIDANTS IN BAKERY PRODUCTS
Hindered phenolic antioxidants have substituted alkyl or
electron-releasing groups in the ortho or para positions in the
aromatic ring, which decreases the reactivity of the –OH group.
The electron-donating groups increase the electron density on
the –OH group by an inductive effect and increase the reac-
tivity with the lipid radicals. Substitution with butyl or ethyl
groups at the paraposition enhances the activity compared to
that of methyl groups. Substitution also reduces the number of
propagation reactions that may occur involving antioxidant free
radicals:
AOO• + RH• → AOOH + R•
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A• + RH• → AH + R•
A• + O•2 → AOO•
Synergistic Antioxidants
Synergistic antioxidants can be broadly classified as oxygen
scavengers and chelators. Synergists function by various mecha-
nisms. They may act as hydrogen donors to the phenoxy radical,
thereby regenerating the primary antioxidant. Hence phenolic
antioxidants can be used at lower levels if a synergist is in-
corporated simultaneously in the food product. Synergists also
provide an acidic medium that improves the stability of primary
antioxidants.
Secondary Antioxidants
Secondary or preventive antioxidants function by decompos-
ing the lipid peroxides into stable end products.
Miscellaneous Antioxidants
Compounds such as flavonoids and related compounds and
amino acids function as both primary antioxidants and syner-
gists. Nitrites and nitrates, function as antioxidants by convert-
ing heme proteins to inactive nitric oxide forms and by chelating
the metal ions, especially nonheme iron, copper, and cobalt. β-
Carotene and related carotenoids are effective quenchers of sin-
glet oxygen and also prevent the formation of hydroperoxides.
Zinc strongly inhibits lipid peroxidation at the membrane level,
by altering or preventing iron binding. Selenium is necessary for
the synthesis and activity of glutathione peroxidase, a primary
antioxidant enzyme.
Based on the sources food antioxidants can be classified as
Synthetic antioxidants, Antioxidants from food packaging ma-
terials, and Natural antioxidants.
Biological Lipid Peroxidation
While antioxidants are of interest to the baking industry, be-
cause they prevent rancidity (Smith et al., 2004), They are also
5
of interest to biologists and clinicians because they may help
to protect the human body against damage by reactive oxygen
species (ROS). ROS is a collective term used to include oxy-
gen radicals (O− • •
2 , OH , RO2 , RO , etc.) and several non-radical
oxidizing agents such as HOCl (hypochlorous acid), H2 O2 , O3
(ozone) (Halliwell, 1991). In biological systems, lipid peroxi-
dation can occur mainly in biomembranes, where the content of
unsaturated fatty acids is relatively high.
SYNTHETIC ANTIOXIDANTS
Synthetic antioxidants are mainly phenolic and include buty-
lated hydroxyanisole (BHA), butylated hydroxy toluene (BHT),
tertiary butyl hydroquinone (TBHQ) and propyl, octyl, and do-
decyl gallates.
The use of phenolic antioxidants in foods had its beginning
in the late 1940s when BHA was found to be effective antioxi-
dants in fatty foods and toxicological studies proved it safe for
food use. Later BHT was approved in the United States and put
to widespread use along with previously available food antioxi-
dants. In 1972, TBHQ was commercialized as a food-grade phe-
nolic antioxidant for polyunsaturated oils, such as safflower seed
oil, extensively used in baked products (Sherwin, 1989). Poly-
meric antioxidants such as Anoxomer, Ionox-330, and Ionox-
100, a derivative of BHT, have also been introduced, but they
are not being used commercially (Joyner and McIntyre, 1938).
Antioxidants for use in baked foods must satisfy the follow-
ing requirements such as they must be established as substance
having a very low order of toxicity, they should not affect the
odor, flavor, or appearance of the end product, they should be
effective at low concentrations under normal conditions of pro-
cessing and storage of the end product, they must be stable at
high temperature for certain applications such as frying and bak-
ing, and the antioxidant should be soluble and dispersible in the
end product, in emulsions and other preparations, that contain
water.
The action of phenolic compounds (Fig. 2) in inhibiting free-
radical autooxidation may be depicted simply by showing that
the phenol (serving as a proton donor) suppresses initial free rad-
ical (R• ) formation, thus delaying the onset of the autoxidative
process in fat or oil (RH). The antioxidant free radical formed
in this reaction, unlike a fatty acid free radical, does not have
the capability of initiating or propagating oxidation of the fat
or oil. This serves to demonstrate that autooxidation is not pre-
vented, but its onset is just delayed and the extent of the delay
is dependent on the activity of the particular antioxidant and its
concentration as well as other factors such as heat, light, metals,
and other pro oxidants in the system (Sherwin, 1989).
Butylated Hydroxyanisole (BHA)
Butylated hydroxyanisole (BHA; tertiary butyl-4-
hydroxyansole) is one of the extensively used food
 6 B. NANDITHA AND P. PRABHASANKAR

OH OH stabilization of essential oils like d-limonene, orange oil, lemon

oil, and terpenes is widely carried out using BHA.

+ RH
R'
OH
Applications in Bakery Products
The most important property of BHA is its ability to remain
OH
O active in baked or fried foods. BHA finds widespread use in low
fat foods such as cereal products, especially breakfast cereals and
O OH cake mixes. The stability of pastry and cracker ranged from 27–
O 33 days in the presence of 0.02% BHA respectively (Madhavi
R' et al., 1996). At 0.01% level, both products were stable up to
21–22 days.
The volatility of BHA and BHT is an advantageous property
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in low fat foods. There are many ways of adding of BHA to the
O O
foods, one of them being adding to the potato or cereal slurry
Figure 2 Mechanism of action of phenolic antioxidants.
before cooking or which drying which results in dispersion by
volatilization or steam distillation, resulting in the protection of
the product during processing and subsequent storage. Another
antioxidants. Commercial BHA is a mixture of two iso-
approach has been to spray an emulsion of the antioxidant onto
mers, 2-tert-butyl-4-hydroxyanisole (2-BHA, 10%), and 3-tert-
the finished product immediately prior to packaging. BHA used
butyl-4-hydroxanisole (3-BHA, 90%) (Fig. 3a and b). BHA
in bakery products at levels of 0.01–0.04% (Joyner and McIn-
is highly soluble in fats and oils and insoluble in water. BHA
tyre, 1938).
may also develop a pink color in the presence of alkaline metals
in the baked product (Joyner and McIntyre, 1938). BHA has
low melting point and is steam volatile at frying temperatures. Anti-Microbial Activity of BHA
However, the residual BHA exhibits considerable carry-through
BHA is found to have a profound antimicrobial effect against
properties in baked and fried foods (Madhavi et al., 1996.).
Aspergillus parasiticus, Staphylococcus aureus, Escherichia
BHA is a “hindered” phenol, since the tert-butyl group ortho
coli and Salmonella typhimurium (Chang and Branen, 1975).
or meta to the hydroxyl group suppresses antioxidant activity.
Antifungal properties of BHA against species of Aspergillus,
BHA is a white, waxy solid that is usually tableted to minimize
Penicillium, and Geotrichum in synthetic media and food prod-
caking during storage.
ucts were found (Ahmed and Branen , 1981). Rapid bactericidal
Commercially BHA contains a high proportion of the 3-
action of BHA against a number of organisms was observed,
isomer (90%) which is nearly as effective as pure 3-BHA mainly
particularly a high level of activity against Pseudomonas aerug-
because of a slight synergism between the two isomers. The ad-
inosa, an organism resistant to a wide variety of antibacterials.
dition of BHA causes no change in color, odor, or flavor. It works
The antibacterial effects of BHA against Bacillus species are
well when mixed with other antioxidants such as BHT, TBHQ,
studied by Shelf and Liang (1982). It was found that the growth
or Propyl gallate (PG), and the resulting synergism and pro-
inhibition level of BHA was 75 ppm on enterotoxigenic strains
vides greater antioxidant potency than that might be expected
of B. cereus, toxigenic strains of B. cereus, B. subtilis, and B.
from the contribution of each individual antioxidant. BHA is a
megaterium in nutrient broth. By using disc diffusion and plate
popular antioxidant choice for breakfast cereals, vegetable oils,
count methods it is found that BHA at 2,500 ppm was effective
active dry yeast, baked goods etc. (Coulter, 1988). BHA (0.01%)
in completely inhibiting the growth of Aspergillus flavus and
mixed with dodecyl gallate (0.005%) is more effective than BHA
Aspergillus fumigatus. BHA at 250–ppm level totally inhibited
alone in stabilizing margarine (Tollenaar and Vos, 1958). The
growth and toxin production of A. parasiticus in glucose salt
medium. The antimicrobial activity of BHA depends on the pH
OH CH3 OH of the medium.
CH3

