Maillard browning

• The Maillard reaction is also known as non-enzymic

browning
The Maillard reaction: • Very different from enzymic browning which occurs when
Implications for the dairy industry an apple is cut
• Involves several reactions; the formation of brown
pigments is the last one.
Hilton Deeth • Maillard browning is seen in many cooked foods, e.g.,
(h.deeth@uq.edu.au) meat, bread, potato chips, roasted coffee

• Less visible in most milk and dairy products but still very
22 February 2017 important

The Maillard Reaction

The Maillard Reaction 2
• Named after Louis Camille Maillard who
published a paper on the brown pigments,
• Reaction requires heat; the higher the
melanoidins, in 1912
temperature, the faster it goes
• Starts with a reaction between a reducing sugar
• In the dairy industry it occurs during high-
and a protein
temperature processing
• [A “reducing” sugar is one that can be oxidised, in • A major reason why UHT processing replaced in-container
chemical terms one with an aldehyde group which sterilisation – sterilised milk has a brownish colour
can be oxidised to a carboxylic acid, e.g., lactose, • And, importantly, it also occurs during storage,
glucose, galactose but NOT sucrose] particularly at temperatures above ~30°C.
• Sucrose-sweetened products, e.g., sweetened condensed milk,
are not more susceptible to Maillard browning unless the sucrose • It occurs in powders as well as liquid products
contains some invert sugar (glucose+ fructose)
 Significance in food processing Factors affecting the Maillard reaction

“Among the various processing-induced chemical • Temperature

reactions in proteins, the Maillard reaction (non-
enzymic browning) has the greatest impact on • Amount and type of reducing sugar
sensory and nutritional properties” [Fennema, O.R. (1996) • Time
Food Chemistry (3rd edn)]
• Relative humidity (for powders)
Brown colour formation and flavour change are the most • pH – faster reaction at higher pH
significant effects
[WWII soldiers complained that dried egg was going brown
• Stabilisers used in sterilisation
and developing off-flavour]

Effects on milk and dairy products

Adverse effects
• Browning is the obvious effect but is only noticed in:
o very highly heated products, e.g., sterilised milk
o lactose-hydrolysed milk
 The products of lactose hydrolysis, glucose and galactose, react much
faster than lactose in the Maillard reaction
o products stored at high ambient temperature (>30C) for several
weeks or months, e.g., milk powders, UHT milk
Skim milk, goat’s milk, full cream cow’s milk (X2) and
lactose-reduced milk, after storage at 50 °C for 4 months
Deeth & Lewis (2017)
 Effects (cont)
• Off-flavour production, especially during storage of long-shelf-
life products
• Reduces pH in UHT milk during storage due to production of
formic and acetic acids
• Causes protein cross-linking which may reduce solubility of
some powders
• Reduction in nutrient value through blocked lysine
o a concern for products such as infant formulae
Beneficial effects
• Some Maillard products have antioxidant properties
• CSIRO scientists showed heated casein-glucose mixture had
Lactose-reduced milk samples after storage at 4, 20, 35 antioxidant properties in whole milk powder and in encapsulated fish
oil

and 50 °C for 4 months • The brown colour is beneficial in some foods, not dairy

Deeth & Lewis (2017)

Stages of the Maillard reaction

• The Maillard reaction is often considered to occur

in 3 broad stages: Early stage Maillard
• Early (lactosylation)
Reaction:
• Mid or advanced - formation of numerous products
• Late – formation of brown pigments
Lactosylation (glycation)

• Note: all stages can be occurring at the same time

 Lactosylation Lactosylation 2

• First stage of Maillard reaction in milk • Forms lactosyl-lysine (protein-bound)

• No brown colour produced in this stage • Lactosyl-lysine is unstable and spontaneously

converts to lactulosyl-lysine (by an Amadori
• Lactose reacts with the ε-NH2 group of lysine rearrangement)
• Lactulosyl-lysine (the Amadori product) is a stable
product

Lysine: the amino acid

Lysine in a protein

Lactosylation 3 Lactosylation 4

• Formation of lactulosyl-lysine makes lysine

nutritionally unavailable (lysine blockage)
• More than one lactose can react per protein
molecule
Amine group of lysine • Lactose molecules can react with different lysines
side-chain attached to • in mono- and di-lactosylated caseins, 7 of 14 lysines in
protein backbone αs1-casein and 5 of 11 lysines in β-casein were
Protein-bound lactosylated
lactulosylysine
(the Amadori product)
 Lactosylation 5 Lactosylation (measured as furosine)
increases during storage
• Initiated during processing but continues during
storage UHT milk

