MAILLARD REACTIONS 1657
M
Macroelements in Milk and Dairy Products, Nutritional Signi®cance see Minerals in
Dairy Products: Macroelements, Nutritional Signi®cance.
MAILLARD REACTIONS
H Nursten, University of Reading, Reading, UK
Copyright 2002, Elsevier Science Ltd. All Rights Reserved
Chemistry
There are three main mechanisms by which non-
enzymic browning occurs in foods: the Maillard
reaction, caramelization, and ascorbic acid ox-
idation. The ®rst is the most important, involving
a reactive aldehyde (usually a reducing sugar) and
an amine (usually an amino acid, peptide or protein).
The second is undergone by sugars without amines,
but, in consequence, normally requires a consider-
ably higher temperature. The third is based on as-
corbic acid, which acts as a particularly reactive
sugar, but can also be oxidized enzymically and so
leads on to polyphenolic compounds, the normal
type of substrate for enzymic browning in foods.
Enzymic browning is virtually of no consequence
for dairy products, but is important for beverages,
such as tea, coffee and cocoa, which are likely to
come into contact with milk and dairy products.
The chemistry of the Maillard reaction is very
complex and can be thought to comprise cara-
melization and ascorbic acid oxidation as special
cases. Even a simple example of the Maillard reac-
tion, that between glucose and ammonia, leads to
more than 15 compounds and glucose and gly-
cine give more than 24, and that is without the
use of modern chromatographic methods. Using
high-performance liquid chromatography (HPLC)
and thin-layer chromatography (TLC) on solvent-
soluble material only (which constitutes as little as
0.1% w/w of the reactants), about 100 components
can be detected as reaction products from xylose
and glycine.
In order to understand something so complex,
simpli®cation is required. This was achieved remark-
ably successfully in 1953 through Hodge's scheme
(see Figure 1).
Hodge subdivided the Maillard reaction as
follows:
I. Initial stage: products colourless, without ab-
sorption of UV radiation (about 280 nm).
A. Sugar±amine condensation
B. Amadori rearrangement
II. Intermediate stage: products colourless or yel-
low, with strong absorption of UV light.
C. Sugar dehydration
D. Sugar fragmentation
E. Amino acid degradation (Strecker degradation)
III. Final stage: products highly coloured.
F. Aldol condensation
G. Aldehyde±amine condensation and formation
of heterocyclic nitrogen compounds.
Others have called the three stages Early, Advanced
and Final Maillard Reactions, respectively. The way
these reactions ®t together is outlined in Figure 1.
The ®nal products of nonenzymic browning are
called melanoidins to distinguish them from the
melanins produced by enzymic browning. Theoret-
ically, the distinction is clear, but in practice it is
1658 MAILLARD REACTIONS

+amino compound –H2O

N-SUBSTITUTED
ALDOSE
A GLYCOSYLAMINE

B
Amadori
rearrangement

H
1-AMINO-1-DEOXY-2-KETOSE

C D
–3 H2O
C –2 H2O

SCHIFF'S BASE
OF HMF OR FISSION PRODUCTS
FURFURAL (acetol, diacetyl,
REDUCTONES pyruvaldehyde, etc.)

+2 H –2 H
F
SUGARS –amino
compound E
+H2O DEHYDROREDUCTONES
Strecker
degradation
E +amino
HMF OR acid
FURFURAL F –CO2
G
F ALDEHYDES
F

ALDOLS AND
G + amino
N-FREE POLYMERS
G G compound

+amino +amino +amino

compound
G compound compound
+amino
compound

MELANOIDINS
BROWN NITROGENOUS POLYMERS AND COPOLYMERS

Figure 1 Nonenzymic browning. HMF, hydroxymethylfurfural. (Based on Hodge, 1953.)

very dif®cult to classify the dark brown products A. Sugar±Amine Condensation

formed in foods, since they tend to be very complex
This reaction is reversible. The amine can be a pro-
mixtures and chemically relatively intractable.
tein and, for example, it has been shown that insulin
Oxygen plays an essential part in enzymic
will react with glucose at a signi®cant rate even at
browning but it is not essential for nonenzymic
room temperature.
browning. It may help in fact, for example, in the ox-
There is no fundamental reason why the glycosyl-
idation of reductones, such as ascorbic acid to
amine should not act as the amine for a further
dehydroascorbic acid, but it may also hinder the
molecule of aldose, thus giving a diglycosylamine.
progress of the reaction, for example, in oxidizing
Lysine complexed as e-glycosylamine appears to be
pyruvaldehyde to pyruvic acid.
nutritionally available.
Table 1 lists 12 symptoms of nonenzymic brown-
ing and shows how these develop in relation to the
B. Amadori Rearrangement
three stages of it. Note, in particular that, as far as
browning itself is concerned, and also (off-)¯avour This reaction, which is thought to be acid catalysed,
production, there is an induction period. can be depicted as in Figure 2.
Hodge's scheme as outlined above, was subdivided It is important to note that, overall, the Amadori
into eight reactions, A to G (for H, see under G), each rearrangement is not reversible. The reaction goes
of which will be considered further in turn. spontaneously even at 25  C. Support for the above
 MAILLARD REACTIONS 1659

Table 1 Nonenzymic browning

Symptoms Stage

Initial Intermediate Final

1. Browning ÿ ‡ ‡‡‡
2. (Off-) ¯avour production ÿ ‡ ‡‡
3. Production of water ‡ ‡ ‡
4. Production of carbon dioxide ? ‡ ?
5. pH lowering ? ? ?
6. Reducing power (antioxidant activity) ‡ ‡ ‡
7. Solubility loss ÿ ÿ ‡
8. Loss of vitamin C ‡ ÿ ÿ
9. Loss of biological value of protein ‡ ‡ ‡
10. Chelation of metals ÿ ? ?
11. Toxicity ÿ ÿ ?
12. Fluorescence ÿ ‡ ÿ

+
RNH RNH RNH RNH

CH CH CH CH2
+H+
–H+
(CHOH)n–1 O CHOH COH CO

CH (CHOH)n–1 (CHOH)n–1 (CHOH)n–1

CH2OH CH2OH CH2OH CH2OH

N -substituted cation of enol form keto form

glycosylamine Schiff's base N-substituted 1-amino-2-deoxy-2-ketose

Figure 2 Amadori rearrangement leading to the Amadori compound N-substituted 1-amino-2-deoxy-2-ketose.