CH3 CH3 Retention of BHA after Processing

CH3 The main purpose of adding an antioxidant is served only
O
if the added antioxidant is retained in the product after it is
O CH3
H3C H3C processed. Many research studies were conducted by various re-
(b) searchers in this context. Behavior of antioxidants during baking
(a)
and storage of Pie Crust was studied by Mahon and chapman
Figure 3 Isomers of Butylated hydroxy anisole (BHA) (a) 2-BHA (b) 3-BHA. (1953) and it was observed that recoveries of BHA from dough
 ANTIOXIDANTS IN BAKERY PRODUCTS
have been as high as 98% though in most instances recoveries
from 80 to 92% were obtained. In case of piecrust the time of
baking normally employed resulted in a loss of 15 to 20% of the
BHA present. The loss of BHA proceeds at an ever-increasing
rate during the baking process. The loss of BHA was due at
least in part to the steam-distillation of BHA from the dough.
There was approximately 15% loss of BHA during the baking
process. Thereafter, the loss of total BHA, 2-BHA, and 3-BHA
approximated a straight-line function of the time of storage at
61◦ C.
In another study the fate of antioxidants and antioxidant-
derived products in deep fat frying and cookie baking were re-
vealed by radioactively labeling the antioxidants as ring labeled
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[14 C] BHA (2-(1,1-dimethlethyl)-4-methoxyphenol) and ring-
labeled [14 C] TBHQ (2-(1,1-dimethlethyl)-1,4-benzenediol)
(Warner et al., 1986).
Permitted Levels of BHA in Bakery Products
BHA is Generally Recognized as Safe (GRAS) for use in
food when the total content of antioxidants is not over 0.02% of
the fat or oil content (including the essential (volatile) oil con-
tent) of food, provided the substance is used in accordance with
good manufacturing practice (Coulter, 1988). It may be used as
an antioxidant in flavoring substances in amounts not exceed-
ing 0.5% of the essential (volatile) oil content of the flavoring
substance. BHA is approved for food use by most of the coun-
tries except Greece, Jamaica, and Turkey. The Allowable Daily
Intake (ADI) for BHA is 0–0.5 mg/kg body weight.
Toxicology of BHA
Toxicological studies are crucial in determining the safety of
an antioxidant for food use and also in determining the Allow-
able Daily Intake (ADI) levels. The absorption and metabolism
of BHA has been studied in rats, rabbits, dogs, monkeys, and
humans. BHA was rapidly absorbed, from the gastrointestinal
tract of rats, rabbits, dogs, and humans, rapidly metabolized,
and excreted completely. The major metabolites of BHA were
the glucuronides, ether sulfates and free phenols. The acute oral
LD50 was 2200–5000 mg/kg body weight in rats and 2000 mg/kg
body weight in mice. In high doses (500 mg/kg body weight per
day), BHA induced an increase in the relative liver weight in rats
and mice. Administration of BHA at 2% level resulted in high
incidence of papilloma in almost 100% of the treated animals
and squamous cell carcinoma of the fore stomach in about 30%
of the treated animals. Among the BHA isomers, severe adverse
effects were observed with crude BHA and the 3-isomer, but the
2-isomer had no effect (Joyner and McIntyre, 1938). In rodents,
hypertrophy of the liver has been the most common pathologi-
cal change, which the occurred with the administration of BHA
(Branen, 1975).
7
CH3 OH CH3
H3C CH3
CH3 CH3
CH3
Figure 4 Butylated Hydroxy Toluene.
Butylated Hydroxytoluene (BHT)
Butylated hydroxytoluene (BHT; 2,6-di-tert-butyl-p-cresol
or 2,6, di-tert-butyl-4-methylphenol) (Fig. 4) is also extensively
used in food products and is similar to BHA in many of its prop-
erties. BHT is a white crystalline solid with a faint phenolic odor.
BHT is highly soluble in fats and oils and insoluble in water. It is
insoluble in propylene glycol and moderately soluble in glyceryl
monooleate, a solvent used for commercial antioxidant formula-
tions. However, BHT is not as effective as BHA mainly because
of the presence of two tert-butyl groups, which offer greater
steric hindrance than BHA. It is more steam volatile than BHA
and has less carry-through properties than BHA. It offers fairly
good carry-through after baking and frying to protect the final
cooked product (Joyner and McIntyre, 1938). BHT also forms
a yellow color in the presence of iron ions in food products or
packaging materials.
The addition of BHT causes no changes in color, odor, or
flavor of a food product. It may be used in combination with
BHA, propyl gallate or TBHQ. Stillson patented BHT in 1947
for usage in animal fat and dry breakfast cereals (Adegoke et al.,
1998).
Applications in Bakery Products
BHT resembles BHA in many of its antioxidant properties
and is used in similar food applications. BHT functions syner-
gistically with BHA, TBHQ, and chelators like citric acid and
have no synergistic activity with propyl gallate. BHT does not
have an optimum concentration, and the stability of fats to which
it is added continues to increase with concentration. At concen-
trations higher than 0.02%, BHT imparts a phenolic odor to the
fats (Joyner and McIntyre, 1938).
BHT is most frequently used as an antioxidant for animal
fats, dry breakfast cereals, food packaging materials, potato
granules and flakes, vegetable oils, snack foods, baked goods
etc. (Coulter, 1988). BHT is more effective than BHA for sta-
bilizing animal fats and is used at a concentrations of 0.005–
0.02%. Table 2 gives the level of BHT used in various food
systems. The addition of BHT to pastry and crackers has signif-
icantly increased the shelf life of the products (Madhavi et al.,
1996).
 8 B. NANDITHA AND P. PRABHASANKAR
Table 2 Levels of BHT used in practice in food products∗
Product Level (%)
Vegetable oils 0.002–0.02
Bakery products 0.01–0.04a
Cereals 0.005–0.02
a Calculated on the fat.
∗ Reference : (Madhavi et al., 1996).
Permitted Levels of BHT in Bakery Products
BHT is Generally Recognized as Safe (GRAS) for use in
food when the total content of antioxidants is not over 0.02%
of the fat or oil content (including the essential (volatile) oil
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content) of food, provided the substance is used in accordance
with good manufacturing practice (21 CFR 182.3173) (Coulter,
1988). ADI for BHT is 0–0.125 mg/kg body weight. In cake
mixes BHT should not exceed 200 mg/kg (gallates and BHA,
individually or in combination).
Toxicology of BHT
The metabolism of BHT is more complicated and slower
than that of BHA. The relative slow excretion of BHT has been
attributed to the enterohepatic circulation. Rats are given with 0.5
and 1% BHT for 5 weeks and the concentration of BHT increased
rapidly in liver and body fat. The acute oral LD50 in mg/kg body
weight in rats was 1700–1970: in rabbits, 2100–3200. In rats,
BHT at the level of 0.3 or 0.5% caused an increase in the level
of serum cholesterol and phospholipids within 5 weeks. In high
doses, BHT had a toxic effect on liver, lung, and kidney and also
on the blood coagulation mechanism leading to hemorrhages.
ADI values for BHT have changed over the years because of its
toxicological effects in different species. The latest temporary
value allocated by Joint Expert Committee on Food Additives
(JECFA) is 0–0.125 mg kg body weight (FAO/WHO, 1987).
Other changes include diminished initial growth rate and loss of
hair of rats fed with BHT. BHT has caused skin irritation in a
test using guinea pigs but is not recognized as causing irritation
after oral ingestion at current allowable intake levels. BHT is
not recognized as a reproductive or developmental toxicant.
Degradation of BHT
Antioxidants undergo degradation during autooxidation of
fats and oils. The study of degradation helps in understand-
ing the mechanism of action of primary antioxidants, proper-
ties of the degradation products, and the role of synergists in
regenerating the primary antioxidants. Studies on the degra-
dation of BHT in soybean oil irradiated by UV light showed
that the concentration of BHT decreased and completely lost
at the end of the induction period of the oil. The degrada-
tion products have been identified as BHT aldehyde (3,5-di-
tert-butyl-4-hydroxybenzaldehyde), BHT alcohol (3,5-di-tert-
butyl-4-hydroxybenzyl alcohol), butylbenzoquinone (BQ), and
diphenylethylene (BE).
Synergism between BHA and BHT
Synergists function by a different mechanism for extension
of the shelf life of a primary antioxidant and it is found that the
antioxidant effect of the mixture is greater than when they are
used alone. Synergists may act as hydrogen donors to the primary
antioxidant radicals, thus regenerating the primary antioxidant
or inactivate metal ions, thus neutralizing their prooxidant ef-
fects. It was observed that 3-BHA donated a hydrogen atom to
peroxy radical to form a phenoxy radical. A transfer of hydro-
gen from BHT regenerated 3-BHA from the phenoxy radical. In
the process BHT was oxidized to quinone methide (Fig. 5). A
combination of 3-BHA and BHT showed a higher antioxidant
activity than either of them used singly in soybean oil, lard, and
methyl oleate (Madhavi et al., 1996).
Tertiary Butyl Hydroquinone (TBHQ)
TBHQ (Fig. 6) was approved for food use in 1972. TBHQ is
very effective in stabilizing fats and oils, especially polyunsat-
urated crude vegetable oils. The two parahydroxyl groups are
believed to be responsible for its antioxidant activity. TBHQ is
stable in high temperatures and is less steam volatile than BHA
and BHT. TBHQ is a white or light tan crystalline powder that
is slightly soluble in water (<1%) and moderate by soluble in
fats and oils (5–10%). It does not form a complex with iron or
copper.
Applications in Bakery Products
The activity of TBHQ is equivalent to or greater than that of
BHA, BHT, or PG. TBHQ does not contribute to the color and
odor of fats and oils. TBHQ functions synergistically with other
antioxidants and chelators such as PG, BHT, BHA, tocopherols,
ascorbyl palmitate, and citric acid. The most interesting property
of TBHQ is its effectiveness in various oils and fats where other
phenolic antioxidants are relatively ineffective. There are indi-
cations that TBHQ will interact with free amines to give a red
coloration that may preclude its use in some proteinaceous foods
where such coloration is not tolerable (Sherwin, 1989). Studies
indicated that TBHQ does not survive the processing conditions
involved in the preparation of pastry and crackers. Hence, TBHQ
does not carry through into the baked product. But TBHQ has a
significant carry-through effect in frying operations.
Toxicology of TBHQ
The Joint Expert Committee on Food Additives (JECFA) allo-
cated a temporary Allowably Daily Intake (ADI) of 0–0.2 mg/kg
body weight. Absorption and metabolism studies have been con-
ducted in rats, dogs, and humans and it is found that more than
90% of an orally administered dose of TBHQ was rapidly ab-
sorbed and nearly 80% was excreted in the urine in the first
24hr. The acute oral LD50 in mg/kg body weight in rats was
700–1000. At a dose of 0.5% level a reduction in food intake
 ANTIOXIDANTS IN BAKERY PRODUCTS 9

OH CH3 O. CH3

CH3 CH3
ROO.
CH3 CH3
+ ROOH

O O
H3C H3C
Phenoxy radical
BHA
H2

CH3 O CH3 CH3 CH3

OH
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H3C CH3 H3C CH3

CH3 CH3 CH3 CH3

CH2 CH3

Quinone methide BHT

Figure 5 Synergism between BHA and BHT.

with a consequent decrease in the body weight of the pups was
observed. TBHQ had no effect on reproductive system and was
not teratogenic. Results on mutagenicity indicated that at higher
levels, TBHQ might be mutagenic.
Antimicrobial Activity of TBHQ
TBHQ at the rate of 0.01% was found to inhibit aerobic spore
formers for soy flakes in Standard Plate Count agar. TBHQ (5–10
μg/ml) inhibited lactic acid production by P. pentosaceus. The
growth of S. cerevisiae in ice cream mix was retarded by TBHQ
(500 ppm). TBHQ decreased the incorporation of 14 C acetate
into lipids and proteins of T. pyriformis and also inhibited the
incorporation of 14 C amino acid into the proteins of the same
organism (Raccah, 1984). TBHQ was effective in preventing the
growth of S. aureus in a nutrient broth. Other studies showed
that TBHQ was extremely effective against gram +ve bacteria.
TBHQ at 600 ppm level inhibited Aspergillus Niger and at 400
ppm level inhibited Penicillium citrinum.
OH CH3
CH3
CH3
Gallates
Gallic acid (3,4,5-trihydroxybenzoic acid) is widely dis-
tributed in nature, mostly as a main component of tannins and
occurring freely in tea and pomegranates. It was shown to be
an effective antioxidant as well as synergist for other antioxi-
dants, but it is soluble in water and virtually insoluble in fats.
The Gallates used as food antioxidants include propyl, octyl,
and dodecyl esters of gallic acid (Fig. 7). The gallate esters
are all white or creamy white solids. Propyl gallate is one of
the most widely used food antioxidants and is a component of
many commercial antioxidant mixtures. It is less soluble in fats
and oils than BHA and BHT and has significant solubility in
water.
The dodecyl and octyl esters are more soluble than propyl
gallate in fats and oils. In general, the gallates do not have sig-
nificant carry-through properties. Of the three gallates, propyl
gallate is most sensitive to heat and undergoes degradation at
frying temperatures. The temperature stability improves with in-
crease in molecular weight that is octyl and dodecyl gallates are
more stable to heat and possess better carry-though properties.
Propyl gallate forms purple or violet complexes with iron ions,
resulting in discoloration of the food product. Hence, propyl
gallate is always used in combination with metal chelators such
as citric acid. Propyl gallate was approved for food use by the
Food and Drug Administration in 1947 (Joyner and McIntyre,
1938).

Applications in Bakery Products

OH
Gallates are widely used in fats and oils, meat products, con-
Figure 6 Tertiary Butyl Hydroxy quinone. fectioneries and nuts, milk products, fish products, margarine,
 10 B. NANDITHA AND P. PRABHASANKAR

O O O O CH3
CH3

HO OH
HO OH
OH
OH
(a) (b)

O O CH3
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HO OH
OH

(C)

Figure 7 (a) Propyl gallate (b) Octyl gallate and (c) Dodecyl gallate.

and in bakery products. In baked goods a combination of BHA Toxicology of Gallates

and Dodecyl Gallate (DG) is effective. Table 3 gives an idea of
the level of gallates used in various food products either individ-
ually or in combination with other antioxidants such as BHA,
BHT etc.
In bakery products the result varied considerably although in
some cases the incorporation of 0.05% dodecyl gallate, (calcu-
lated on the fat) was found to exercise a favorable influence on
the flavor.
Retention of Gallates after Processing
Behavior and retention of antioxidants during baking and
storage of Piecrust was studied and it was found that the loss
of lauryl gallate in the presence of citric acid is a straight-line
function of storage time at 61◦ C. In the case of piecrust made
from dough containing “Versene” and then sprayed with lauryl
gallate plus citric acid, the loss of lauryl gallate approximated a
straight-line function of time (Mahon and Chapman, 1953).
Table 3 Levels of Gallates used in various food products∗
JECFA allocated an Allowable Daily Intake (ADI) of 0–
0.25 mg/kg body weight for propyl gallate and no ADIs were
allocated to octyl gallate (OG) and dodecyl gallate (DG) because
of insufficient toxicological information (Branen, 1975). In rats,
nearly 70% of an oral dose of PG was absorbed in the gastroin-
testinal tract whereas OG and DG were absorbed to a lesser
extent. All three esters were hydrolyzed to gallic acid and free
alcohol. The acute oral LD50 of PG in rats was 3000–3800 mg/kg
body weight, for OG was 4700 mg/kg body weight, and for DG
it was 6500 mg/kg body weight. In a long-term study, rats fed 5%
PG showed reduced food intake, growth retardation, and patchy
hyperplasia of the stomach. All the three gallates were found to
have no adverse effects on reproduction in a three-generation
study in rats. The gallates have been reported to cause allergic
contact dermatitis. Propyl gallate has been reported to be neg-
ative in a number of mutagenicity tests. All three esters were
hydrolyzed to gallic acid and free alcohol (Fig. 8). Gallic acid
was methylated to 4-O-methyl gallic acid, which was found in
urine (Madhavi et al., 1996).