• Extent of lactosylation can be measured in 200

various ways but commonly as furosine 180 Indirect

160 processing

Furosine (mg/100 g protein)

• furosine is formed from lactulosyl-lysine by digestion 140

with acid 120

100

• yield of furosine from lactulosyl lysine is ~32% so 80

amount of blocked lysine can be estimated 60

Direct processing
40

• there is no furosine in milk 20

0
0 4 8 12 16 20 24
Storage time (weeks)
Elliott et al. (2005)

Trial with MPC80

Furosine change of MPC during storage
Samples kept for up to 12 weeks at:
3 humidities and
0 3 6 9 12
4 temperatures 44 66
1600

1200
Furosine (mg/100g protein)

800
Lactosylation measured as furosine
400

Colour measured by Minolta colourmeter – change in 84

0

colour ∆E was calculated from L*, a* & b* 1600

Temperature
1200 (°C)
25
800 30
35
400 40

0
0 3 6 9 12
Week
Panel variable: Humidity (%) Le et al. (2011a)
 Colour change (∆E) of MPC during Trial with 4 powders
storage for up to 12 weeks at:
3 humidities, 4 temperatures Samples of MPC80, WMP, SMP & WPC stored for up to
0 3 6 9 12 12 weeks at:
44 66

20

15 3 humidities and
10

5
4 temperatures
0
∆E

84

20

15
Temperature
(°C) Lactosylation measured as furosine
25
10 30
35
5 40

0
0 3 6 9 12
Week
Panel variable: Humidity (%)

A colour difference of ~ 3.5 can be noticed (Le et al. 2011a)

Lactosylation (measured as furosine) Effect of stabilisers/pH

increases during storage at 30°C
In powders: MPC80, WMP, SMP, WPC80
0 2 4 6 8 10 12 14 SHMP
In-container
3500
MPC80 44% RH WMP 3000
66% RH
84% RH 2500
sterilised 25% TSC
2000
reconstituted skim
Furosine (mg/100g protein)

1500
DSHP
1000
milk samples
500

3500
SMP WPC80
0 DHSP
3000

2500

2000 Control
1500

1000

500

TSC (trisodium citrate) & DSHP (disodium

0 2 4 6 8 10 12 14

Storage time (week)

Le et al. (2011b) hydrogen phosphate) increase pH Lewis, unpublished
 Blocked lysine: the result of the early
stage Maillard Reaction
• The lysine in lactulosyl-lysine is not available for
digestion
• The percentage of lysine blocked by the Maillard Mid/advanced stage Maillard
reaction can be determined from furosine analysis
Product % Blockage
Reaction
Raw milk 0
Pasteurised milk 0-2
UHT milk 0-10
In-container sterilised 10-15
Evaporated milk 15-20
Condensed milk 14-36
Spray-dried powder 0-7
Roller-dried powder 10-50
Infant formulae 5-34

Mehta & Deeth (2016)

Advanced Maillard Reaction Products Advanced Maillard Reaction Products

(AMRPs) (AMRPs) 2
• Includes many compounds
• Sometimes called Advanced Glycation End-Products
(AGEs) • All due to decomposition of lactulosyl-lysine
• Particularly in the medical field 1. Formic & acetic acids –
• Maillard reaction takes place in the human body as well as in – the reason for the decrease in pH of UHT milk during storage
food we eat
• Medical interest because some AGEs have been implicated in
some illnesses Stored
• May be pro-inflammatory but ‘jury is still out’ (Davis et al. 2016) UHT milk
• Discussion beyond scope of this presentation

Gaucher et al. (2008)