mechanism comes from the fact that, if the hydroxy
group on C2 is blocked by, e.g. a methyl group,
rearrangement becomes impossible. Further support
comes from the fact that 11 fructoseamino acids
and two difructoseamino acids have been found in
stored, freeze-dried apricots and peaches and some
have also been detected in dehydrated carrots,
cabbage, spray-dried tomato powder, tea (glutamic
acid, theanine), beet molasses, liquorice, roasted
meat and pig's liver extracts.
Compared with the N-substituted glycosylamines,
the 1-amino-1-deoxy-2-ketoses are more stable to
moist acid atmospheres, but are still heat labile
and decompose rapidly in mild alkali. They exert
greater reducing power, though less than reduct-
ones. They brown more easily with amino acids.
Acid hydrolysis gives much, compared with little,
hydroxymethylfurfural (HMF), but no hexose is re-
covered, in keeping with the reaction's being
irreversible.
Ketoses undergo a similar series of reactions, leading
to 2-amino-2-deoxyaldoses (Heyns rearrangement).
However, browning reactions of fructose differ from
those of glucose, e.g. loss of amino acids or of free
amino groups (casein) is much lower. It is worth
noting that lactulose stimulates the growth of bi®do-
bacteria in the gut.
Even when milk is exposed to only very mild
heating (60  C, 20 s), b-lactoglobulin is modi®ed,
whereas both a-lactalbumin and b-casein are unaf-
fected. On HPLC, b-lactoglobulin then appears not
only as its two genetic variants, A and B, but each
has a higher molecular mass component following
it. When HPLC is combined with electrospray mass
spectrometry (MS), the difference in mass for each
pair was shown to be 324 Da, corresponding to the
lactulosyl group. The lactulose has been found to be
attached to Lys47. When the Maillard reaction was
made more intense by storing the freeze-dried reac-
tion mixture at 65% rh, 2±11 lactulose molecules
became attached in 22 h at 50  C, with 1±7 in the
®rst 10 h. Lactulose became attached to 14 of the
15 lysine residues (not Lys101) and to the a-amino
group, 6.5 lactulosyl groups being bound per
1660 MAILLARD REACTIONS

b-lactoglobulin molecule on average at 50  C, but
only 0.5 in the more familiar aqueous systems. The
order of reactivity was Lys47, 91 (t ˆ 0 h)>a-amino,
Lys15, 70, 100 (t ˆ 2 h)>Lys60, 69, 75, 77, 83, 135,
138 (t ˆ 6 h)>Lys8, 141 (t ˆ 10 h).
Beta-lactoglobulin can also clearly be seen in its
two genetic variants by capillary electrophoresis.
Beta-lactoglobulin from skim milk powder exhibits
not just such a double peak, but a broad tail,
which can be resolved into three or more progress-
ively weaker pairs of peaks. By electrospray MS
the difference in mass between successive pairs has
been shown to be 324 Da, corresponding, as above,
to the attachment of a lactulose moiety. Native
b-lactoglobulin has been shown by capillary iso-
electric focusing to fall from 70% to <20% of total
b-lactoglobulin of the whey protein fraction when
milk powder was kept for 1 day at 37  C and
86% rh, making this a fast and convenient method
for monitoring storage conditions.
Examination of milk and dairy products by such
methods has far-reaching implications for elucid-
ating and controlling quality parameters. One
of the most important is that lysine locked up in
e-Amadori compounds becomes nutritionally not
available; this poses considerable analytical prob-
lems (see below).
C. Sugar Dehydration
There are two ways in which this occurs: under
acid conditions, furfurals are produced, whereas in
neutral or alkaline conditions and/or in the presence
of amines in nearly anhydrous systems, 6-carbon and
other reductones are produced (see Figure 3).
Furfural formation Various compounds can accel-
erate furfural formation, for example, glycine accel-
erates both the conversion of xylose to furfural
||
and that of glucose to HMF. The reason seems
to be that the Amadori product dehydrates more
readily than the original aldose or N-substituted
glycosylamine, giving the Schiff's base of furfural,
which is then hydrolysed, reliberating part of the
amine, but also condensing to melanoidins. It is gen-
erally thought that HMF has low browning potential
and is not on the main pathway to melanoidins.
Reductone formation Reductones can be thought
of as products formed from sugars by the loss
of only two molecules of water as compared with
the loss of three that leads to furfurals. Reduct-
ones are compounds which contain the group ±CO±
C(OH):C(OH)±, as in ascorbic acid, and a hexose
can readily be converted on paper to the vinylogue of
a reductone. Compounds such as reductones explain
the reducing power which develops during brown-
ing, but they take part in browning in the dehydro
form and therefore need oxygen to be converted to
it. Like furfurals, they brown more readily in the
presence of amines.
D. Sugar Fragmentation
The mechanism by which sugar fragmentation oc-
curs is accepted to be principally retroaldolization,
though oxidative ®ssion is also thought to play
a role. It should be recalled that retroaldolization
is an important part of the Embden±Meyerhof
glycolytic pathway (EMP), where fructose-1, 6-
diphosphate is split into dihydroxyacetone-phos-
phate and glyceraldehyde-3-phosphate. The sort of
reactions that occur is illustrated in Figure 4.
Fragments which retain the a-hydroxymethyl-
carbonyl grouping will undergo browning alone
in aqueous solution, but this will be greatly
accelerated by the presence of amines. The relative
reactivities of a-hydroxymethylcarbonyl compounds

||

CH[N CH\N –amine CH\O CH\O

| | |
\

+ H2O
C[OH –OH
– C[OH C\O –H2O
C\O 5-hydroxymethyl-
| | |
\

1,2-enolization furfural
CHOH CH +amine CH2 CH
low pH
| | | –H2O
\

–H2O
CHOH CHOH CHOH CH
| | | | +amine
+amine
CH2[N –H2O
||

1,2-enaminol 3-deoxyhexosulose
|
C\O
| melanoidins
C[HOH
|
C[HOH
| CH2[N
||

CH2 CH3 CH3 –H2O

| | | +amine
\

+amine
2,3-enolization C[OH C[OH C\O C\O fission
–amine
| | |
\

high pH
furanones,
C[OH C\O C\O C[OH
| | | C -methyl reductones
\

CHOH CHOH CHOH C[OH and α-dicarbonyls

| | | |
2,3-enediol 1-methyl1-2,3-dicarbonyl a reductone

Figure 3 Maillard reactions: the two major pathways from Amadori compounds to melanoidins. (Based on Hodge, 1967.)
 MAILLARD REACTIONS 1661

NHR NHR
| | CH2OH
CH2 CH2 –RNH2 |
| | C\O dihydroxyacetone
CO CO +H2O |
| | CH2OH
CHOH retroaldolization CH2OH
|
CHOH +
| CHO
CHOH |
| CHOH glyceraldehyde
CH2OH |
Amadori compound
CH2OH CH3
| acetic acid
COOH
CH3 oxidative fission +
|
CO COOH glycollic acid
| |
2,3-enolization CO CH2OH
|
CH2OH COOH
+ saccharinic | acetic acid
CH3
CHO rearrangement
etc. |
CH2OH CH3
glycolaldehyde | acetic acid
COOH
retroaldolization +
CH3
| COOH
CO oxidative fission |
| CHOH
CO |
| CHOH
CHOH |
| CH2OH
CHOH OH
| dehydration
isomaltol
CH2OH
(furan formation)
1-deoxyosulose COCH3
dehydration
O
(pyran
formation) O CH3
maltol
rearrangement
OH
O

CH3 2 mol CH3.CO.CHO pyruvaldehyde

| hydrolysis
CO
| CH3.CO.CO.CH2OH + CH3.COOH acetic acid
CO
|
CHOH CH3.CO.CO.CHO + CH3.CHO acetaldehyde
|
CO
| CH3.CO.CH2OH + CH3.CO.COOH pyruvic acid
CH3
acetol
diacetylformoin