Product Level (%)a

Vegetable oils 0.001–0.01, all gallates DETERMINATION OF ANTIOXIDANTS IN FOOD

Whole milk powder 0.001–0.02, all gallates
Margarine 0.001–0.01, all gallates
SYSTEMS
Bakery products 0.01–0.04, mainly DGb
A large number of antioxidants are used in food systems es-
DG= dodecyl gallate.
a Combination with BHA, BHT and citric acid are frequently used. pecially bakery products for the purpose of increasing the shelf
b Calculated on the fat. life by retarding the oxidation of fats. Determination of antiox-
∗ Reference:(Madhavi et al., 1996).
idants is of prime importance in view of legal specifications,
 ANTIOXIDANTS IN BAKERY PRODUCTS 11

COOH COO Gluc

COOR
COOH

HO OH HO OH

HO OH O O
HO OH H3C H3C
OH OH 4-O- Methyl gallic acid
Gallic acid

+
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ROH HO OH
HO OH
OH O Gluc

Pyrogallol

Where R = C3H7 : Propyl gallate

C8H17: Octyl gallate
C12H25: Dodecyl gallate

Figure 8 Major metabolites of the gallates.

stability, effectiveness, consumer preference, and cost. Various
methods are available for their determination depending on the
type of antioxidant and the type of food product.
Extraction
Primary antioxidants such as BHA, BHT, and the gallates are
extracted from bakery products with either aqueous alcohol or
acetonitrile. Aqueous alcoholic extraction of antioxidants from
bakery products has the disadvantage that losses by volatiliza-
tion of BHA and BHT are likely to occur if a concentration step
is incorporated. Hence extraction with acetonitrile or dry alcohol
or a combination of the two is employed. Propyl gallate can be
extracted with water from a solution of a fat-antioxidant mixture
in petroleum ether, hexane or heptane. A clean-up step is gen-
erally incorporated to remove the interfering substances before
analysis. Column chromatography using polyamide, silica gel,
or florisil is generally employed, depending on the antioxidant
in the extract.
The methods for determination of antioxidants in food system
are as follows (AOAC, 1984; Rafecas et al., 1998).
•
Spectroscopic Methods
•
Chromatographic Techniques.
in bakery products. After the sample is oxidized under standard
conditions, the extent of oxidation is measured by chemical, in-
strumental, or sensory methods. Most of these studies are aimed
at measuring the extension of the induction period by the ad-
dition of the antioxidant. The extension in induction the period
is also expressed as an antioxidant index or protection factor.
The induction period is the time required for the sample to start
oxidizing rapidly and coincides with the onset of off-flavor de-
velopment in the lipids.
• Accelerated Stability Methods
• These methods generally involve storage studies under normal
or accelerated conditions. Some of the conventional stability
tests are listed in Table 4 along with their characteristics.
• Improved Techniques of Oxygen Bomb Method (Stuckey
et al., 1958)
• Automated Active Oxygen Method (AOM) Test (Whetsel
et al., 1957)
Measurement of Oxidative Stability
The formation of hydroperoxides or their decomposition
products is measured by a chemical, instrumental, or sensory
method. The analytical methods to measure oxidative stability
of fats and oils are listed in Table 5 (Joyner and McIntyre, 1938).

Determination of Antioxidant Activity

Infrared Analysis of Commercial BHA
The effectiveness of synthetic and natural antioxidants is An infrared method has been developed which allows the
measured by monitoring the oxidative stability of the food lipids isomers to be determined with a standard deviation of approxi-
 12 B. NANDITHA AND P. PRABHASANKAR

Table 4 Standard Accelerated Stability Tests∗

Test Conditions Characteristics

Ambient storage Room temperature, atmospheric pressure Too slow

Light Room temperature, atmospheric pressure Different mechanism
Metal catalysts Room temperature, atmospheric pressure More decomposition
Weight-gain method 30–80◦ C, atmospheric pressure Endpoint questionable
Schall oven 60–70◦ C, atmospheric pressure Fewest problems
Oxygen uptake 80–100◦ C, atmospheric pressure Different mechanism
Oxygen bomb (ASTM)a 99◦ C, 65–110 psi O2 Different mechanism
Active oxygen (AOM) 98◦ C, air bubbling Different mechanism
Rancid at 100–140◦ C Endpoint questionnable
a ASTM= American Society for Testing and Materials.
∗ Reference:
(Madhavi et al., 1996).
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mately ±1%. The method is rapid and requires very little recal- oped enormously in the past decade mainly because of the in-
ibration and has been used for several years as a convenient and creasing limitations on the use of synthetic antioxidants, and en-
effective quality control measure on commercial butylated hy- hanced public awareness of health issues. Recent studies shows
droxyanisole. This test is highly recommended if infrared equip- that these synthetic antioxidants possess toxic, pathogenic and
ment is available. In this method, the ratio of the absorbances at carcinogenic effects, effect on enzymes, lipids, effect on repro-
10.74 and 10.92 microns of a carbon disulfide solution of buty- duction. Hence, there is an increasing need for health-promoting
lated hydroxyanisole provided a reliable value for the isomeric natural antioxidants such as β-carotene, α-tocopherol, ascorbic
composition of the commercial product. Strong bands charac- acid, phenols, terpenoids etc. Advantages and disadvantages of
terize the 3-BHA at 9.45, 13.02, and 13.22 microns while the both synthetic and natural antioxidants are given in Table 6.
2-BHA isomer shows distinctive band sat 9.65, 10.74, and 13.35 Even though there are some disadvantages in the application of
microns. Each of the three samples of BHA containing approxi- natural antioxidants they are invariable alternatives to synthetic
mately 99, 93, and 80% of the 3-BHA isomer was analyzed eight antioxidants. Further research is needed in this direction.
times by two laboratory technicians over a 2 month period. The A wide range of natural antioxidants has been shown to occur
standard deviations for the isomer determinations were 0.4, 0.5, in plants and animals. Although these compounds can be syn-
and 1.2% respectively (Whetsel et al., 1957). thesized, they are treated as if they belong to natural sources. In
recent years, there has been a greater awareness of use of nat-
ural antioxidants and the promotion over the typical synthetic
NATURAL ANTIOXIDANTS antioxidants such as BHA and BHT. Natural antioxidants at-
tracted the attention of many food manufacturers as a result of
necessity created by a situation where it was extremely difficult
Need for Natural Antioxidants
to use synthetic antioxidants especially for developing health
foods. Moreover, new toxicological data on some of the syn-
Since the importance of antioxidants was found they are ex-
thetic antioxidants cautioned against their use. In addition to
tensively used in food products. In 1967, 15.5 million lb an-
their long-term safety and capacity to improve food quality and
tioxidants valued at $20.5 million were used by food manu-
stability, these natural antioxidants may also act indirectly as
facturers of the U.S. (Branen, 1975). Although many antioxi-
nutraceuticals by terminating free radical chain reactions in bio-
dants are available, most antioxidant formulations contain BHA
logical systems and therefore may provide additional nutritional
and BHT, which are synthetic in nature. In recent years, con-
benefits to consumers.
sumers, and food manufacturers have been opting for products
However, it is difficult to formulate a safe procedure for the
with “all natural” labels. The area of natural antioxidants devel-
use of either synthetic or natural antioxidants in food systems.
There exist a wide range of natural antioxidants some of which
Table 5 Analytical methods to measure oxidative stability of fats and oils∗
are currently used effectively for the protection of foods against
Chemical methods Chromatographic methods oxidative degradation. The mechanism of action of synthetic
Peroxide value HPLC and natural chain-breaking antioxidants is similar. Both of them
Anisidine value Gas chromatography act as radical scavengers and donate hydrogen atoms to reduce
Thiobarbituric acid test GC-MS
by a one-electron transfer reaction the primary radicals formed
Kreis test Measurement of oxygen absorption
Spectrometric methods Dissolved oxygen meter during the autooxidation reactions.
UV absorption Warburg’s manometer Numerous chemicals that are found in animal or plant tissues
Carotene bleaching test Sensory methods are used in several food applications (Tables 7 and 8). Among
ESR spectrometry Flavor and odour evaluation these chemicals, vitamin E, vitamin C, β-carotene, and uric acid
IR spectrometry
are very interesting synthetic products of natural origin capable
∗ Reference: (Madhavi et al., 1996; Niki, 1996). of participating in the in vivo radical defense mechanism. The
 ANTIOXIDANTS IN BAKERY PRODUCTS 13

Table 6 Comparison of natural antioxidants with synthetic antioxidants∗

Synthetic Antioxidants
Advantages Disadvantages
Machanism of action and their use is well established. More efficient. Showed positive results for some toxicological studies.
Comparatively Cheaper May impart color, aftertaste, and flavor.
Stable at processing conditions of temperature and time. Not readily accepted by consumer, as they are synthetic compounds.
Easy purification steps. Their use is regulated by PFA act.
Natural Antioxidants
Readily accepted by the consumer, as considered to be safe. Usually more expensive if purified
Considered as safe for consumption and are under “generally recognized Properties of different preparations vary if not purified
as safe” (GRAS).
No positive results for toxicological studies as on now. Mechanism of action is not well established.
Along with antioxidant property also serves other purposes such as coloring Not stable under high temperature and time combinations of processing.
agent in case of ß-carotene.
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∗ Reference: (Madhavi et al., 1996; Niki, 1996)

vitamin E and vitamin C mixes became the basis for several sues, especially in nuts, vegetable oils, fruits, and vegetables.
approaches to the stabilization of oils and other foods because Some of the rich sources are wheat germ, corn, sunflower seed,
of the radical exchange reactions between lipid radicals, vitamin rapeseed, soybean oil, alfalfa, and lettuce. All the compounds
E, and vitamin C. exhibit antioxidant and vitamin E activities. However, they have
Besides biologically active compounds, pure chemical sub- limited carry-through properties and are less effective than BHA,
stances of plant origin treated as part of the daily food intake BHT, and PG. The tocols and tocotrienols include α, β, γ and δ
can be added as stabilizing agents against oxidative degrada- homologs. The basic structural unit in all the homologs is a 6-
tion. Amino acids, which are responsible for producing Mail- chromanol ring with a phytol side chain (Fig. 9). The homologs
lard reaction products, belong to this group. Vanillin, the most differ in the number of methyl groups bound to the aromatic ring.
widely used food-grade flavoring agent, has been recently rec- The tocotrienols differ from tocols in having an unsaturated side
ognized for its antioxidant potential. Plant extracts contain a chain, with double bonds at the 3 , 7 , and 11 positions.
variety of natural products, but certain active principles are re- The tocols are generally referred to as tocopherols. The an-
sponsible for their antioxidant activity. Oats are an easily avail- tioxidant activity of the α, β, γ , and δ homologs varies with
able source of antioxidants and appear to be promising in low the degree of hindrance and temperature. At physiological con-
concentrations for effective retardation of the rate of oxidation. ditions around 37◦ C, the antioxidant activity is in the order
Tea antioxidants could prove to be useful as natural antioxidants α > β > γ > δ, similar to the biological activity. At higher
if large-scale applications become successful. Herbs and spices temperatures of 50–100◦ C, a reversal in the order of antioxi-
occupy a special position in foods as traditional food ingredients dant activity is observed as δ > γ > β > α (Madhavi et al.,
and hence are appropriately used directly for their antioxidant 1996).
characteristics. α-Tocopherol is the most abundant of all the tocopherols and
its biological activity is twice that of β, and γ homologs and 100
times that of the δ homolog (Hix et al., 1997). Tocopherols are
Tocopherols pale yellow, clear, viscous, oily substances, insoluble in water
and soluble in fats and oils.
The tocopherols comprise of a group of chemically related α-Tocopherol and its acetate have also been chemically syn-
tocols and tocotrienols that are widely distributed in plant tis- thesized. Synthetic products are designated as dl-α-tocopherol
and dl-α-tocopheryl acetate, which are actually mixtures of the
Table 7 Some naturally occurring antioxidants∗
four racemates. A synthetic water soluble form of α-tocopherol,
d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), has
Amino acids β-Carotene been prepared by the etherification of d-α-tocopheryl acid suc-
Carnosine Carnosol
cinate with polyethylene glycol having an average molecular
Citric acid Curcumin
Eugenol Flavonoids weight of 1000. It is a pale yellow, waxy substance that provides
Lecithin Lignans 260 mg of d-α-tocopheryl per gram and forms a clear solution
Nordihydroguaiaretic acid Phenolic acid in water at concentration up to 20% (Johnson, 1971; Krasaveges
Phytic acid Protein hydrolysates and Terhaar, 1971).
Proteins Rosmarinic acid
Saponins Sterols
Tartaric acid Uric acid Application in Bakery Products
Turmerin Vitamin C
Vitamin E Vanillin Tocopherols are considered the major natural antioxidants
∗ Reference: in vegetable and animal fats. At the concentrations necessary
(Madhavi et al., 1996).
 14 B. NANDITHA AND P. PRABHASANKAR