Advanced Maillard Reaction Products
(AMRPs) 3
3. Flavour compounds, e.g. aldehydes
– Major contributor to flavour of high-temperature treated
products, UHT milk, milk powders
– Lysyl-pyrraline, maltosine, maltol, β‐pyranone, 3‐ furanone,
cyclopentanone, galactosyl‐ isomaltol, acetylpyrrole,
pentosidine found in heated milk (van Boekel 1998)
4. HMF (hydroxymethyl furfural)
– Often used as index of heat treatment or Maillard reaction
– When measured, HMF (total) includes free HMF and HMF
formed from lactulosyl-lysine by reaction with oxalic acid
during analysis
– free HMF is much lower than total HMF
Late Stage Maillard
Reaction
More AMRPs
4. Dicarbonyl compounds:
– e.g. + several others
– Can cross-link proteins by acting as linking agents
• React with amine groups on adjacent protein molecules
- Cross-linking occurs during storage of long-life milk and
powders – forms high-molecular-weight proteins
- Role in cross-linking was demonstrated by incubation of a
reconstituted MPC powder with methyl glyoxal;
- it reproduced the same high-molecular-weight proteins seen in
stored powder and UHT milk (Le et al. 2013)
- Cross-linking can:
- stabilise the casein micelle and protect it in UHT milk
- Increase the viscosity of yogurt
- reduce the solubility of powders
Late stage products
• Formation of brown pigments, melanoidins
• Responsible for colour of many foods, e.g., coffee, cocoa,
honey, malt
• Molecular weights of 3,500 to 14,000 Da
• Have antioxidative and antimicrobial properties
• Do not appear to have adverse health effects
Wang et al. 2011
 Can we stop or inhibit the Maillard
Reaction
• Probably not but it can be slowed down in some
circumstances
Inhibition of Maillard • By adding reducing compounds such as cysteine
• By adding polyphenols such as those in tea, e.g., epicatechin
Reaction • Epicatechin (0.1%) added to milk before UHT processing
markedly reduced Maillard-derived flavour compounds and the
cooked flavour of the UHT milk
• At 0.2% added epicatechin, UHT milk tasted similar to
pasteurised milk
• Tea polyphenols also reduced browning in UHT milk as measured
by colour difference ∆E

(Colahan‐Sederstrom & Peterson 2005; Schamberger & Labuza 2007)

References
Colahan‐Sederstrom, P.M. & Peterson, D.G. (2005) Inhibition of key aroma compound generated during
ultrahigh‐temperature processing of bovine milk via epicatechin addition. Journal of Agricultural and Food
Chemistry 53(2), 398–402.
Davis, K., Prasad, C., Vijayagopal, P., Juma, S. & Imrhan, V. (2016) Advanced glycation end products,
inflammation, and chronic metabolic diseases: Links in a chain? Critical Reviews in Food Science and Nutrition
56, 989-998.
Deeth, H.C. and Lewis, M.J. (2017) High temperature processing of milk and milk products. Wiley Blackwell, Oxford
(ISBN: 978-1-118-46050-4) Publication date May 2017).

Thank you for your attention
Elliott, A.J., Datta, N., Amenu, B. and Deeth, H.C. (2005) Heat-induced and other chemical changes in commercial
UHT milks. J. Dairy Res. 72(4), 442-446
Gaucher, I., Molle, D., Gagnaire, V. & Gaucheron, F. (2008b) Effects of storage temperature on physico‐chemical
characteristics of semi‐skimmed UHT milk. Food Hydrocolloids 22, 130–143
Le, T. T., Bhandari, B. R., and Deeth, H. C. (2011a). Chemical and physical changes of milk protein concentrate
(MPC80) powder during storage. Journal of Agricultural and Food Chemistry, 59, 5465-5473.

Any questions? Le, T. T., Bhandari, B. R., Holland, J. and Deeth, H. C. (2011b). Maillard reaction and protein crosslinking in relation
to solubility of milk powders. Journal of Agricultural and Food Chemistry 59, 12473–12479
Le, T.T., Holland, J.W., Bhandari, B., Alewood, P.F. and Deeth, H.C. (2013) Direct evidence for the role of Maillard
reaction products in protein cross-linking in milk powder during storage. Int. Dairy J. 31, 83–91
Maillard, L.C. (1912). Formation of melanoidins in a methodical way. Compt. Rend.154, 66.
Mehta B.M. and Deeth H.C. (2016) Blocked lysine in dairy products – formation, occurrence, analysis and nutritional
implications. Comprehensive Reviews in Food Science and Food Safety 15, 206-218.
Schamberger, G.P. & Labuza, T.P. (2007) Effect of flavanoids on Maillard browning in UHT milk. LWT‐Food
Science and Technology 40, 1410–1417.
van Boekel, M.A.J.S.1998) Effect of heating on Maillard reactions in milk. Food Chemistry 62(4),403–414.
Wang, H.-Y., Qian, H. & Yao, W.-R. (2011) Melanoidins produced by the Maillard reaction: Structure and biological
activity. Food Chemistry 128, 573–584.