Figure 4 Some examples of sugar fragmentation.

and other sugar fragments are given in decreasing
order in Table 2.
E. Strecker Degradation
This is a reaction of a-amino acids, in which they
are oxidized to the corresponding aldehyde, carbon
dioxide is given off, and ammonia is transferred
to other components of the system, very little
being liberated as such. The reaction is initiated by
compounds such as a-dicarbonyl compounds and
their vinylogues, or compounds which can give rise
to them readily, such as reductones by dehydrogen-
ation or imino analogues by hydrolysis. The reaction
may therefore by represented as follows:
R:CHNH2 :COOH ‡ R0 :CO:CO:R0 !
R:CHO ‡ CO2 ‡ R0 :CHNH2 :CO:R0
Studies with radioactive carbon have shown that
the carbon dioxide liberated in the Maillard reac-
tion does indeed originate from amino acids.
1662 MAILLARD REACTIONS

Table 2 Sugar fragmentation products in decreasing order of reactivity

Sugar fragmentation product Formula Reactivity

Glycolaldehyde CH2OH.CHO highest

Glyceraldehyde CH2OH.CHOH.CHO Ð
Pyruvaldehyde CH3.CO.CHO Ð
Acetol CH3.CO.CH2OH Ð
Dihydroxyacetone CH2OH.CO.CH2OH Ð
Acetoin CH3.CHOH.CO.CH3 Ð
Diacetyl CH3.CO.CO.CH3 Ð
Acetaldehyde CH3.CHO slightly lower
Aldol CH3.CHOH.CH2.CHO still lower
Propionaldehyde CH3.CH2.CHO very low
Pyruvic acid CH3.CO.COOH even lower
Levulinic acid CH3.CO.CH2.CH2.COOH Ð
Saccharinic acid e.g. CH2OH.(CHOH)2.CH2.CHOH.COOH not at all
Lactic acid CH3.CHOH.COOH Ð
Acetic acid CH3.COOH Ð
Formic acid H.COOH Ð
Formaldehyde H.CHO inhibits

The Strecker degradation enters browning reac-
tions in two ways: on the one hand, the aldehyde
formed can take part in aldol condensation leading
to nitrogen-free polymers or can react with amino
compounds to give melanoidins via aldimines, but
this is thought to be not a major colour-producing
reaction, because glycine can at times give more
browning with sugars than alanine, yet glycine pro-
duces formaldehyde by the Strecker degradation,
which has a negative effect on browning. Further-
more, amino acids other than the a-ones can also
produce melanoidins, but cannot undergo the
Strecker degradation. On the other hand, the dehydro-
reductones, derived from Amadori products by
dehydration and dehydrogenation, or the dicarbonyl
®ssion products, pick up the amino acid nitrogen
and go on to form melanoidins.
Dehydroascorbic acid (but see later) and quinones
formed enzymically from polyphenols can also take
the part of the dicarbonyl compound in the reac-
tion. Diacetyl is a well known fermentation product.
Being volatile, the aldehydes formed in the
Strecker degradation have often been thought to be
important contributors to the aroma of foodstuffs
and many patents have been granted for `reaction
¯avours', which use the Strecker degradation to
produce ¯avouring materials of various types, such
as maple, chocolate, coffee, tea, honey, mushroom
and bread.
F. Aldol Condensation
Aldehydes can arise by Reactions C, D and E, and
they can then react with each other by the aldol
condensation. Amines (and particularly their salts),
including peptones and egg albumin, are effective
catalysts. Additional carbonyl compounds which
can participate in the condensation may be derived
by the oxidation of fats.
Browning has been demonstrated for pyruval-
dehyde alone, furfurals, and pyruvate plus furfural.
Less browning occurs with sugars or with aldol itself.
Two molecules of diacetyl can undergo a similar
reaction twice to form 2,5-dimethyl benzoquinone.
Benzoquinones can act as dicarbonyl components
in the Strecker reaction, they readily form imines
in the cold, and they can be involved in the pro-
duction of melanoidins.
G. Aldehyde±Amine Condensation
Aldehydes, particularly a,b-unsaturated ones, react
readily at low temperature with amines to give
`polymeric' high-molecular-mass, coloured products
of unknown structure. Heterocyclic rings, such as
those of pyridines, pyrazines, pyrroles and imida-
zoles, have been shown to be present. Melanoidins
usually contain 3±4% nitrogen.
The constitution of melanoidins differs somewhat,
depending on how they have been produced, for
example:
furfural ‡ glycine ! high ether content
glucose ‡ glycine ! high alcoholic content
pyruvaldehyde ‡ glycine ! high enolic hydroxyl
and low ether content
As the condensations proceed, so higher molecular
mass products are formed, long-period products being
nondialysable. The results in Table 3 bear on this.
1. In relation to high molecular mass material, tem-
perature seems much more important than time.
 MAILLARD REACTIONS 1663

Table 3 Composition of retentate of melanoidins from xylose and glycine (molar ratio 1 : 1)

Reaction conditions Yield (%)a Microanalytical data (%)


Temperature ( C) Time Retentate Diffusate C H N

22 9 months 4.2 Ð 50.3 5.3 8.0

68 6 weeks 39.6 30.0 57.0 6.1 7.3
100 38 hours 30.7 15.8 57.8 5.4 6.7
a
Yield based on total reactants. Membrane cut-off, 12 kDa.