R'' CH3

HO O CH3 CH3 CH3

CH3
CH3
R'
R' R"

alpha-tocopherol -CH3 -CH3

beta-tocopherol -CH3 -H

gamma-tocopherol -H -CH3
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delta-tocopherl -H -H
R'' CH3

HO O CH3 CH3 CH3

CH3
CH3
R'
To cotrienols

Figure 9 Tocopherol and Tocotrienol.

to give adequate protection to low-stability fats, they usually
contribute an undesirable foreign flavor of their own (Leenhardt
et al., 2006). The tocopherols function synergistically with other
antioxidants such as BHA, TBHQ, ascorbyl palmitate, lecithin,
amino acids, ascorbic acid and various spice extracts. The use
of tocopherols is limited to fats, oils, and fat-containing foods.
The tocopherols have carry-through properties in baked and
fried products. Crackers and pastry prepared with lard treated
with 0.01–0.1% tocopherol, either alone or in combination with
BHA (0.01%), were appreciably more resistant to rancidity than
control samples (Chipault, 1962; Dugan and Kraybiu, 1962;
Dugan and Kraybill, 1956).
Hix et al. (1997) conducted a study on physical and chemical
attributes and consumer acceptance of sugar-snap cookies con-
taining naturally occurring antioxidants. The objective of this
study was to test the shelf life and other properties of sugar-snap

Table 8 Potential sources of natural antioxidants∗

Plant extracts
Cocoa shells Leaf lipids Oats
Tea Olive Garlic
Red onion skin Wheat gliadin Apple cuticle
Korum rind Licorice Nutmeg
Mustard leaf seed Chia seed Peanut seed coat
Rice hull Birch bark Carob pod
Myristica fragrans Silbum marianum seed oil Carob pod
Spices and herbs
Rosemary Pepper Clove
Oregano Spices, general
Chemicals in animal and plant tissues
Vitamin E Vitamin C β-Carotene
Uric acid Phenolic acids L-Ascorbic acid ester
Phytic acid Amino acids Sesame seed oil
Flavones Saponin Soya Saponins
Vanillin Nor dihydro guaritic acid Osbeckia chinensis tannins
Fermentation products
Penicillium commune Tempeh oil Microorganisms
Penicillium herquei
Food components or products of heat-processed foods
Peptide and sugar Phospholipids Amino acids
Rosmarinic acid and phospholipids Ascorbyl palmitate, lecithin, tocopherols Maillard reaction products
Soy protein hydrolysates
∗ Reference : (Madhavi et al., 1996; Pokorny and Korezak, 1996).
 ANTIOXIDANTS IN BAKERY PRODUCTS 15

cookies containing ascorbate, α-tocopherol, sodium phytate, and Table 9 Total Tocol isomers contents before (whole wheat flours)
ferulic acid as viable replacements for BHA. The cookies were and after bread making (whole wheat breads, expressed in μg/g of
dry weight)∗
compared with regard to shelf life, moisture content, and width,
stacking height, surface score, color, and texture. Following Tocopherol (μg/g dw) Tocotrienol (μg/g dw)
storage at 60◦ C, the Schaal oven test and gas-liquid chromato- α β α β
graphic analysis of headspace gases indicated that ferulic acid
and sodium phytate were suitable antioxidants for sugar-snap T. monococcum
Whole flour 11.03 7.78 4.87 40.94
cookies. In no case the test cookies were different than the BHA Whole bread 9.17 6.16 3.02 14.83
control cookies. Hence, from the experiment it can be under- T. durum
stood that natural antioxidants can be effectively substituted for Whole flour 11.52 6.93 6.81 31.41
BHA in sugar-snap cookies (Hix et al., 1997). Whole bread 9.46 5.43 3.75 7.57
Park et al. (1997) studied the effect of addition of fiber ingre- T. aestivum
Whole flour 14.93 6.24 9.7 44.97
dients with different antioxidants on bread characteristics. The Whole bread 12.24 5.49 5.11 13.7
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results indicated that the addition of those fiber ingredients in-

