Department of Food Science
College of Home Economics
CENTRAL MINDANAO UNIVERSITY
University Town, Musuan 8710
Maramag, Bukidnon
BROWNING REACTIONS
Mylene A. Anwar
The browning reaction can be defined as
the sequence of events that begins with
the reaction of the amino group of
amino acids, peptides, or proteins with a
glycosidic hydroxyl group of sugars; the
sequence terminates with the formation
of brown nitrogenous polymers or
melanoidins (Ellis 1959).
POLYPHENOL OXIDASE
• Polyphenol oxidase (PPO, EC
1.10.3.1, systematic name 1,2
benzenediol:oxygen
oxidoreductase) is also labeled as
phenoloxidase, phenolase,
monophenol and diphenol oxidase,
and tyrosinase.
Enzymatic Browning
It is catalyzed by the enzyme
Phenoloxidase enzymes catalyze the
oxidation of phenolic constituents to
quinones, which finally polymerize to
colored melanoidins (Marshall et al.
2000).
Melanoidins have been described as low molecular weight (LMW) colored substances that are able to
cross-link proteins via ε-amino groups of lysine or arginine to produce high molecular weight (HMW) colored polyphenol oxidase (PPO) (1,2 benzenediol;
melanoidins. Also, it has been postulated that they are polymers consisting of repeating units of furans oxygen oxidoreductase, EC 1.10.3.1), which is
and/or pyrroles formed during the advanced stages of The Maillard reaction and linked by polycondensation also referred to as phenoloxidase, phenolase,
monophenol oxidase, diphenol oxidase, and
reactions (Martins and van Boekel 2003).
tyrosinase.
1 2
?
1. It catalyzes the hydroxylation of
phenols with oxygen, to the
o-position adjacent to an existing
hydroxyl group.
2. Oxidation of the diphenol to
o-benzoquinones, which are
further oxidized to melanins
(brown to dark products)
 CONTROL OF BROWNING

Non-enzymatic Browning

• Figure 3-6 Loss of Lysine Occurring as a Result of Heating of Several Foods. Source: From J. Adrian, The Maillard Reaction. IV.
The reaction velocity and pattern are influenced by the nature of the Study on the Behavior of Some Amino Acids During Roasting of Proteinaceous Foods, Ann. Nutr. Aliment. (French), Vol. 21, pp.
reacting amino acid or protein and the carbohydrate. 129-147, 1967.
 According to Hurst (1972), the following five steps are involved in
the process:

1. The production of an N-substituted glycosylamine from an

aldose or ketose reacting with a primary amino group of an amino
acid, peptide, or protein.
• The mechanism as shown is
2. Rearrangement of the glycosylamine by an Amadori thought to involve addition of
rearrangement type of reaction to yield an aldoseamine or the amine to the carbonyl group
ketoseamine. of the open-chain form of the
sugar, elimination of a molecule
3. A second rearrangement of the ketoseamine with a second
mole of aldose to form a diketoseamine, or the reaction of an
of water, and closure of the ring.
aldoseamine with a second mole of amino acid to yield a diamino
sugar. • The rate is high at low water
content; this explains the ease of
4. Degradation of the amino sugars with loss of one or more browning in dried and
molecules of water to give amino or nonamino compounds. concentrated foods.
5. Condensation of the compounds formed in Step 4 with each
other or with amino compounds to form brown pigments and
polymers.
Figure 3-9 Amadori Rearrangement. Source: From MJ. Kort, Reactions Figure 3-11 Heyns Rearrangement. Source: From MJ. Kort,
Figure 3-7 Reaction Pattern of the Formation of Melanoidins from Aldose Sugars and Amino Compounds. Source: From I.E. of Free Sugars with Aqueous Ammonia, Adv. Carbohydrate Chem. Reactions of Free Sugars with Aqueous Ammonia, Adv.
Hodge, Chemistry of Browning Reactions in Model Systems, Agr. Food Chem., Vol. 1, pp. 928-943, 1953. Biochem., Vol. 25, pp 311-349, 1970. Carbohydrate Chem. Biochem., Vol. 25, pp. 311-349, 1970.