The proportion of high-molecular-mass material It therefore seems that Amadori compounds can
increased with temperature. undergo 1,2-enolization much more easily and
2. Loss of material, presumably H2O and CO2, is under milder acid conditions than the original sugar.
much greater at 100  C (54%) than at 68  C Under weakly acid conditions, some 2,3-enolization
(30%). occurs.
3. The composition of the retentate formed at 22  C It is probable that the Amadori forms of the
corresponds quite closely to the loss of 3 mol 1,2-enediol, of its derivative formed by loss of a
H2O: C, 49.1; H, 5.3; N, 8.2%, i.e. all N appears hydroxyl ion, and of the 3-deoxyhexosulose (norm-
to be retained. N is lost subsequently, some ally unstable to strong acid) are all relatively
presumably as volatiles. more stable and/or have longer lifetimes (Figure 3),
4. It is not clear to what extent, if any, these high- thus allowing them to equilibrate and to undergo
molecular-mass materials are coloured. side reactions.
4-Hydroxy-5-methyl-3(2H)-furanone is an im-
Melanoidins are also being studied in other ways.
portant contributor to cooked beef ¯avour, though
The 13C nuclear magnetic resonance (NMR) spec-
it has a caramel-type odour. It can be obtained
trum of high molecular mass water-soluble mater-
by heating xylose, ribose or ribose-phosphate with
ial from glucose and glycine has been found to be
amine salts and is formed by 2,3-enolization and
very like that of the corresponding Amadori com-
dehydration in a way similar to that shown in
pound, there being no evidence of unsaturated or
Figure 4 for the formation of isomaltol from a hex-
aromatic carbon atoms. The material is very dif®cult
ose. The furanone can be taken as symptomatic of
to hydrolyse, suggesting that the glucose units are
2,3-enolization just as furfural and HMF are symp-
linked by C±C bonds.
tomatic of 1,2-enolization.
The products of the interaction of a- or b-alanine
Table 4 shows clearly that low pH favours
with arabinose give rise to electron spin resonance
the formation of 2-furaldehyde (1,2-enolization),
(ESR) spectra with 17 and 23 lines, respectively.
whereas higher pH favours the formation of
These signals have been attributed to the presence
the furanone (2,3-enolization). It also illustrates
of the N,N 0 -dialkylpyrazine cation radicals. The rad-
how basicity of the amine can affect the route;
icals are detected before the Amadori compound
the least basic (dibenzylamine) is less likely to be
and hence it was thought that a new pathway (H)
protonated and therefore more liable to undergo
for browning had been discovered.
2,3-enolization.
Some of the above reactions have been carried
Use of 1-14C has shown that the methyl group is
out in D2O or tritiated (T2O, radioactive) water, to
derived from C1 of the hexuronic acid or the
study the extent of H exchange.
pentose. If prepared in D2O, NMR shows hydrogen
strong acid exchange from both C2 and CH3. This accords
D-glucose
(3 M HCl)
HMF no incorporation of D with CH2:C(OH) . . . intermediates (on the 2,3-enol-
into C-H bonds ization route).
strong acid
D-xylose
(3 M HCl)
furfural no incorporation of T Caramelization
into C-H bonds
Sugars, polysaccharides, polyhydroxycarboxylic
M HCl acids, reductones, a-dicarbonyl compounds and
Amadori compounds no incorporation at C3, but
from glucose some at C1 (aldehydic C-H)
quinones will undergo browning in the absence of
amino compounds.
2 M HOAc
strong incorporation at both Such reactions, even in the absence of a catalyst,
C3 and C1 are important in the food industry, but they require
1664 MAILLARD REACTIONS
Table 4 Conditions affecting 1,2-enolization and 2,3-enolization
Amadori form
2-Furaldehyde
Glucuronic acid ‡ 1 M dibenzylamine 16%
(derivative of fructuronic acid)
Glucuronic acid ‡ 1 M benzylamine 23%
(derivative of fructuronic acid)
Xylose ‡ 1 M benzylamine 41%
(derivative of xylulose)
high temperatures, not often encountered. For
example, glucose decomposes only above 150  C.
Caramelization is accelerated by carboxylic acids
and their salts, phosphates and metallic ions, but,
even when so catalysed, the energy requirements
exceed those of sugar±amine reactions.
As with the Maillard reaction, odorous com-
pounds are formed, water and carbon dioxide are
liberated, the pH drops during the reaction, colour
formation is markedly increased by increasing the
pH, oxygen has only a slight enhancing effect on
colour production, and the reaction is inhibited by
sulphur dioxide.
The main reactions are 1,2-enolization (Lobry de
Bruyn±Alberda van Ekenstein rearrangement, cf.
Amadori rearrangement), dehydration to furfurals
and ®ssion.
The volatiles produced by degradation of sugars
can make an important contribution to ¯avour.
Glucose decomposition at 300  C has been shown
to give more than 130 volatile degradation prod-
ucts, including maltol, which is also present in
a variety of food products.
Ascorbic Acid Oxidation
Ascorbic acid alone will brown in aqueous solu-
tion above 98  C, producing furfural and carbon
dioxide. Even in the presence of glycine, the
carbon dioxide comes essentially from ascorbic
acid (cf. Strecker degradation). The browning with
ascorbic acid also increases with pH and above
pH 7 autooxidation and browning occur even at
25  C. Other reductones will react similarly.
Glucose and fructose decrease the rate of browning
and so do amino acids initially, though later they
increase it.
Ascorbic acid is converted into furfural via
a pentose by loss of carbon dioxide. The dehydro
form ring opens into 2,3-diketogulonic acid, which
degrades readily, also leading to browning.
2 M H2SO4 at 100  C pH 7 at 100  C
Furanone 2-Furaldehyde Furanone
traces 0 present
0 0 present
0 0 present
Effect on Product Quality
The symptoms listed in Table 1 are closely linked to
qualities important in dairy products. They will
therefore be considered in turn.
Colour
Although colour is the named characteristic in
nonenzymic browning, much remains to be dis-
covered about this aspect of the Maillard reaction.
There are two fundamental questions: from where
does the colour come, i.e. what chromophore(s) is
(are) formed? and where does the colour reside, in
the high- and/or the low molecular mass material
formed? Contemporary workers are trying hard
to answer both questions.
As regards chromophore(s), the answer is de®n-
itely in the plural and a range of relevant compounds
identi®ed mainly among the low molecular mass
fraction of model Maillard systems is depicted in
Figure 5. Several of the molecules are worthy of
comment.
It should be noted that Compounds 1 and 2 are
built up of two C5 moieties, each derived, apparently
without chain fragmentation, from xylose, but one
is formed by 1,2-enolization and the other by 2,3-
enolization of the Amadori compound (see Figure 3).
Since these reactions require low and moderate
pH, respectively, pH of the medium must be a key
factor and the nature of the amino component of
the Amadori compound will play an important
role. The amino acid clearly forms a component of
Compound 1, but, as regards Compound 2, its role is
catalytic.
Compound 3 is built up from three C5 moieties,
but, whereas the central furanone moiety is bifunc-
tional, furfural is monofunctional and thus acts as
a capping group. It can be seen to ful®l the same
function in other compounds. Compound 4 is also
built up from three C5 moieties, but could readily
react further at its active methyl group.
 MAILLARD REACTIONS 1665

HO O O
HO O

O
O O HO
CH3 O X N

O
1 X = NCH2COOH orange (xyl + gly) 3 deep orange (xyl + gly) 5 yellow (xyl + gly or lys)
2 =O yellow (xyl + gly)

O HO O HO NCH3
O O

O CH3 O
N O O
O O
O 6 red (xyl + furfural
+ ala or val) 4 yellow (xyl + gly)
O O OH
O
7 intense orange (xyl + ala + furfural)

O
R
+ HOOC
N N N +
O CHO N N
H2N
N O
N OH
R
O H O
N O 10 intense yellow (pro + furfural)
O
COOH
9 deep red (N -acetylarg + glyoxal + furfural)

8 red-orange (furfural + ala)

Figure 5 The structures of some low molecular mass coloured compounds formed in model Maillard reactions. xyl, xylose; gly,
glycine; ala, alanine; val, valine; pro, proline; lys, lysine.