∗ Reference:
creased dough absorption ≈25% and mixing time by ≈50%.The (Niki, 1996).
fiber and antioxidant bread showed a 10% reduction in loaf vol-
ume and a somewhat inferior crumb grain with an off-color
caused by small, black specks on a dark gray background (Park
et al., 1997). No further loss of ToAc occurred during baking and 7 days of
storage (Park et al., 1997).
Wennermark and Jagerstad (1992) conducted a study on the
Retention of α-Tocopherol after Processing effect of breadmaking on vitamin E. The vitamin E loss during
baking of two types of industrially baked breads was studied.
In an experiment conducted by Park et al. (1997) 10% weight Neither scalding nor fermentation reduced the vitamin E con-
of flour was substituted with 7:3 (w/w) mixture of wheat fiber tent. However, the sour dough preparation and dough making re-
and psyllium husk fiber and three antioxidants viz. vitamin C, sulted in 20–60% reduction in the content of tocopherols and to-
β-carotene, and all-rac-α-tocopheryl acetate (ToAc) at levels of cotrienols (Wennermark and Jagerstad, 1992). In the preparation
72, 5.6, and 115 mg. respectively. And pup loaves were made of French bread the content of tocopherols and tocotrienols de-
by the straight-dough procedure. Stabilities of antioxidants in creased during French breadmaking on an industrial scale (Table
bread were studied. The results indicated that the ToAc content 10). Dough making caused a substantial loss of vitamin E. The
was almost stable during storage period of up to 7 days (Park dough making technique of Chorley wood results in a substantial
et al., 1997). A recent American survey found bread to be the incorporation of oxygen in the dough, facilitating the lipoxyge-
fourth highest contributor to vitamin E intake for both men and nase catalyzed oxidation of unesterified polyunsaturated fatty
women (Engelhart et al., 2002). acids. The lipid oxidation and vitamin E destruction were more
A study was undertaken by Leenhardt et al. (2006) to provide pronounced in stored flours than in fresh flours. Whereas in the
solutions to optimize the unsaponifiable antioxidants content of preparation of wheat/rye bread the scalding procedure had no
bread. In the wheat grain, tocopherols are most abundant in the effect on vitamin E retention. Preparation of sourdough from rye
germ and least in the endosperm. There are no significant dif- wholemeal decreased vitamin E content by about 60% vitamin E
ferences for vitamin E activity between the three wheat species activity. This great loss of vitamin E activity was apparent after
(Table 9). Vitamin E losses after bread making were close to 30% 10 min of mixing, suggesting a rapid degradation of vitamin E
without significant differences between the three wheat species. when in contact with water. The low speed mixing used to pre-
There were no varietal differences for vitamin E activity during pare the dough of wheat/rye bread caused lesser losses of vitamin
kneading. Therefore, tocols losses during bread making could E activity of 30% and 18% in the two batches (Niki, 1996).
be attributed to direct oxygenation during dough making and
heat destruction during baking (Leenhardt et al., 2006).
When all-rac-α-tocopheryl acetate (ToAc) was incorporated
into bread formulations, the source of vitamin E it showed good Table 10 Baking of French bread – Effect on vitamin E∗
stability during processing and storage of bread (Cort et al.,
Tocopherol Tocotrienol
1976). In another study conducted by Park et al. (1997) white
bread was fortified individually with fat-coated L-ascorbic acid (mg/100 g dry matter) α β γ α β
AsA, cold-water-dispersible (CWD) β-carotene, and CWD all- French bread
rac-α-tocopheryl acetate (ToAc) at level of 64, 5, and 100 mg Ingredients 0.89 0.41 0.04 0.15 1.65
respectively, of active ingredient per 100 g of flour (14% mois- Dough, freshly mixed 0.60 0.34 <0.01 0.10 1.30
ture basis). When 200 mg of CWD ToAc, which equals 67 mg Dough, after fermented 0.58 0.33 <0.01 0.09 1.24
Bread, freshly baked 0.31 0.17 <0.01 0.05 0.69
of RRR-α-tocopherol equivalents (α-TE), was added to the for-
∗ Reference:
mula of pup loaf, proofed dough retained 96% of added ToAc. (Rogers et al., 1993).
 16 B. NANDITHA AND P. PRABHASANKAR
H3C CH3 CH3
CH3
CH3
Figure 10
Nutritional Importance of α-Tocopherol
It is accepted that the major in vivo function of vitamin E
is as an antioxidant that protects cells and tissues from oxida-
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tive damage induced by free radicals. It may also stimulate im-
mune response, inhibit cancer initiation, lessen the severity of
prostaglandin-mediated disorders, and inhibit the conversion of
nitrites to nitrosoamines, which are strong tumor promoters. It
is known that α-tocopherol scavenges singlet oxygen as well as
oxygen radicals (Niki, 1996).
Brufau et al. (2004) evaluated lipid oxidation after inges-
tion of bakery products enriched with phytosterols, β-carotene
and α-tocopherol. The results of this study agree with the fact
that phytosterols could reduce its absorption of α-tocopherol
(Brufau et al., 2004). The biological activity of various isomers
of vitamin E is listed. Of all the isomers and analogs, RRR-α-
tocopherol, formerly known as d-α-tocopherol, has the highest
biological activity. Tocopherols were used at various levels for
the treatment of diseases.
β-Carotene
β-Carotene (provitamin A) occurs widely in all plants. Car-
rots, green leafy vegetables, and tropical fruits such as papayas
and mangoes are rich sources of β-carotene. β-Carotene is used
mainly as a food colorant. Structurally, β-carotene consists of
two β-ionone rings and an 18-carbon polyene side chain (Fig.
10). β-Carotene and related carotenoids function as singlet oxy-
gen quenchers and free-radical trapping agents. The antioxidant
action is limited to low oxygen partial pressures less than 150
mm Hg, and at higher oxygen pressures β-carotene may be-
come prooxidant. Figure 11 indicates the structure of astaxan-
thin, which is a type of carotenoids.
β-Carotene is insoluble in water and soluble in fats and
oils. Because of the conjugated double-bond structure of the
CH3 CH3 CH3
O
HO CH3
CH3
Figure 11
H3C
CH3 CH3 H3C CH3
β-Carotene.
molecule, β-carotene is sensitive to oxidative decomposition on
exposure to air. On exposure to air, the absorption maximum
drops to 25% of its initial value in 6 weeks at 20◦ C either in
darkness or in light. β-Carotene is also sensitive to light, tem-
perature, and acidic conditions, it is almost completely destroyed
at 45◦ C in air after six weeks. Contact with acids in the pres-
ence of oxygen causes rapid deterioration accelerated by heat.
However, the stability improves on dissolution or suspension in
vegetable oils.
Application in Bakery Products
β-Carotene inhibits the light catalyzed oxidation of fats and
oils such as butter, corn, coconut, rapeseed, groundnut, soybean,
and olive oils.
In an experiment conducted by Park et al. (1997) white bread
was fortified individually with fat-coated L-ascorbic acid (AsA),
cold-water-dispersible (CWD) β-carotene, and CWD all-rac-α-
tocopheryl acetate (ToAc) at level of 64, 5, and 100 mg re-
spectively, of active ingredient per 100 g of flour (14% mois-
ture basis). When 5 mg of β-carotene of the CWD form was
added to the bread formulation, the retention of β-carotene
in proofed dough was 95%. Fresh bread retained 67% of β-
carotene from the CWD form, so the baking loss was 28%. β-
Carotene showed good stability in stored bread. Bread stored
for one to five days retained 67–64% of added β-carotene.
The level of β-carotene decreased to 59% in seven-day-old
bread. Protein Encapsulated (PE) β-carotene, was tested which
is a beaded product in which the β-carotene appears to be en-
trained in a network of fish gelatin. The freeze-dried dough or
bread spiked with PE β-carotene (4 mg/g) showed 101% re-
covery of β-carotene. Proofed dough retained 98% of added
β-carotene. The baking loss with PE β-carotene was 7%, which
was 21% lower than the loss of CWD β-carotene (Park et al.,
1997).
O
H3C OH
CH3 CH3 CH3 H3C CH3
Astaxanthin.
 ANTIOXIDANTS IN BAKERY PRODUCTS
Quilez et al. (2006) reported a sensory evaluation of
croissants and magdalenas (Spanish muffins) has been carried
out to compare them with the same products with phytosterol
ester, α-tocopherol, and β-carotene added. On the total of 30
replies, it was observed that the majority response in the two
groups was that differences were not found between samples.
His study indicated that the phytosterol esters, α-tocopherol
and β-carotene, are particularly suitable for inclusion in
bakery products, as functional ingredients, since the addition
of effective doses does not affect the appearance or taste.
Furthermore, the large differences in the composition of the
commercial croissants and muffins and therefore, in their
nutritional value, makes the use of these and other functional
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ingredients recommendable in muffins, sponge cakes, and in
general in all the bakery products (Quilez et al., 2006).
Retention of β-Carotene after Processing
The study was undertaken to examine that stability of β-
carotene by Rogers et al. (1993). Three bakery products, yellow
cake, sugar cookies, and bagels, were prepared with β-carotene
added to the formula shortening. Products were sampled at vari-
ous intervals during processing and during storage. For all prod-
ucts, prebaking processing steps had no adverse effect on the
stability or isomeric distribution of carotene. Carotene losses
during baking ranged from 20% in bagels and cake to 30% in
cookies. During baking, the all-tans isomers were reduced from
91% in the spiked shortening to 85, 77, and 74% in bagels, cake
and cookies respectively. A serving of each product provided
about 1 mg of β-carotene (Rogers et al., 1993). Bauernfeind
et al. (1958) reported 74–95% retention of total β-carotene in
cookies, piecrust, and yellow cakes (Bauernfeind et al., 1958).
Whole wheat bread and crackers were fortified with β-
carotene using three pure (all-tans) sources: one water-soluble
and two oil-soluble. This study was focused on two whole-
wheat products (bread and crackers) and assessed the stabil-
ity characteristics of β-carotene added to those products. How-
ever, carotene losses were appreciable during baking, ranging
from 4–15% for bread products to 18–23% for crackers. β-
Carotene sources with added antioxidants generally exhibited
greater carotene stability, but only during baking and not during
product storage. Baking caused a steric shift in carotene, with cis
isomers averaging 18% in bread products and 34% in crackers
(Ranhotra et al., 1995).
The investigation of Park et al. (1997) involves fortification
of bread with a combination of three antioxidants viz. vitamin
C, β-carotene and all-rac-α-tocopheryl acetate. The retention
of β-carotene was excellent in the fresh bread, but a 40% loss
occurred after seven days of storage (Park et al., 1997).
Compared to Vitamin E, carotenoids suffer greater loss due
to wheat lipoxygenase activity during breadmaking as reported
by Leenhardt et al. (2006). Carotenoid pigments have been de-
termined after each step of bread making.
Measurements were performed on the initial flour, on the
dough after kneading, and on freshly baked bread. Total
17
Table 11 Total carotenoids contents before (whole wheat flour) and after
breadmaking (whole-wheat breads), expressed in μg/g of dry weight∗
Carotenoids (μg/g dw)
T. monococcum
Whole flour 5.75
Whole bread 3.34
T. durum
Whole flour 3.32
Whole bread 1.61
T. aestivum
Whole flour 1.24
Whole bread 0.10
∗ Reference: (Niki, 1996).
carotenoids content, calculated by summing lutein and zeax-
anthin concentrations, decreased after breadmaking (Table 11).
The carotenoids content of bread wheat decreased by about 66%
after kneading, probably because of the presence in wheat flour
of enzymes such as lipoxygenase (LOX) and peroxidase (POD),
which became active when, water was added. The percentages
of pigment losses were highly correlated to LOX activity of
the respective flours. As a result, carotenoids oxidation during
kneading was assumed to depend mainly on LOX activity.
Carotenoids Enhance Vitamin E Antioxidant Efficiency
The principal role of vitamin E is that of a free radical scav-
enging antioxidant, and there is evidence that high intake of
carotenoids (particularly β-carotene) and α-tocopherol may pre-
vent cancer and other diseases. Bohm et al. (1997) reported
that the results of pulse radiolysis and laser flash photolysis
studies of the reactions of the α-tocopheroxyl radical with a
range of carotenoids and of the interaction of ascorbic acid with
the carotenoids radical cations. It has been proposed that α-
tocopherol protects β-carotene by a repair mechanism.
The rate constants reported in the above study are not
measured in a biological environment, but the high values
obtained indicated the direction of electron transfer, from
carotene to α-tocopherol radical, and therefore the repair of
the α-tocopheroxyl radical by the carotenoids. Hence, all the
carotenoids studied are capable of regenerating α-tocopherol
from its radical. Astaxanthin does not recycle α-tocopherol, but
it may be recycled by α-tocopherol.
A synergistic effect observed in cell protection by β-carotene
and vitamins E and C may be related to the fact that β-carotene
is not only quenching oxy-radicals but is also repairing the
α-tocopheroxyl radical, which is produced when α-tocopherol
scavenges an oxy-radical. A synergistic effect may reflect dif-
fering cellular locations of β-carotene and α-tocopherol. It is
generally accepted that β-carotene is more lipophilic than α-
tocopherol and will be more likely to be in the interior of a
membrane. However, the β-carotene radical is a charged species,
while the α-tocopherol radical is uncharged (Bohm et al., 1997).
 18 B. NANDITHA AND P. PRABHASANKAR
H OH
H
CH3
O 1997).
O H H Park et al. (1997) fortified white bread with fat-coated L-
HO OH
Figure 12 Ascorbic acid.
Ascorbic Acid (Vitamin C)
Ascorbic acid is ubiquitous. For the vast majority of mam-
mals, ascorbic acid is not a vitamin because their liver enzymes
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have the ability to convert glucose to ascorbic acid. We are
dependent on adequate intake of ascorbic acid for prevention
of scurvy and for overall well being. It was not known until
1922 when Albert Szent-Gyorgyi isolated a white crystalline
substance from the adrenal cortex of cows, which he first be-
lieved to be a new adrenal hormone. He named this substance
hexuronic acid and went on to demonstrate that this substance
was vitamin C. He later discovered that Hungarian paprika was
a much better source of this vitamin and renamed the substance
ascorbic acid because it prevented scurvy.
Ascorbate has unusual structure in that it has at its chemical
core a five-membered lactone ring: on this ring is a bifunc-
tional ene-diol group with an adjacent carbonyl group (Fig. 12).
This conjugated structure gives ascorbate a very rich chemistry.
Ascorbate is an excellent reducing agent.
Pure ascorbic acid solutions are colorless. Ascorbate is ex-
tremely soluble in water, ≈ 1 g dissolves in 3 ml water. It is in-
soluble in nonpolar organic solvents such as benzene, petroleum
ether, fats, and their solvents (Buettner and Jurkiewicz, 1996). It
is the least stable vitamin in the food supply. Crystalline ascorbic
acid (AsA) is added to foods to restore AsA losses (Bauernfeind,
1985).
Application in Bakery Products
Ascorbic acid is used as an antioxidant in many food prod-
ucts, including processed fruits, vegetables, meat, fish, dairy
products, soft drinks, and beverages. Ascorbic acid (AsA) func-