• The Strecker degradation of a-amino acids is a reaction

that also significantly contributes to the formation of
flavor compounds. The dicarbonyl compounds formed
Schiff base in the previously described schemes react in the
ketosamine 1,2-enolamine following manner with a-amino acids:

new Schiff base

3-deoxyosulose
hydroxymethyl furfural
• The amino acid is converted into an aldehyde with one
enolform less carbon atom (Schonberg and Moubacher 1952).
Some of the compounds of browning flavor have been
described by Hodge et al. (1972). Corny, nutty, bready,
and crackery aroma compounds consist of planar
unsaturated heterocyclic compounds with one or two
nitrogen atoms in the ring.
unsaturated osulose

• Figure 3-12 1,2-Enolization Mechanism of the Browning Reaction. Source: From D.T. Hurst, Recent Developments in the Study of
Nonenzymic Browning and Its Inhibition by Sulphur Dioxide, BFMIRA Scientific and Technical Surveys, No. 75, Leatherhead,
England, 1972.
 Factors that influence the browning reaction are : Study of the Maillard Reaction in Foods

After formation of the aldosylamine, dehydration reactions
result in the production of 4- to 6-membered ring
compounds. When the reaction proceeds under conditions
of moderate heating, fluorescent nitrogenous compounds
are formed. These react rapidly with glycine to yield
melanoidins.
• Figure 3-13 Proposed Browning Reaction Mechanism According to Burton and McWeeney (1964).
Source: From H.S. Burton and DJ. McWeeney, Non-Enzymatic Browning: Routes to the Production of
MeIanoidins from Aldoses and Amino Compounds, Chem. Ind., Vol. 11, pp. 462-463, 1964.
1. Temperature (At higher temperatures, the carbon content of the pigment increases and more pigment is formed per mole of carbon dioxide • Frying or grilling of meat and fish may generate
released. Color intensity of the pigment increases with increasing temperature) low (ppb) levels of mutagenic/carcinogenic
2. pH (The browning reaction is slowed down by decreasing pH, and the browning reaction can be said to be selfinhibitory since the pH heterocyclic amines via Maillard reaction. The
decreases with the loss of the basic amino group. The effect of pH on the browning reaction is highly dependent on moisture content. The formation of these compounds depends on
reaction rate is significantly affected by the pH of the system; it generally increases with pH (Namiki et al. 1993, Ajandouz and Puigserver
1999).
cooking temperature and time, cooking
3. Moisture level (When a large amount of water is present, most of the browning is caused by caramelization, but at low water levels and at pH
technique and equipment, heat, mass
greater than 6, the Maillard reaction is predominant. Browning occurs at low temperatures and intermediate moisture content; the rate
transport, and/or chemical parameters.
increases with increasing water content. The rate is extremely low below the glass transition temperature, probably because of limited
diffusion (Roos and Himberg 1994; Roos et al. 1996a, b). • Although the mechanism of acrylamide
4. Oxygen formation in heated foods is not yet clear,
5. Sulfur dioxide (It is thought that sulfur dioxide reacts with the degradation products of the amino sugars, thus preventing these compounds several authors have put forth the hypothesis
from condensing into melanoidins. A serious drawback of the use of sulfur dioxide is that it reacts with thiamine and proteins, thereby that the reaction of asparagine (Fig. 4.5), a
reducing the nutritional value of foods. Sulfur dioxide destroys thiamine and is therefore not permitted for use in foods containing this major amino acid of potatoes and cereals
vitamin)
(Mottram et al. 2002, Weisshaar and Gutsche
6. The nature of the sugars (Reactivity is related to their conformational stability or to the amount of open chain structure present in solution.
Pentoses are more reactive than hexoses, and hexoses more than reducing disaccharides. Nonreducing disaccharides only react after
2002), or methionine (Stadler et al. 2002) with
hydrolysis has taken place. The order of reactivity of some of the aldohexoses is: mannose is more reactive than galactose, which is more reducing sugars (glucose, fructose) via Maillard
reactive than glucose). reaction could be the pathway.