Compound 5 possesses a chain of eight carbon
atoms, which must involve fragmentation of the
xylose skeleton and reassembly of fragments (cf.
Figure 4). Here, the amino acid supplies the nitrogen
only and so both glycine and lysine have given rise
to it through interaction with xylose. Similarly,
alanine or valine supplies only the nitrogen for
Compound 6, for which four C5 units are required.
Compound 7 is built up from a C2 fragment and
three C5 units, but without the amino acid's nitrogen.
On the other hand, Compound 8 does involve
alanine along with four C5 units.
Other amino acids can give rise to different types
of chromophore. Thus, arginine can lead to Com-
pound 9 and proline (an imino acid) to Compound
10. Note that here furfural provides an open chain
of ®ve carbon atoms.
The above illustrates quite a range of chromo-
phores, but all of these were part of low molecu-
lar mass compounds. What about high molecular
mass compounds?
When 5 : 1 mixtures of glucose and glycine or
alanine in phosphate buffer, pH 7, are heated for
4 h at 95  C and separated by ultracentrifugation
through a series of membranes with progressively
smaller pore size, the molecular mass of almost
80% of the product is <1000 Da and nearly 20%
at 1000±3000 Da, representing 97±99% and 3±1%
of the recovered colour, respectively, i.e. very little
high molecular mass material has been formed and
by far the greatest proportion of the colour
resides in low molecular mass material. When the
amino acid is replaced by an equivalent amount of
b-casein, the result is very different: the molecular
mass of 43% of the product lies >100 kDa, 18% at
50±100 kDa, and 24% at <1000 Da, representing
90%, 9%, and <0.1% of the recovered colour, re-
spectively, i.e. over 60% of the material is >50 kDa
and represents virtually all of the recovered colour.
The b-casein (molecular mass 24 kDa) is cross-
linked into dimeric and progressively larger
entities, presumably through carbohydrate-derived
1666 MAILLARD REACTIONS

crosslinks between appropriate amino acid side
chains, as illustrated by some of the structures in
Figure 5. When sodium caseinate is heated with
lactose or glucose in milk-salts solution at 110±
150  C starting at pH 6.65, most of the browning is
also due to pigments bound to protein.
Volatile Compounds
About 3500 volatile compounds have been reported
to be formed by the Maillard reaction. Some ration-
alization is thus required. Accordingly, the volatile
products of the Maillard reaction can be arranged
in three groups, as listed in Table 5.
From a ¯avour point of view, pyrazines are the
most important compounds of Group 3. They are
derived from the dicarbonyl compound which
picks up ammonia in the Strecker degradation, as
shown in the equation in Section E above. The
mechanism is shown in Figure 6. Pyrazine formation
is favoured by higher temperature and pH, but
intermediate aw (0.75). Pyrroles can be formed by
replacement of the ring oxygen with ammonia or
amines in parallel to the nonvolatile but coloured
products included in Figure 5.
The Maillard reaction can lead to some non-
volatile compounds with bitter attributes, but these
have not so far been found to have signi®cance in
milk and dairy products.
aw
The Maillard reaction produces water and so
may increase aw, but the effect of aw on the Maillard

Table 5 Volatile products of the Maillard reaction

Group 1 `Simple' sugar dehydration/fragmentation products

Furans (e.g. HMF)
Pyrones (e.g. maltol)
Cyclopentenes (e.g. methylcyclopentenolone)
Carbonyls (e.g. CH3COCOCH3)
Acids (e.g. CH3COOH)

Group 2 `Simple' amino acid degradation products

Aldehydes, cf. Strecker degradation
Glycine CH2O
Alanine CH3CHO
Valine (CH3)2.CH.CHO
Leucine (CH3)2.CH.CH2.CHO
Isoleucine CH3.CH2.CH(CH3).CHO
Phenylalanine C6H5.CH2CHO
Tyrosine
Aspartic acid
Glutamic acid
Lysine
Arginine
Histidine
Tryptophan
Serine CH2OH.CHO
Threonine CH3CHOH.CHO
Cysteine CH2SH.CHO
Methionine CH3S.CH2CH2CHO, methional
Proline cannot
Hydroxyproline cannot
Sulphur compounds
Cystine H2S
Methional CH3SH ‡ CH2:CH.CHO

Group 3 Volatiles produced by further interactions

Pyrroles
Pyridines
Imidazoles
Pyrazines
Oxazoles
Thiazoles
Compounds ex aldol condensations

R NH2 O R9
HC C –2H2O
+
C CH
R9 O H2N R R9
2 M ex Strecker reagent
Figure 6 Formation of pyrazine from products of the Strecker degradation.
reaction is of much greater interest. Since loss of
water is part of the Maillard reaction, the mass
action effect of water at high aw will tend to hinder
the reaction, as will the dilution of the reagents.
On the other hand, as systems become more con-
centrated, mobility of the reagents is reduced.
Hence Maillard browning tends to exhibit a max-
imum at intermediate aw (e.g. whey powder at about
0.44, dried milk at 0.68).
pH and Loss of CO2
During the Maillard reaction, pH is lowered, both
by the production of acids (see Figure 4) and the
conversion of amines to cyclic nitrogen compounds,
though these changes would be counteracted to some
extent by the loss of carbon dioxide.
Antioxidant Activity (Reducing Power)
Most sugars have some reducing power and this in-
creases progressively as they are converted to glyco-
sylamines, Amadori compounds and reductones.
Maillard reaction products themselves also possess
antioxidant activity. In addition, their ability to com-
plex metals and to scavenge free radicals have rele-
vance here.
In suppressing hexanal formation through linoleic
acid oxidation, Maillard reaction products from
all the three sugars examined, fructose, glucose and
xylose, were very effective when combined with
histidine, which is a known metal chelator. Only
xylose±arginine produced a comparable result. The
products from lysine were somewhat less effective.
Equimolar amounts of the reactants are probably
to be preferred.
Ultra®ltration of the products led to the greatest
activity for the xylose±glycine fraction of about
4500 Da, its effect being greater than that of butyl-
ated hydroxyanisole (BHA), but less than that of
butylated hydroxytoluene (BHT) on an equal-weight
basis. There was a strong synergistic effect between
it and BHA. With glucose±histidine, the fraction
>1000 Da had six times the activity of that of the
crude reaction mixture.
Much still needs to be discovered in this area,
which is also of physiological importance, and con-
tinues to be an active subject of research.
MAILLARD REACTIONS 1667
R N R9 R N R9
–2H
N R R9 N R
a dihydropyrazine a pyrazine
Solubility Loss
Melanoidin formation is invariably accompanied by
an increase in molecular size, which almost always
results in a decrease in solubility. However, the
stability of milk to lowering of pH and to calcium is
improved by preheating, which is attributed to the
modi®cation of lysine residues and the consequently
increase in negative charge on casein.
Loss of Vitamin C
Ascorbic acid is a reductone and is involved in
nonenzymic browning, as has been outlined above.
Milk is a signi®cant source of vitamin C and any loss
would detract from its nutritional value.
Loss of Biological Value
Lysine is an essential amino acid and so its loss
through the Maillard reaction seriously detracts from
the nutritional value of milk, which is a good source
of it. Such loss becomes even more important in the
case of infant formulae.
Determination of nutritionally blocked lysine As
described in Section B, the Amadori rearrangement of
e-N-glucosyllysine irreversibly gives e-N-deoxy-
fructosyllysine (I). Thus, lysine has become nutri-
tionally no longer available from that point onwards.
Amino acid analysis of proteins normally involves
6 M HCl. The effect on I of hydrolysis with such
strong acid is shown in Figure 7. In amino acid
analysis by ion exchange, pyridosine elutes ahead
of lysine, and furosine elutes well after arginine.
Because of the partial recovery of lysine, interpret-
ing the results of analysis is complicated. It is import-
ant to appreciate that lysine locked up in the Amadori
compound, although partially recoverable by amino
acid analysis, is no longer available nutritionally.
When lactose reacts with protein in dairy prod-
ucts, lactulosyllysine residues are formed. On hy-
drolysis, these give yields of 40% recovered lysine
and 32% furosine. The lysine residues nutritionally
blocked can then be calculated as follows:
% lysine nutritionally blocked ˆ
3:1 furosine
 100
total lysine ‡ 1:87 furosine
1668 MAILLARD REACTIONS

H2N
6 M HCl
ε-N-deoxyfructosyllysine CH.(CH2)4.NH2 50%
24 h
HOOC
lysine

+ ---NH.CH2.CO O 20%

furosine

H3C

+ ---N O 10%

OH
pyridosine

Figure 7 The products of hydrolysis of e-N-deoxyfructosyllysine under protein analysis condition.