tions synergistically with phenolic antioxidants such as BHA,
PG and tocopherols in retarding oxidation in fats and oils. Ad-
dition of AsA as a nutrient in white pan bread has not been
successful. More than 50% of AsA was lost in the process of
making fortified bread, and almost no AsA survived after stor-
age of bread at 25◦ C for 7 days (Hung et al., 1987). Physical
and chemical attributes and consumer acceptance of sugar-snap
cookies containing naturally occurring antioxidants were stud-
ied by Hix et al. (1997). The antioxidant compounds, ascorbate
(ascorbyl-2-phosphate), α-tocopherol, a combination of ascor-
bate and α-tocopherol were tested as replacement for BHA in
preserving sugar-snap cookies. The cookies with various antiox-
idants were compared for their shelf life, moisture content, and
width, stacking height, surface score, color, and texture. The
results obtained indicated that natural antioxidants could be ef-
fectively substituted for BHA in sugar-snap cookies (Hix et al.,
ascorbic acid at a level of 64 mg of active ingredient per 100
g of flour (14% moisture basis). Ascorbic acid was assayed
by reverse-phase high-performance liquid chromatography (RP-
HPLC) with electrochemical detection. L-Ascorbic acid is often
added at low level (<100 ppm) to bread dough to modify its
rheological properties. But at those levels, practically no AsA
remains in freshly baked bread. The AsA retention in proofed
dough, immediately before baking was 99%. During the baking
step, ≈23% of AsA was destroyed, and much more degraded
during seven days of bread storage at room temperature. When
AsA was added to bread dough in combination with β-carotene,
the retention of β-carotene increased significantly from 95 to
101% in proofed dough (Park et al., 1997).
Retention of Ascorbic Acid after Processing
Park et al. (1997) conducted an experiment by incorporat-
ing 10% weight of flour with 7:3 (w/w) mixture of wheat fiber
and psyllium husk fiber and fortified with a combination of vi-
tamin C, β-carotene, and all-rac-α-tocopheryl acetate (ToAc)
at levels of 72, 5.6, and 115 mg respectively and the stabilities
of antioxidants were studied. The AsA retention in the proofed
dough, immediately before baking, was 98% for the fiber and
antioxidant dough. Over a seven-day storage period, AsA also
disappeared significantly faster in the fiber and antioxidant bread
than in the control bread (Park et al., 1997).
L-Ascorbic acid and its 2-phosphorylated derivatives are for-
tified into bread and its antioxidant properties were studied by
Wang et al. (1995) L-Ascorbate 2-mono- and 2-polyphosphates
(AsMP and AsPP) has high vitamin C activity and high stability
toward oxygen. After 6 days storage at 25◦ C, 32–45% of vitamin
C was retained when AsPP and AsMP had been added at 400
and 560 mg AsA equivalents per pup-loaf, respectively, only
17% remained when 560 mg AsA had been added. When high
level of vitamin C was added, ≈40% was lost from fresh bread
made with AsMP. When AsPP was mixed into bread dough at
400 mg equivalents of AsA, breadmaking losses of vitamin C
were 35% in the presence of reduced iron in the dough vs 43%
in the presence of ferrous iron. Bread enriched with ferrous iron
and fortified with 560 AsA equivalents/100 g flour in the form
of AsPP and AsA retained 40% and 5% vitamin C respectively,
after 6 days at 25◦ C (Wang et al., 1995).
Pup-loaves of white pan bread were fortified with three com-
mercial forms of fat-coated AsA and with uncoated AsA at a
level equivalent to 64 mg of sample per 100g of flour on 14%
moisture basis by Park et al. (1994). Sample 1 contained large
crystals of AsA coated with 30% fat (mp 58–71◦ C) and resisted
leaching in 6% metaphosphoric acid at 25◦ C. Sample 3 con-
tained mostly small crystals coated with 10% fat (mp 49–69◦ C)
and lost 70% of its AsA during leaching. Proofed bread dough
mixed with sample 1 retained 80% of fat-coated AsA, whereas
 ANTIOXIDANTS IN BAKERY PRODUCTS
proofed bread dough mixed with sample 3 retained only 7%
of fat-coated AsA. Bread fortified with sample 1 showed 20%
higher retention of vitamin C than bread fortified with sample
3 or with uncoated AsA. Bread fortified with sample 3 (hy-
drogenated soy bean oil and 70% AsA) showed intermediate
retention of vitamin C. Stability of vitamin C during baking and
storage of pup-loaves fortified with uncoated and fat-coated AsA
was studied and showed that the retention of AsA in pup-loaves
containing the fat-coated samples was related inversely to the
level of free AsA in the proofed doughs. Thus, the fat coating on
AsA inhibited the destruction of AsA during baking. Stability of
vitamin C in bread fortified with AsA, AsMP and AsPP shows
the retention of total AsA equivalents during room-temperature
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storage of bagged pup-loaves fortified with vitamin C in the
forms of AsMP and AsPP. Hydrolysis of AsPP and AsMP to
AsA in bread dough was studied and the results of adding AsMP
to bread dough were similar to those of adding AsPP, except that
the quantities of AsMP retained after mixing, proofing and bak-
ing were 7, 6, and 3% less, respectively (Park et al., 1994).
Natural Antioxidants from Wheat
Wheat is a major crop and an important component of the hu-
man diet, particularly in developing countries. The antioxidant
activity of wheat kernel is very low. Wheat grain had minimal
antioxidant activity while the bran fraction had low antioxidant
activity. Ferulic acid and p-coumaric acid were predominant
phenolic acids in hard red spring (HRS) wheat hulls while fer-
ulic was the predominant phenolic acid in HRS wheat and flour.
Durum wheat varieties protected oil oxidation equally well and
were found to contain similar phenolic acid profiles. The wheat
bran extract was composed of protocatechuic acid (226 ppm), p-
hydroxybenzoic acid (124 ppm), gentisic acid (108 ppm), caffeic
acid (116 ppm), vanillic acid (637 ppm), cholorogenic acid (84
ppm), syringic acid (130 ppm), p-coumaric acid (580 ppm), and
ferulic acid (764 ppm). The durum wheat extract was a signifi-
cantly better antioxidant than the individual phenolic acids ex-
cept protocatechuic acid and cholorogenic acid (Hall III, 1996).
Wheat is an important dietary source of vitamin E. During
manufacturing of commercial wheat products for human con-
sumption, a marked reduction in vitamin E content occurs. In
the production of white wheat flour from whole grain wheat, the
vitamin E content is reduced by about 50% due to the removal
of bran and germ. The susceptibility of vitamin E to oxidation is
another important cause of losses during processing and storage.
Antioxidant Activity of Wheat
Antioxidant activity of wheat grain, flour, and its fractions and
retention of antioxidant activity were studied by various scien-
tists. Distribution of wheat carotenoids in milling streams was
studied by Leenhardt et al. (2006) The amounts of carotenoid
pigments vary in the different milling fraction according to their
repartition in the kernel. Carotenoids were present in all milling
19
fractions, but the high concentration observed in the flour points
to a higher concentration in the wheat endosperm than in other
parts of the grain (Leenhardt et al., 2006).
Storage stability of wholemeal flour, white flour, brand germ
of wheat were investigated by Wennermark et al. (1992). The vi-
tamin E activity of the various wheat fractions was retained to al-
most the same extent, wholemeal (60%), white flour (62%), bran
(72%), and germ (60%), after 12 months of storage when stored
at 20◦ C. Pure wheat germ contains almost exclusively α- and β-
tocopherol. When comparing the retentions of the tocopherols
and tocotrienols present in wheat after 12 months of storage
at 20◦ C, α-tocopherol and α–tocotrienol showed lower reten-
tions than the corresponding β-isomers. There were no losses
of any tocopherols or tocotrienols in four wheat fractions when
stored at –40◦ C for 12 months. Storage at 20◦ C for 12 months
decreased vitamin E activity of the various wheat fractions by
28–40% (Wennermark and Jagerstad, 1992).
The effect of milling on the phenolic content and antioxi-
dant capacity of two wheat cultivars, namely WAD (Canadian
Western Amber Durum: Triticum turgidum L. var. durum) and
CWRS (Canadian Western hard Red Spring: Triticum Aestivum
L.) were studied by Chandrika et al. (2007) During milling
of wheat several fractions were obtained namely bran, flour,
shorts, and feed flour. In addition, semolina was the end product
of durum wheat milling. Among different milling fractions the
bran had the highest phenolic content while the endosperm pos-
sessed the lowest amount and this was also reflected in reducing
power and iron chelation capacity of different milling fractions
in the two cultivars. This study demonstrated the importance of
bran in the antioxidant activity of wheat; hence the consump-
tion of whole-wheat grain may render beneficial health effects
(Chandrika et al., 2007).
The objectives of the study conducted by Adom et al. (2003)
were to determine the phytochemical profiles and total antiox-
idant activity for 11 diverse wheat varieties and experimental
lines. Flavonoid contents of wheat varieties tested are expressed
as micromoles of catechin of equivalent per 100 g of grain.
W7985 had the highest free Flavonoid content among all of the
varieties tested. The total antioxidant activities of 11 wheat va-
rieties were studied. Bound phytochemical of wheat contributed
the majority of total antioxidant activity of wheat extracts. Bound
phytochemicals contributed >82% of total antioxidant activity
and free forms contributed 12–35% (Adom et al., 2003).
Three hard winter wheat varieties (Akron, Trego, and Platte)
were examined and compared for their free radical scaveng-
ing properties and total phenolic contents (TPC). Free radical
scavenging properties of wheat grain extracts were evaluated by
spectrophotometric and electron spin resonance (ESR) method
by Yu et al. (2002). Wheat extracts were measured and com-
pared for their free radical scavenging activities against rad-
ical cation ABTS•+ (2,2 -Azino-di[3-ethylbenzthiazoline sul-
fonate]). Platte wheat had the greatest activity to quench
ABTS•+ , followed by Akron and Trego. ESR spectra showed
that extracts from all three wheat varieties directly reacted and
quenched free DPPH (2,2-diphenyl-1-picryhydrazyl) radicals at
 20 B. NANDITHA AND P. PRABHASANKAR
the tested concentration. The 60 min reaction time resulted in
a greater reduction in free radicals than the 10 min reaction
time. Trego grain had the lowest activity against both DPPH
and BTS•+ radicals. The relative radical quenching activities of
Akron, Platte and Trego grains are 16.5:8.1:1 against DPPH (Yu
et al., 2002).
Extracts from three winter wheat varieties (Trego, Akron, and
Platte) were evaluated and compared to α-tocopherol for their
inhibitory effects on lipid peroxidation in fish oils by measur-
ing the oil stability index (OSI) by Yu et al. (2002). The OSI
represents a measure of the resistance of lipid to oxidation, with
OSI duration being positively associated with oil or food sta-
bility. The Rancimat is an instrument that automates the OSI
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measurements. The extracts from all three hard winter wheat
varieties showed inhibitory effects against lipid peroxidation in
fish oil. This inhibition is about 3.1 times stronger than that
provided by 300 ppm α-tocopherol. Trego extracts may have
potential to be developed as food antioxidant for commercial
application. The order of the radical scavenging activity against
DPPH radical is Akron extracts (AS) > Platte extracts (PS)
> Trego extracts (TS), under the experimental conditions. The
greatest chelating capacity was detected in Akron extracts (5.44
mg/g extracts), but not Trego (5.41 mg/g extracts) (Yu et al.,
2002).
Miller et al. (2000) described the significance of whole grains
products and antioxidants. Whole grain bread had 2000 TE
(Trolox equivalents) compared to white bread at 1200 TE, indi-
cating the contribution of bran and germ. Whole grain wheat and
oat based products average higher in antioxidants than products
from refined rice or corn ingredients (Miller et al., 2000).
Four cereals including barley, pearl millet, rye, and sorghum
were evaluated in terms of their composition of dietary
fiber, resistant starch, minerals, and total phenols and an-
tioxidant properties by Ragaee et al. (2006). Antioxidant ac-
tivity was evaluated on the basis of scavenging capacity of
2,2-diphenyl-1-picrylhydrazyl (DPPH• ) radicals and 2,2 -azino-
di-[3-ethylbenzthiazoline sulphonate] ABTS•+ radical cations.
Whole grains and wheat flours significantly differed in total
phenol content ranging from 501 to 562 μg/g in wheat flours
and from 879 to 4128 μg/g in whole grains. The ABTS test
is based on the formation of ABTS•+ by reacting ABTS with
Table 12 Chemoprotective antioxidants from fruits and vegetables∗
Source Active component
Apples, Strawberries Vitamin C, bioflavonoids, chalcones
Leafy greens, cabbage, broccoli, Vitamin C, lutein, zeaxanthin
cauliflower
Carrots α- and β-carotene, phenolic compounds
Grapes, red wine Phenols, catechins
Citrus fruits β-Cryptoxanthin, bioflavonoids, chalcones, vitamin C
Garlic, onion, leeks Allicin, flavonoids, vitamin C, selenium, sulfur
∗ Reference: (Madhavi et al., 1996).
metmyoglobin and H2 O2 at 37◦ C. The ABTS•+ has a rela-
tively stable blue-green color which is measured at 600 nm.
In the presence of an antioxidant such as 6-hydroxy-2, 5,7,8-
tetramehylchroman-2-carboxylic acid (trolox) or potential an-
tioxidants in material extracts, the color production will be sup-
pressed to a certain extent in proportional to the concentration
of antioxidants. The antioxidant properties of the three grains
were comparable to butylated hydroxytoluene (Ragaee et al.,
2006).
Antioxidants from Fruits
Epidemiological studies indicate that frequent consumption
of fruits is associated with lower risk of stroke and cancer. Fruit
Dietary Fiber (DF) concentrates have, in general, better nutri-
tional quality than those found in cereals, because of their sig-
nificant contents of associated bioactive compounds (flavonoids,
carotenoids, etc.) and more balanced compositions. According
to recent reports, the presence of significant amounts of bioac-
tive compounds, such as flavonoids and carotenoids, in DF from
fruits imparts them considerable nutritional value. Some of the
chemoprotective antioxidants from fruits are indicated in Ta-
ble 12. The antioxidant properties of flavonoids and carotenoids
come from their ability to link free radicals that easily attack
saturated fatty acids present in cell membranes, causing perox-
idation, decreasing permeation, and damage of membrane pro-
teins, leading to cellular inactivation. DNA is also subject to free
radical effects producing mutations, which may lead to cancer.
Antioxidative properties of several fruits and their application
in bakery products have been studied. Some of them are given
below.
Mango Fruit
Vergara-Valencia et al. (2007) characterized a mango dietary
fiber concentrate (MDF) with antioxidant capacity, using the
unripe fruit. MDF was obtained and its extractable polyphenols
and anti radical efficiency was evaluated. Extractable polyphe-
nols or total soluble polyphenols (EPP) content in MDF was
16.1 mg/g. Both types of bakery products (cookie with MDF
Mechanism of action
Antioxidant
Scavenger of ROS, antioxidant, suppresses
promotion of lung tumors in mice
Antioxidant, pS2 gene expression, inhibits tumors
in rats and mice
Antioxidant
Antioxidant, stimulates expression of RB gene and
p73 gene
Detoxifies carcinogen, inhibits Helicobacter pylori
 ANTIOXIDANTS IN BAKERY PRODUCTS 21