Control of the Maillard Reaction in Foods Caramelization

• Indicators proposed to control the early stages of the Maillard reaction during food • Methods of preventing browning could consist of measures intended to • The formation of the caramel
processing: the 2- furoylmethyl amino acids as an indirect measure of Amadori pigment can be considered a
compound formation. slow reaction rates, such as control of moisture, temperature, or pH, or
nonenzymatic browning reaction
removal of an active intermediate. Generally, it is easier to use an in the absence of nitrogenous
• Determination of the level of Amadori compounds provides a very sensitive inhibitor. One of the most effective inhibitors of browning is sulfur
indicator for early detection (before detrimental changes occur) of quality changes compounds. When sugars are
caused by the Maillard reaction as well as a retrospective assessment of the heat dioxide. The action of sulfur dioxide is unique and no other suitable subjected to heat in the absence
treatment or storage conditions to which a product has been subjected (Olano and inhibitor has been found. It is known that sulfite can combine with the of water or are heated in
Martínez-Castro 1996, del Castillo et al. 1999). concentrated solution, a series of
• Evaluating for Amadori compounds can be carried out through furosine carbonyl group of an aldose to give an addition compound:
reactions occurs that finally leads
[ε-N-(2-furoylmethyl)- L-lysine] measurement. This amino acid is formed by acid
hydrolysis of the Amadori compound ε-N- (1-deoxy-D-fructosyl)-L-lysine. to caramel formation.
 • The initial stage is the formation of anhydro sugars (Shallenberger and • Caramelization of reducing • This could involve formation of compounds such as isosacchrosan.
Birch 1975). carbohydrates starts with the opening
of the hemiacetal ring followed by
enolization, which proceeds via acid-
• Glucose yields glucosan (1,2-anhydro-oc-D-glucose) and levoglucosan and base-catalyzed mechanisms,
(1,6-anhydro-p-D-glucose); these have widely differing specific rotation, leading to the formation of isomeric
+69° and -67°, respectively. carbohydrates (Fig. 4.8). The
interconversion of sugars through
their enediols increases with
• These compounds may dimerize to form a number of reversion increasing pH and is called the Lobry
de Bruyn-Alberda van Ekenstein • Caramelization of sucrose requires a temperature of about 200°C. At 16O°C,
disaccharides, including gentiobiose and sophorose, which are also sucrose melts and forms glucose and fructose anhydride (levulosan).
transformation (Kroh 1994).
formed when glucose is melted.