The factors are derived thus: 100/32 ˆ 3.1 and
60/32 ˆ 1.87. Total lysine represents the total lysine
recovered in the analysis, i.e. unreacted lysine
plus lysine recovered from lactulosyllysine residues.
Table 6 gives results that have been obtained in
this way for lysine damage in good manufactur-
ing practice. For UHT milks, the ratio of pyridosine
to furosine was 0.36. UHT milk with furosine
>50 mg lÿ1 should be regarded as overprocessed.
Ne-Carboxymethyllysine (see below and Figure 8)
correlates well with furosine (r ˆ 0.96), from which
it is probably derived by oxidation. Hydroxy-
methylfurfural, which is more readily determined,
by gas chromatography, correlates with furosine
equally well (r ˆ 0.96) for pilot-plant UHT milk, but
less well (r ˆ 0.85) for commercial UHT milk.
Determination of furosine in acid hydrolysates by
reversed-phase HPLC in conjunction with an ex-
ternal standard of 2-acetylfuran is about 10 times
as sensitive and leads to values more than double
those given above.
Milk and dairy products can also be hydrolysed
by a succession of selected enzymes and the result-
ant hydrolysates examined by amino acid analyser,
when lactulosyllysine elutes shortly after phenyl-
alanine. The results for percentage lysine modi®ed
obtained in this way for nine samples of different
infant formulae were 3±5.5 times as high as those
obtained by the furosine method. This reinforces the
view that the results given above are considerable
underestimates.
Lactose not only participates in the Maillard
reaction, but can also undergo a variety of other re-
actions, of which isomerization to lactulose (Lobry
de Bruin±Alberda van Ekenstein rearrangement) is
the most important. Accordingly, lactulose concen-
tration is a useful index of exposure to heat, even
though the smallest amount determinable is quite
Table 6 Lysine damage in good manufacturing practice
Process Lysine
damage (%)
Raw or freeze-dried milk 0
Pasteurized (74  C, 40 s) 0±2
UHT sterilized (135±150  C, a few s) 0±3
Spray-dried powder 0±3
Sweetened condensed 0±3
In-container sterilized ¯uid 8±12
Roller-dried (without precondensation) 10±15
Evaporated 15±20
Roller-dried (conventional) 20±30
From Mauron (1981).
high (30 mg lÿ1). Levels (mg lÿ1) found in milk are
as follows: pasteurized, 0±82; UHT direct, 41±670;
UHT indirect, 120±1430; sterilized, 412±1840. The
different classes of milk are therefore not clearly
differentiated by lactulose content alone. There
is little, if any, increase in lactulose on nor-
mal storage and lactulose does not appear to be
formed during milk drying, but lactulose correlates
extremely well with furosine for fresh UHT milk
(r ˆ 0.99). There is thus scope for the ratio of furo-
sine to lactulose to provide additional informa-
tion on the thermal history of a sample, particularly
of sterilized milk.
The e-N-deoxyfructosyl groups, which on acid
hydrolysis give rise to furosine and pyridosine,
can undergo a number of reactions (cf. Figure 4),
leading, for example, to acetic acid. If this were to
remain attached to the e-amino group, carboxy-
methyllysine (CML) would result (Figure 8). CML
levels can be determined by reversed-phase HPLC
with ¯uorescence detection and precolumn de-
rivatization with o-phthalaldehyde, reproducibil-
ity being very good (RSD 2.2%). CML was not
 MAILLARD REACTIONS 1669

HOOC.CH2.NHR HOCH2 CHO NHR9

N +
N NH
R
R
carboxymethyllysine pyrraline pentosidine

R = (CH2)4.CH(NH2)COOH
R9 = (CH2)3.CH(NH2)COOH

Figure 8 Some amino acids combined with intermediate Maillard products and determined in milk and dairy products.