Table 13 EPP and AE of bakery products added with MDF∗ OH

Bakery product

Cookie Control Bread Control HO O OH

Component with MDF cookie with MDF bread

EPP 11.8 +/–1.2 Not done 10.1 +/–0.5 Not detected OH

OH
EA 9.1 +/–0.6 Not detected 8.6 +/–0.3 Not detected
∗ Reference:
OH
(Reddy et al., 2005).
Figure 13 Gallocatechin.
and bread with MDF) containing MDF exhibited similar levels
of EPPs (Table 13). The results suggested that baking does not Antioxidant DF can be obtained from red and white whole grape
have significant impact on these compounds. The products with pomaces originated from wine or grape juice production, as
MDF have significant antioxidant properties (Vergara-Valencia well as from white and red grape skins and seeds. Usual ranges
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et al., 2007). of ADF composition in these products are DF (50–75%), EPP

Guava Fruit
Guava (Psidium guajava) is an important tropical fruit,
mostly consumed fresh. The fruit is a berry, which consists
of a fleshy pericarp and seed cavity with flesh pulp and nu-
merous small seeds. Pulp and peel fractions were tested, and
both showed high content of dietary fiber (48.55–49.42) and ex-
tractable polyphenols (2.62–7.79%). The antioxidant activity of
polyphenols compounds was studied, using three complemen-
tary methods by Jimenez-Escrig et al. (2001). A 1 g (dry matter)
portion of peel contained DPPH• activity; FRAP activity, and
inhibition of copper-induced in vitro LDL oxidation, equivalent
to 43 mg, 116 mg, and 176 mg of Trolox, respectively. The re-
sults indicated that guava could be a suitable source of natural
antioxidants (Jimenez-Escrig et al., 2001).
Banana Fruit
Bananas are one of the most popular fruits in the world
market. The antioxidant compounds from commercial bananas,
Musa Cavendish, were studied by Someya et al. (2002). Gallo-
catechin was more abundant in peel (158 mg/100 g dry weight)
than in pulp (29.6) mg/100 g dry weight). Total phenolic were
more abundant in peel (907 mg/100 g dry weight) than in pulp
(232 mg/100 g dry weight). The peel extract showed 2.2 times
stronger antioxidant activity than the pulp extract when the incu-
bation times were compared. One of the antioxidant compounds
in the bananas was determined to be gallocatechin, which was
related to the antioxidant activity of the banana extract. The
structure of gallocatechin is shown in Fig. 13. Hence bananas
should be considered as a good source of natural antioxidants
for foods (Someya et al., 2002).
(extractable polypheonols) (1–9%) and NEPP (non-extractable
polyphenols) (15–30%). The antioxidant capacity of this prod-
uct was in vitro determined by lipid oxidation inhibition and free
radical scavenging procedures. One gram of the product showed
similar effect than 400 mg and 100 mg of DL-α-tocopherol, re-
spectively (Sauro-Calixto, 1998).
Amla and Raisins
Reddy et al. (2005) used three-plant foods viz., amla (Emblica
officianalis), drumstick leaves (Moringa oleifera) and raisins
(Vitis vinifera) as sources of natural antioxidants. All the three
extracts exhibited a high percentage of antioxidant activity eval-
uated using β-carotene-linoleic acid in vitro system, compared
to synthetic antioxidants. Biscuits prepared by addition of nat-
ural extracts were subjected to sensory studies and chemical
analysis. Lipids extracted from biscuits were analyzed for the
stability during storage (Table 14). Biscuits treated with natural
antioxidants, extracted from raisins (B4) and drumstick leaves
(B5) received higher panel scores during storage period of 6
weeks, than control (B1), BHA (B2), and amla (B3) extract
incorporated biscuits. Extracts from drumstick leaves and amla
were more effective in controlling lipid oxidation during storage
(Reddy et al., 2005).
Antioxidants from Spices
Recently, plants have received much attention as sources of
biologically active substances including antioxidants, antimu-
tagens, and anticarcinogens. Additions of freeze-dried extracts
from fenugreek seeds and ginger rhizomes to beef patties are
Table 14 Antioxidant activity (%) in biscuit lipids∗

Biscuit sample Initial 2nd week 4th week 6th week

Grape Fruit
B1 30 13 13 9
The main characteristics of a natural product, antioxidant B2 70 68 62 60
dietary fiber (ADF), rich in both dietary fiber (DF) and polyphe- B3 73 62 52 50
nolic compounds (PP), obtained from red grape pomace is de- B4 57 50 45 36
B5 95 89 78 68
scribed by Sauro-Calixto (1998). Polyphenolic compounds in
∗ Reference:
vegetables are found free or bound to cell walls or to protein. (Borrelli et al., 2003).
 22 B. NANDITHA AND P. PRABHASANKAR
reported to be effective in controlling lipid oxidation during cold
storage. Natural aromatic plants and spices have been widely
used in many food products such as meat and meat product, dairy,
and bakery products. The keeping quality of baked foods such as
crackers, cookies, and biscuits is of great economic importance
since these products are widely used and are often stored for
extended periods before consumption. The spices most com-
monly used in bakery products are cinnamon, mint, nutmeg,
mace, cloves, poppy and sesame seeds.
Bassiouny et al. (1990) studied the effects of some plant ma-
terials and their extracts on the stability of fat in soda cracker bis-
cuits. A soda cracker biscuit was processed using a fine powder
of marjoram, spearmint, peppermint, and basil and their purified
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ether extracts as natural antioxidants. Addition of purified ether
extract of each of the four plant materials gave an excellent an-
tioxidant effect on the biscuit compared with the effect of BHA
at concentrations of 0.01%, 0.02%, and 0.03%. Addition of a
fine powder of all plant materials at 0.5% gave an antioxidant
effect on the biscuit, compared to the control sample. Addition of
a 1% mixture of equal amounts of the four plant powders caused
a pro-oxidant effect in the biscuit (Bassiouny et al., 1990).
Curcumin (diferuloyl methane), the natural yellow pigment
in turmeric, is isolated from the rhizomes of the plant Curcuma
longa by Joe et al. (2004). The discovery of the antioxidant
properties of curcumin explains many of its wide-ranging phar-
macological activities. Curcumin is an effective antioxidant and
scavenges superoxide radicals, hydrogen peroxide, and nitric
oxide from activated macrophages. Lipid peroxidation is lower
in liver, kidney, spleen, and brain microsomes from retinal de-
ficient rats that are fed with 0.1% dietary Curcumin for three
weeks. Curcumin has beneficial effects and certainly qualifies
for serious consideration as a pharmaceutical or nutraceuticals
or phytoceutical agent (Joe et al., 2004).
Mint is the third largest popular flavor worldwide. It appeals
to all sections of people irrespective of age, gender, and ethnic
background. It is regarded as one of the most important spices
throughout the world. Mint belongs to a small genus of aromatic
perennial herbs distributed mainly in the temperate regions of the
world. Mint is incorporated in biscuits in various forms (powder,
extract, and menthol crystals) at different levels as an antioxi-
dant. The effect of incorporation on the quality characteristics
(texture and color) and acceptability of biscuits was also studied
during the storage period by Bajaj et al. (2006). The texture of
biscuits with mint powder (MNT-P) was comparable with the
control (CNT) and BHA (BNT) biscuit indicating their crisp
nature. The study indicates that MNT-P biscuits were highly ac-
ceptable compared to mint extract (MINT-E) and pure methanol
(MNT-M) biscuits (Bajaj et al., 2006).
Maillard Reaction Products as Antioxidants
Maillard reaction products formed during heat processing
or storage of food products are known to have significant an-
tioxidant properties. Melanoidins, the brown colored polymers
formed through Maillard type reaction in several heat-treated
foods, represents a significant part of our diet. The reaction
between carbohydrates and protein produce the Maillard reac-
tion products (MRPs) responsible for the brown color and the
organoleptic properties of bread and bakery products.
An experiment was conducted on characterization of colored
compounds obtained by enzymatic extraction of bakery prod-
ucts by Borrelli et al. (2003). The antioxidant activity of the
different fractions obtained by the gluten-glucose model system
digested with Pronase has been measured by the ABTS method.
The high molecular weight fraction (above 10,000 Da) has the
highest antioxidant activity. The main component responsible
for the antioxidant activity is a pyrrolinone reductones named
PRONYL-glycine (Borrelli et al., 2003).
The generation of a chain-breaking antioxidant capacity has
been analyzed in butter cookies, as a function of the cooking
time and thus, of the browning related to the formation of MRP
(Maillard reaction products). Results indicated that during the
first 20–30 min of cooking, when browning takes place, an an-
tioxidant capacity accounting for up to 5 g of Trolox is produced
in 100 g of dried aqueous extracts of the cookies. This result
supports the concept that functionally relevant antioxidants are
generated by Maillard reaction (Bressa et al., 1996).
Stability of antioxidants formed from histidine and glucose
by the Maillard reaction was studied by Lingnert and Waller
(1983). The antioxidants were found to be unstable in solu-
tion when exposed to air. When they were stored in nitro-
gen atmosphere, no loss of antioxidative effect was noticed.
The oxygen sensitivity was found to be pH dependent. In the
dry stage and in concentrated solutions at low temperatures,
the MRP were found to be fairly stable (Lingnert and Waller,
1983).
Antioxidants from Other Sources
Apart from the common sources of antioxidants such as
spices, fruits, and vegetable, antioxidants are also found in many
other sources such as milk, cocoa, seaweeds, vanilla, garcinia,
agro-industrial wastes, and etc., and these antioxidants still have
not found any application in bakery products. Their antioxidant
property is studied in isolation. Some of them are used in edible
oils.
Milk
Milk contains several antioxidant factors, like vitamins and
enzymes. Possible antioxidant activity of milk proteins and hy-
drolysates has also been shown. Milk-derived antioxidative pep-
tides are composed of 5–11 amino acids including hydrophobic
amino acids, proline, histidine, tyrosine, or tryptophan in the
sequence. Caseins have been shown to prove antioxidant activ-
ity against TBARS in both Fe/ascorbate induced peroxidation
of arachidonic-derived liposomes and model linoleic acid sys-
tems. The peptide, Tyrosine-Phenyl alanine-Tyrosine-Proline-
 ANTIOXIDANTS IN BAKERY PRODUCTS
Gluten-Leucine, was found to possess a potent superoxide an-
ion radical scavenging activity. Caseins have polar domains that
contain phosphorylated serine residues, and their characteristic
sequences, -SerP-SerP-SerP-Glu-Glu, are effective cation chela-
tors that form complexes with calcium, iron, and zinc. Casein
hydrolysates and low molecular weight casein hydrolysates had
better peroxyl radical scavenging activities than enriched CPP.
In β-lactoglobulin, one of the peptide, Trp-tyr-Ser-Leu-Ala-Ala-
Ser-Asp-Ile has possessed higher radical scavenging activity
than butylated hydroxyanisole (BHA) (Pihlanto, 2006).
Vanilla
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Vanilla has been known in Europe since the beginning of
the sixteenth century when it was introduced into Spain from
South America. An experiment was conducted to separate the
two function of vanillin, and to show that potent antioxidant
properties of vanillin increased the shelf life of precooked dried
cereals. A preliminary examination of the results showed that
the addition of vanillin during the processing had only a small
influence on retarding the development of oxidative degradation
in dried cereal flakes. The formation of pentane was used as an
indicator for the specific oxidative degradation of linoleic acid.
The wet mix series of rice flakes shows about a 50% decreased
pentane formation for 0.5% vanillin, compared with the blank
after 4 months storage (Burri et al., 1989).
Cocoa (Theobroma cacao L.)
Cocoa shoot (CS), young leaves (CL), and tea leaves (GT)
were processed according to green tea processing procedures.
Polyphenolic components were extracted and analyzed using
high-pressure liquid chromatography by Osman et al. (2004).
The total phenolic content is significantly higher in CL (28.4%),
than in CS (19.0%) and GT (17.3%). The main catechin polyphe-
nols in extracts were epicatechin (EC), epigallocatechin gallate
(EGCG), epigallocatechin (EGC), gallic acid (GA), and epi-
catechin gallate (ECG). The polyphenol extracts (CS and CL)
showed similar antioxidation powers to GT and BHA through-
out the entire concentrations range (100–2000 ppm). In the oil-
based test medium the antioxidative performance of polyphe-
nol extracts were better than BHA at 50 ppm (Osman et al.,
2004).
Seaweeds
Evaluation of antioxidant activity of several edible seaweeds
was done by Jimenez-Escrig et al. (2001). The seaweeds used
and evaluated are Fucus vesiculosus L., Laminaria ochroleuca
L., Undaria pinnatifida Harvey, Chondrus crispus Stackh, and
Porphyra umbilicalis. The order of scavenging activity for the
tested seaweeds was Fucus > Laminaria > Undaria > Por-
phyra. Ferric-reducing antioxidant power of Fucus was more
than Porphyra and Laminaria. The antioxidant activity and the
content of phenolic compounds decreased with processing and
23
storage in the seaweeds tested. In addition, Fucus showed good
efficiencies in the in vitro inhibition of LDL oxidation (Jimenez-
Escrig et al., 2001).
Garcinia Extracts
Garcinia is a large genus of polygamous trees or shrubs,
distributed in tropical Asia, Africa, and Polynesia. The fruits
of Garnicia are not palatable due to their acid taste. Garnicia
is a rich source of secondary metabolites including xanthones,
flavonoids, benzophenones, lactones, and phenolic acids. In
a study conducted, the hexane and chloroform extracts from
G. cowa and G. pedunculata were studied for their antioxidant
capacity by the formation of phosphomolybdenum complex at
100 ppm concentration and reducing power by potassium fer-
ricyanide reduction method at various concentrations. Hexane
and chloroform extracts from G. cowa showed higher antioxi-
dant capacity than G. pedunculata extracts. Similarly, both the
extracts from G. cowa showed higher reducing power than the
extracts from G. pedunculata (Joseph et al., 2005).
In another study conducted by Jayaprakasha et al. (2006) hex-
ane and chloroform extracts from the fruit rinds of Garcinia pe-
dunculata were tested for their antioxidative and antimutagenic
activities. Both the hexane and chloroform extracts showed an-
tioxidant activity studied through β-carotene-linoleate model
system and α, α-diphenyl-β-picrylhadrazyl (DPPH) method at
various concentrations. At 500 ppm concentration, in case of β-
carotene-linoleate model system, the hexane and chloroform ex-
tracts of G. pedunculata showed 60% and 67% antioxidant activ-
ity respectively, whereas the free radical scavenging activity was
45% and 65% respectively with DPPH method (Jayaprakasha
et al., 2006).
Agro-Industrial Wastes
The normal antioxidants from residual sources may be used
for increasing the shelf life of food by preventing lipid peroxida-
tion and protecting from oxidative damage (Moure et al., 2001).
Antioxidant activity of some of the agro-industrial wastes are
listed in Table 15.
Influence of Baking Processing Conditions on the Overall
Antioxidant Properties
Preservation methods are generally believed to be responsi-
ble for a depletion of naturally occurring antioxidants in bakery
products (Nicoli et al., 1999). There are several possible effects
of baking on the overall antioxidant potential of baked products.
There may be formation of novel compounds having antiox-
idant activity: Maillard reaction products (MRPs), which can
be formed as a consequence of intense heat treatment or pro-
longed storage, generally exhibit strong antioxidant properties.
Changes in antioxidants during baking are represented in Ta-
ble 16. There may be formation of novel compounds having
 24 B. NANDITHA AND P. PRABHASANKAR