At 200°C, the reaction sequence consists of three distinct stages well
separated in time.
1. The first step requires 35 minutes of heating and involves a weight loss
of 4.5 percent, corresponding to a loss of one molecule of water per
molecule of sucrose.
2. After an additional 55 minutes of heating, the weight loss amounts to
9 percent and the pigment formed is named caramelan. This
corresponds approximately to the following equation:
3. A further 55 minutes of heating leads to the formation of caramelen. This ASCORBIC ACID BROWNING
compound corresponds to a weight loss of about 14 percent, which is about eight
molecules of water from three molecules of sucrose, as follows:
• Browning of ascorbic acid can be briefly
defined as the thermal decomposition of
4. Additional heating results in the formation of a very dark, nearly insoluble pigment ascorbic acid under both aerobic and
of average molecular composition C125H188O80. This material is called humin or anaerobic conditions, by oxidative or
caramelin.
nonoxidative mechanisms, in either the
The typical caramel flavor is the result of a number of sugar fragmentation and presence or absence of amino compounds
dehydration products, including diacetyl, acetic acid, formic acid, and two (Wedzicha 1984).
degradation products reported to have typical caramel flavor by lurch and Tatum
(1970), namely, acetylformoin (4-hydroxy-2,3,5-hexane-trione) and 4-
hydroxy-2,5-dimethyl-3(2H)-furanone.
 Pathway of Ascorbic Acid
• Factors that can influence the nature of the degradation mechanism include:
Browning
1. Temperature
2. salt and sugar concentration • When oxygen is present in the system, ascorbic acid is
degraded primarily to DHAA. Dehydroascorbic acid is not
3. pH stable and spontaneously converts to 2,3-diketo-L-gulonic
4. Oxygen
acid (Lee and Nagy 1996). Under anaerobic conditions,
DHAA is not formed and undergoes the generation of
5. Enzymes diketogulonic acid via its keto tautomer, followed by β
elimination at C-4 from this compound and
6. metal catalysts decarboxylation to give rise to 3-deoxypentosone, which
is further degraded to furfural. Under aerobic conditions,
7. amino acids xylosone is produced by simple decarboxylation of
8. oxidants or reductants diketogulonic acid and is later converted to reductones. In
the presence of amino acids, ascorbic acid, DHAA, and
9. initial concentration of ascorbic acid, and their oxidation products furfural, reductones, and
10. the ratio of ascorbic acid to dehydroascorbic acid (DHAA; Fennema 1976). 3-deoxypentosone may contribute to the browning of
foods by means of a Maillard type reaction (Fennema
1976, Belitz and Grosch 1997).
• A good prediction of ascorbic acid degradation and the evolution of the
browning index of orange juice stored under anaerobic conditions at 20–45°C
may be performed employing the Weibull model (Manso et al. 2001).
• Enzymatic oxidation also has been proposed as a mechanism for the
destruction of ascorbic acid in orange peels during preparation of
• Furfural, which is formed during anaerobic degradation of ascorbic acid, has a
marmalade. Boiling the grated peel in water substantially reduces the significant relationship to browning (Lee and Nagy 1988); its formation has
loss of ascorbic acid (Fennema 1976). Tyrosinase (PPO) may also been suggested as an adequate index for predicting storage temperature
abuse in orange juice concentrates and as a quality deterioration indicator in
possess ascorbic oxidase activity. single-strength juice (Lee and Nagy 1996).
• However, furfural is a very reactive aldehyde that forms and decomposes
simultaneously.
• The presence of metals, especially Cu2+ and Fe3+, causes great losses of
Vitamin C. Catalyzed oxidation goes faster than spontaneous oxidation.
• Anaerobic degradation, which occurs more slowly than uncatalyzed oxidation,
is maximum at pH 4 and minimum at pH 2 (Belitz and Grosch 1997). Ascorbic
acid oxidation is nonenzymatic in nature, but oxidation of ascorbic acid is
sometimes catalyzed by enzymes. Ascorbic acid oxidase is a copper-containing
enzyme that catalyzes oxidation of vitamin C. The reaction is catalyzed by
copper ions. The enzymatic oxidation of ascorbic acid is important in the citrus
industry. The reaction takes place mainly during extraction of juices.
Therefore, it becomes important to inhibit ascorbic oxidase by holding juices
for only short times and at low temperatures during the blending stage, by
deaerating the juice to remove oxygen, and finally by pasteurizing the juice to
inactivate the oxidizing enzymes.
Control of Ascorbic Acid Browning
• Sulfites (Wedzicha and Mcweeny 1974, Wedzicha and Imeson 1977), thiol compounds (Naim et al.
1997), maltilol (Koseki et al. 2001), sugars, and sorbitol (Rojas and Gerschenson 1997b) may be
effective in suppressing ascorbic acid browning.
• In general, the largest losses for noncitrus foods will occur during heating (Fennema 1976).
• The rapid removal of oxygen from packages is an important factor in sustaining a higher concentration
of ascorbic acid and lower browning of citrus juices over long-term storage.
• The extent of browning may be reduced by packing in oxygen-scavenging film (Zerdin et al. 2003).
Modified-atmosphere packages (Howard and Hernandez-Brenes 1998), microwave heating (Villamiel
et al. 1998, Howard et al. 1999), ultrasound assisted thermal processing (Zenker et al. 2003), pulsed
electric field processing (Min et al. 2003), and carbon dioxide assisted high-pressure processing (Boff
et al. 2003) are some examples of technological processes that allow ascorbic acid retention and,
consequently, a prevention of undesirable browning.