detected in UHT milk, most ¯avoured milks, and all Chelation of Metals
cheeses (n ˆ 49), except for a whey cheese, which
Melanoidins have metal-chelating properties. Includ-
had the highest amount (1691 mg kgÿ1), followed
ing 10% of the Maillard reaction product (MRP)
by an evaporated milk (1015 mg kgÿ1, n ˆ 9, me-
from glucose±glutamate in the diet of rats has been
dian 499). A single sterilized milk sample out of 13
shown to lead to severe diarrhoea and nephro-
had 343 mg kgÿ1 protein. CML is questionable as a
calcinosis. Rats similarly fed 0.5% MRP did not
marker of heat damage in conventional milk prod-
show signi®cant changes in the retention of calcium,
ucts, whereas furosine allows evaluation of the early
magnesium, copper or iron, but zinc retention was
stages of the Maillard reaction. Determination of
reduced by about a third. Zinc is one of the metals
CML becomes useful for a range of products: se-
in which human diets tend to be marginal.
verely heat-treated samples, products where a long
shelf-life is required and where stability is achieved Toxicity
with salt additions, milk products made with com-
The content of 4-methylimidazole in ammonia
ponents that have already been compromised (e.g.
caramel has a legal limit of 200 mg kgÿ1. Abnor-
cocoa products), and other products, such as `follow-
malities associated with its ingestion by rats in-
on' milks.
clude lymphocytopenia, but its control is more as
HMF is an important breakdown product of
an indicator of good manufacturing practice than
hexoses via the Amadori compound and 1,2-enol-
because of a threat of toxicity. Mutagenicity in
ization. One can imagine its reacting with e-amino
Maillard products has been traced to compounds
groups to give pyrraline (Figure 8). Pyrraline can
such as imidazoquinolines and imidazoquinoxalines,
be determined by reversed-phase HPLC with elec-
the formation of which involves creatine and/or
trochemical detection and it has been shown to
creatinine, compounds present primarily in muscle
increase progressively in the deproteinized whey
foods.
fraction of non-fat dried milk over 6 h at 80  C
from 2 to 133 mg kgÿ1. It has also been found in a Fluorescence
range of processed foods, including a cheese gravy.
During storage of freeze-dried milk at 70  C, pyr- Although ¯uorescence has been noted in Maillard
raline (determined after enzymic hydrolysis) in- products, particularly at the intermediate stage, no
creased almost linearly with time and the amount systematic study has been made of it. Fluorescence
formed increased with moisture content, reaching has proved useful, especially in examining sec-
more than 5000 mg kgÿ1 protein in 50 h at 9% tions under the microscope. Several molecules de-
moisture. Values up to 3100 mg kgÿ1 protein were rived from crosslinks obtained under physiological
found in some samples of milk or whey powder, conditions ¯uoresce (Exmax 320±380, Emmax 380±
attributable to overprocessing and/or storage under 465 nm), providing speci®c data to supplement the
adverse conditions. relatively characterless visible absorption spectra.
Once the Maillard reaction has led to the conver- However, quenching of ¯uorescence provides a po-
sion of an e-sugar residue to an a-diketo com- tential drawback.
pound, such as a 3-deoxy-1,2-osulose, reaction with
an arginine residue becomes possible, crosslinking Practical Applications
the protein. Pentosidine (Figure 8) is a compound
Colour
formed in such a way. It has been found in small
amounts (2±5 mg kgÿ1 protein) in acid hydrolysates The development of colour due to the Maillard re-
of sterilized milk and in evaporated milk. actions is usually detrimental for dairy products and
1670 MAILLARD REACTIONS
therefore normally becomes a question of avoidance
or moderation. This is done by using where feasible
lower temperatures, shorter processing and storage
times, and lower pH, and avoiding intermediate aw.
Volatile Compounds
Individual samples of milk and dairy products have
been found to yield up to 400 volatile components,
most of which are derived from milk lipids or
microbial action. The Maillard reaction becomes
important when heat is applied or on longer storage.
Hydroxymethylfurfural is a product of the inter-
mediate stage of the Maillard reaction and it is
volatile. However, the total amount formed is
variable and does not correlate well with process-
ing conditions, values ranging from traces to
24.1 mmol lÿ1 over raw, UHT (direct and indirect),
and in-can sterilized milks. Nevertheless, HMF has
been used to follow successfully indirectly heated
UHT cow/buffalo (1 : 1) milk through storage at
37  C and 22  C, total HMF falling initially
from 18.3 to 15.0 and 12.8 after 9 days, in parallel
with the decrease in sulphydryl groups, and then
increasing progressively to 26.8 and 21.9 mmol lÿ1,
respectively, after 33 days, as residual oxygen con-
tinues to be depleted. The product became unac-
ceptable sensorily after about 25 days.
At least 67 volatile compounds can be detected in
the headspace above whey protein concentrate equili-
brated to 75% rh and kept under accelerated sto-
rage conditions of 70  C for 4 days. Most of the
33 compounds identi®ed were clearly lipid break-
down products, but 2-methylbutanal can be attrib-
uted to the Strecker degradation, 2-furanmethanol
to carbohydrate degradation, methyl- and 2,6-di-
methylpyrazine to the Maillard reaction (Group 3),
and dimethyl disulphide and trisulphide to meth-
ionine breakdown.
Block milk or white crumb is an intermediate
product in the manufacture of milk chocolate and
white chocolate. It is made from concentrated milk
and sugar and more than 30 volatiles have been
identi®ed in the headspace. Of these, again, most
are clearly lipid breakdown products, as with the
whey protein concentrate (above), but they also
included 2- and 3-methylbutanal, dihydro-2-methyl-
3(2H)-furanone, 2/3-furfural, 1-(2-furanyl)ethanone,
2/3-furanmethanol, pyrazine, methylpyrazine, pyr-
role, dimethyl disulphide and trisulphide, formic
acid and 2- and 3-methylbutanoic acid. Quantit-
ative descriptive analysis resulted in a list of nine
¯avour and ®ve taste attributes, including milk,
nutty, caramel and burnt. Considerable further
work is required.
Functionality
Furosine level is a useful indicator of heat treatment
and of storage conditions. With milk, there is little
change on pasteurization and 8.6 mg furosine
100 gÿ1 protein is thought to be a useful upper limit
for pasteurized, peroxidase-positive milk. Seven
commercial samples of such milk from Germany
gave values in the range 6.0±7.3 mg 100 gÿ1 protein.
Twenty-six samples of Italian pasteurized milk out of
124 examined gave higher results (some even ex-
ceeding 100 mg 100 gÿ1 protein), implying the pres-
ence of reconstituted milk powder.
UHT treatment brings about a more than ten-
fold increase over raw milk and in-bottle steriliza-
tion one that is more than 50-fold. Formation of
furosine continues on storage, for example, in pas-
teurized milk at 6±8  C at 1 mg 100 gÿ1 protein every
16 days.
With skim milk powder, there is again little sen-
sitivity to pasteurization conditions, but a greater
than tenfold increase on drying, during which the
whey protein nitrogen index (WPNI) changes little.
During drying, the powder passes through the region
of aw of maximum Maillard reactivity, resulting
in a range of furosine values from 55 mg 100 gÿ1
protein for extra low-heat to 350 mg 100 gÿ1 protein
for high-heat samples. Similar results have been ob-
tained for skim milk powder by determining lactulo-
syllysine after enzyme hydrolysis. Because of the
increased furosine content of milk powder, furosine
determination can indicate the adulteration of fresh
milk with reconstituted. The ratio of furosine to
lactulose can similarly indicate the adulteration of
UHT milk with reconstituted.
Furosine has been used to indicate the degree to
which lysine has been damaged in milk-based
foods with the following results: condensed milk,
36.3%; milk nougat, 34.3%; soft nougat, 33.0%;
milk chocolate, 27.1%; white chocolate, 18.5%;
chocolate cream, 14.8%; milk tablet 1, 13.4%; rice
cream, 11.2%; milk tablet 2, 11.0%; cooked
cream, 2.6%; yoghurt mousse, 2.6%; and dietetic
meal, 2.5%.
For most infant formulae, the proportion of lysine
blocked (13±27%) is not high enough to reduce
the lysine intake to below the FAO suggested
value of 102 mg kgÿ1 dayÿ1. The amount of lysine
blocked can be lowered by substituting lactose with
glucose syrup of dextrose equivalent <15. The
production of dietetic milks by enzymic hydrolysis
of lactose can double the concentration of redu-
cing sugars and is thus liable to increase the amount
of lysine blocked, but in none of the milks ana-
lysed was the level of free lysine residues lowered
 MAILLARD REACTIONS 1671