Table 15 Antioxidant activities of extracts from residual materials from agro-industrial origin∗

Residue Antioxidant activity assay Activity (conc. Antioxidant)

Durum wheat bran Soy oil oxidation PV (0.05%), 37.6–42 meq/kg

PV (0.05% BHA–BHT), 22.0 meq/kg
PV (control), 129.0 meq/kg
Peanut hulls DPPH radical scavenging IP (extract 1.5 mg/ml), 89.3%
IP ∗ BHA 240 μM), 92.6%
IP (catechin 8 μM), 89.3%
Grape seed Lecithin liposome oxidation ILLO (0.1 mM), 86%
ILLO (0.1 mM BHT), 88.5%
ILLO (+) catechin, 40%
Grape pomace Cu-induced human LDL oxidation NPIT (catechin 3 μM GA eq), 110.4 min
Grape seed extract Antiulcer activity (200 mg/kg) Lesion length (control), 111 mm
Lesion length (catechin), 88 mm
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Lesion length (extract), 4–20 mm

Lemon peel, orange Hemoglobin catalyzed IC50 , 122.0 ppm
peel and peanut hull peroxidation of linoleic acid IC50 , 68.8 ppm IC50 , 111 ppm

IP: Induction period; ILLO: Inhibition of lecithin liposome oxidation as increase in ABS535 nm ; NPIT: Net prolongation of
induction time (min); IC50 : Inhibitory concentration for 50% inhibition in the reduction of oxidation.
∗ Reference: (Pokorny and Korezak, 1996).

prooxidant activity: A reduction in original antioxidant proper-
ties through the formation of compounds with prooxidant prop-
erties would appear to be of considerable interest with regard to
low temperature or short time heat treatments.
The influence of baking conditions and dough supplements on
the amounts of the antioxidant pronyl-L-lysine in bakery prod-
ucts was investigated (Lindenmeier and Hofmann, 2004).These
studies revealed high amounts of the antioxidant in bread crust,
and only low amounts in the crumb. The amounts of pronyl-
L-lysine were found to be strongly influenced by the intensity
of the thermal treatment. Substituting 5% of the flour with the
lysine rich protein casein or with 10% of glucose increased the
amounts of the antioxidant by more than 200%. To gain first
insights into the influence of the baking conditions on the an-
tioxidant potential of rye/wheat bread, five sour dough breads
(bread A-E) were freshly baked with variations in temperature
and time of thermal treatment (Table 17). The antioxidant po-
tential of the ethanolic fractions isolated from the bread crusts
exceeded the activities measured for the flour and the corre-
sponding crumb isolates. In addition, these data show a close
relationship between the antioxidant activity of the crust and the
baking time and also the increase in baking temperature was
found to accelerate antioxidant production.
Application of an in vitro antioxidant assay to solvent
fractions isolated from bread crust, bread crumb and flour,
respectively by Lindermeier et al. (2002) revealed the highest
antioxidative potential for the dark brown, ethanol solubles of
the crust, whereas the corresponding crumb and flour fractions
showed only minor activities. To compare the antioxidant
potential of bread crust fractions, the efficiencies of the
soluble fraction I-III, isolated from flour, crumb, and crust in
inhibiting the peroxidation of linolenic acid were measured and
calculated as Trolox equivalents (TE). The dark brown ethanol
solubles (fractions II) of the crust showed the highest activity
(Lindenmeier et al., 2002).
Preparation of Natural Antioxidants
There is a big difference between the preparation of syn-
thetic antioxidants and natural antioxidants for application in
food products and processing. Synthetic antioxidants are prod-
ucts of pure substances of constant composition and are applied
as such or in well-defined mixtures with other pure substances.
Natural antioxidants are available from raw materials of vari-
able compositions. Both the contents of active substances (usu-
ally a mixture of several compounds) and the content of various
other compounds, either inactive or possessing negligible ac-
tivities are present. Most widely used natural antioxidants are
not exactly purely natural, but nature identical. This means that
their structure is the same as that of natural substances, but they

Table 17 Baking conditions chosen for the manufacture of

Table 16 Changes in antioxidants during roasting and baking∗ the self-prepared bread A-E∗

Type of Type of Type of Baking regime

process precursors products Bread type Temp,◦ C (time, min)

Caramelisation Sugars, ascorbic acid Chelating macromolecules A 260(8)–220(62)

Maillard reaction Sugar, amino acids Chelating macromolecules B 260(16)–220(124)
Strecker degradation Dicarbonyls, amino acids Dihydroheterocycles C 260(24)–220(186)
Oxidation Phenolics Quinones D 280(8)–240(62)
Hydrolysis Glycosides, esters Aglycones, phenolic acids E 300(8)–260(62)
∗ Reference: (Niki, 1996). ∗ Reference:(Lindenmeier et al., 2002).
 ANTIOXIDANTS IN BAKERY PRODUCTS
have been prepared by synthesis. They are supplied in a rela-
tively pure state, like other synthetic antioxidants. Tocopherol,
ascorbic acid and citric acid belong to this group (Pokorny and
Korezak, 1996).
Active food ingredients may be directly applied. Some
nature-identical antioxidants, such as α-tocopherol or β-
carotene are available on the market in a pure form or in defined
solutions so that they can be added very easily in the amount
desired. Many other food components possessing antioxidant
activities are used in their natural form, such as spices. The
preliminary processing of such food components may be dry-
ing (in case of leaves or stems) and milling of dried material
(such as seeds). Better flavoring effect is found when rosemary
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and sage are used in a mixture with sodium glutamate, protein
hydrolysates, garlic, and onion, than as a single spice.
Antioxidants may be prepared by extraction of food ingredi-
ents. The content of active antioxidants in natural materials is
usually rather low so that large additions would be necessary to
obtain a significant improvement in stability against oxidation.
Extraction of antioxidants with fat and oils involves extraction
using edible oil or fat and is a very simple method. Natural mate-
rial containing antioxidants, such as herbs and spices, is mixed
with fat and/or oils, and the mixture is left at room tempera-
ture or at a moderately increased temperature for a defined time,
for example overnight, with or without stirring. The mixture is
then filtered and the fat or oil containing dissolved antioxidants
is used directly in food preparation. Rosemary, sage, paprika,
nutmeg, or cocoa shells have been powdered and extracted with
edible oil. The advantage of this procedure is its simplicity and
safety, as no organic solvents are used for the extraction there-
fore no residual solvents are present. Extraction of antioxidants
with organic solvents involves extraction with organic solvents
and is another possibility. Solvents used are hexane, acetone,
ethyl acetate, and methanol. The solvents of intermediary po-
larity seemed to be preferable to either nonpolar or highly polar
solvents. Ethanol would probably be better than methanol as
eventual solvent residues would be less toxic. Extraction of an-
tioxidants with supercritical fluid carbon dioxide is a modern
method is extraction with gases, usually carbon dioxide, under
supercritical conditions. Extraction with carbon dioxide is rela-
tively selective, generally better than that of organic solvents. A
big disadvantage of supercritical extraction is the high operation
pressure, which requires expensive equipment.
CONCLUSION
Fat is an essential food component and imparts palatability to
food. In addition it also improves appearance, taste, and texture
of food. Fat gives satiety after consumption. Apart from all these
advantages, fat has a disadvantage that it gets oxidized rapidly
upon exposure to air, moisture, and sunlight giving rise to an
unacceptable rancid flavor.
Due to change in lifestyle of people, ready to eat foods are pre-
ferred among which bakery products play a major role. Bakery
25
products invariably contain fats and oils, which oxidize slowly
during storage and various oxidation products cause rancidity
and deterioration of the sensory properties of the food products.
Autooxidation of fats and oils in processed foods may be pre-
vented by the use of oxidation inhibitors or antioxidants. Use
of antioxidants is an age-old concept. Invention of synthetic an-
tioxidants and their high efficiency to inhibit oxidation of fat at
low concentrations leads to their profuse use in bakery products.
The common synthetic antioxidants used are Butylated hydroxy
anisole (BHA), Butylated hydroxy toluene (BHT), Tertiary butyl
hydroxy quinone (TBHQ), and Gallates etc. They are available
at low cost and easy to incorporate into food products.
However, in recent years, demand from consumers for the
use of natural products than synthetic has increased. At the same
time long-term safety of these synthetic antioxidants is question-
able. Considering this criterion, many experiments were con-
ducted on shelf life of bakery products by using natural antioxi-
dants. Some of the natural antioxidants such as α-tocopherol, β-
carotene, ascorbic acid are almost equivalent to synthetic antiox-
idants when shelf life and consumer acceptability is concerned.
Recently, plant species have received much attention as major
sources of biologically active substances including antioxidants,
antimutagens and anticarcinogens. Plant extracts obtained from
some fruits and vegetables have been reported to be effective
antioxidants.
The spices most commonly used in bakery products are cin-
namon, mint, nutmeg, mace, cloves, poppy seeds, and sesame
seeds. Apart from these major sources, certain other specific
sources such as vanillin, milk peptides, tea extracts, and cocoa
extracts are evaluated for their antioxidant activity and some
of them are currently used as antioxidants in food products es-
pecially meat products. Hence, there is an immense scope for
the use of these natural antioxidants in bakery products, which
needs further research in this aspect.
FUTURE PROJECTIONS
The above review covers information on how both synthetic
and natural antioxidants are used in bakery products. It has been
concluded that use of natural antioxidants has many advantages
than synthetic antioxidants and can effectively replace synthetic
antioxidants. Some of the future areas of research needed in the
field of natural antioxidants are listed below.
1. Use of natural antioxidants in bakery products to increase
shelf life.
2. Use of natural antioxidants as nutraceuticals in bakery prod-
ucts to improve health benefits.
3. Molecular interaction between natural antioxidants and bak-
ery ingredients.
4. Incorporation of natural antioxidants in various food products
for prevention of diseases.
5. Utilization of under exploited antioxidants for food preser-
vation.
 26 B. NANDITHA AND P. PRABHASANKAR
6. Molecular mechanism of natural antioxidants during food
processing.
7. Studies on encapsulation of natural antioxidants for food pro-
cessing.
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