so far as to make lysine the limiting amino acid. It Fluorescence

is important that `lactose-free' milk, in particular, is
Two readings with ex and em at 290 and 340 and
stored <4  C.
350 and 440 nm, respectively, taken on the trans-
When skim milk powder was stored at 20±30  C
parent soluble fraction of milk at pH 4.6, can be
for 6 months, a further 15% lysine ceased to be
used to determine tryptophan and intermediate
available, as indicated by the dye-binding method.
Maillard reaction products, respectively. The former
During storage, whey protein solubility changes
is a marker of protein denaturation and thus of
little, whereas furosine level can more than double
low-temperature treatment of milk, such as in pas-
in 10 months, in accord with changes in rennet
teurization, whereas the latter is an indicator of
coagulation time and curd tension. Values of up to
high-temperature treatment, such as indirect UHT
1200 mg furosine 100 gÿ1 protein have been ob-
processing. From an examination of 80 samples,
tained for atomized skim milk powder on storage at
ranging from raw to indirect UHT-treated milk, it
30  C and 0.44 aw for 90 days.
was concluded that the method allows rapid dis-
Industrial production of Mozzarella cheese
tinction between the various types of milk and could
involves only limited exposure to heat and so does
form the basis of a rapid quality control system in
not produce furosine values greater than 8 mg
plants heat-treating milk.
100 gÿ1 protein. Data for European commercial
The relationship between ¯uorescence (ex
cheeses (n ˆ 73) gave 4±38 mg 100 gÿ1 protein, sug-
347 nm, em 415 nm) and heat treatment of milk
gesting some use of raw materials other than
has already been studied, using percentage relative
fresh liquid milk.
¯uorescence in the form of a ¯uorescence index.
Rennet casein is the protein of choice in the
Mean results for Spanish milks were as follows:
manufacture of Mozzarella cheese analogues
pasteurized, 20  2 (n ˆ 9); UHT, 28  4 (n ˆ 36);
having melt and ¯ow characteristics suitable for
in-bottle sterilized, 44  5 % (n ˆ 6). For the UHT
pizza toppings. Their functionality in this regard can
samples, there was a good linear correlation with
be very variable. The furosine level was found to
HMF content (r ˆ 0.935). The behaviour of the
range from 6.2 to 123.8 mg 100 gÿ1 protein and
¯uorescence on storage of the milks has not been
correlated with hydration characteristics, such as
studied and may well confound the results. In
time taken to reach maximum viscosity index. The
model systems, lactose in the absence of casein and
variability may be due to the temperature and dura-
whey proteins gave 500 to 1000 times less ¯uores-
tion of washing the curd and of drying, the heat-
cence than when either protein was present.
ing effects of grinding or milling, and the storage
From the above, it can be seen that the network
conditions.
of the Maillard reactions is well established, but
The MRPs of alanine/xylose (500 mg kgÿ1) and
that a great deal of detail needs to be ®lled in as
lysine/lactose (1000 mg kgÿ1) possess greater anti-
regards most of its aspects, chemical, quality-related
oxidant activity in relation to butter stored at 6  C
and applied.
than BHT (200 mg kgÿ1), whereas other MRPs,
such as that of alanine/xylose (1000 mg kgÿ1),
See also: Lactose: Properties, Production, Applications.
exhibited a slight prooxidant effect. Development of Lipids: Lipid Oxidation. Liquid Milk Products:
acidity was retarded most by the MRP of proline/ Sterilized Milk. Milk Powders: Physical and Functional
xylose and lysine/xylose (both 1000 mg kgÿ1). Properties. Milk Proteins: Beta-Lactoglobulin.
Nutritional Role of Dairy Products: Effects of
Toxicity Processing on Protein Quality of Milk and Milk Products.
The presence of imidazole mutagens has not been
reported for dairy products, though some muta- Further Reading
genicity will develop in dairy products as in many Hodge JE (1953) Chemistry of browning reactions in
other foods on severe heat treatment. There is no model systems. Journal of Agricultural and Food
mutagenic activity in HTST pasteurized milk, in Chemistry 1: 928±943.
UHT milk, or in in-bottle sterilized milk. In fact, Hodge JE (1967) Origin of ¯avor in foods: nonenzymatic
mutagen binding to casein may reduce the muta- browning reactions. In: Schultz HW, Day EA and
genicity of some food systems. Libbey LM (eds.) The Chemistry and Physiology of
Allergenicity of b-lactoglobulin may be enhanced Flavors, pp. 465±491. Westport: AVI.
by the Maillard reaction, so in manufacturing IDF (1991) Standard 147: Heat Treated Milk. Determina-
hypoallergenic milk formulae it is advisable to re- tion of Lactulose Content. High Performance Liquid
Chromatography (Reference Method). Brussels: Inter-
duce the concentration of lactose.
national Dairy Federation.
1672 MAMMALS

McSweeney PLH, Nursten HE and Urbach G (1997)
Flavours and off-¯avours in milk and dairy products.
In: Fox PF (ed.) Advanced Dairy Chemistry, vol. 3,
Lactose, Water, Salts and Vitamins, pp. 403±468.
London: Chapman & Hall.
Mauron J (1981) The Maillard reaction in food: a critical
review from the nutritional standpoint. In: Eriksson C
(ed.) Maillard Reactions in Food: Chemical, Physiolo-
gical and Technological Aspects, pp. 5±35. Oxford:
Pergamon Press.
O'Brien J (1997) Reaction chemistry of lactose: non-
enzymatic degradation pathways and their signi®cance
in dairy products. In: Fox PF (ed.) Advanced Dairy
Chemistry, vol. 3, Lactose, Water, Salts and Vitamins,
pp. 155±231. London: Chapman & Hall.
O'Brien J, Nursten HE, Crabbe MJC and Ames JM (eds.)
(1998) The Maillard Reaction in Foods and Medicine.
Cambridge: Royal Society of Chemistry.

MAMMALS

I A Forsyth, The Babraham Institute, Cambridge, UK
Copyright 2002, Elsevier Science Ltd. All Rights Reserved
Introduction
Mammals are de®ned by the ability to produce milk
for the nurture of their offspring. Living members
of the class Mammalia belong to one of three
groups: the egg-laying monotremes (the duck-billed
platypus and the echidna) in the subclass Proto-
theria and in the subclass Theriiformes (therians), the
marsupials and the eutherian (placental) mammals.
The basic structure of the mammary glands is
remarkably similar in all these groups, although
monotreme glands do not have nipples. Instead of
being suckled, the hatched young lick milk from
a specialized skin area.
The early mammal-like reptiles appear in the fossil
record (Table 1) during the Permian and Triassic
Periods. Mammals are thought to have evolved at
the close of the Triassic from small therapsid reptiles,
called cynodonts. These are found from the late
Permian and already share some bony features with
mammals, such as specialization of teeth and a hard
palate (see `Mammalian Characteristics', below).
Progression to producing milk is suggested to involve
the development of cutaneous glands in egg in-
cubation patches producing antimicrobial secretions
to ®rst protect the eggs and later supply nutrients
to the hatchlings. Part of the argument for this

Table 1 The geological time-scale, in millions of years ago (mya)

Era Period Epoch

Paleozoic Cambrian (540-505 mya) to Ð

540-245 mya Permian (286-245 mya)

Mesozoic 245-66 mya Triassic (245-210 mya) Ð

`Age of Reptiles' Jurassic (210-144 mya)
Cretaceous (144-66 mya)

Cenozoic 66 mya ± present Tertiary (66-1.8 mya) Paleocene (66-57 mya)

`Age of Mammals' Eocene (57-36 mya)
Oligocene (36-23 mya)
Miocene (23-5 mya)
Pliocene (5-2 mya)

Quaternary (1.8 mya ± present) Pleistocene (2 mya-10 000 ya)

Holocene (10 000 ya ± present)