Food structure

vs. nutrient bioavailability

INFOGEST PhD Training School
Food Digestion and Human Health
Gdansk, Poland

23th of April 2013

Amlie Deglaire
amelie.deglaire@agrocampus-ouest.fr
 Introduction
Nutrient bioavailability
availability for the metabolic functions of the organism
nutrient content of a food
The best indicator of the nutritional quality of a nutrient
Influencing factors
Chemical state of the nutrient
Interactions with other food components
Presence of antinutritional factors
Release from the food matrix
Recent data : influence of the matrix state of natural foods or the
microstructure of processed foods ( or ) 1200

Number of scientific publications

1000

Increase of publications in food digestion 800

associating food scientists, nutritionists and 600

400

gut physiologists 200

0
1991 1995 2000 2005 2009
Year

How the food matrix structure influence food digestion?

2
 Outline
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
3
 Outline
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
4
 Food structure?
Food structure or food matrix = organisation of food constituents at multiple spatial
scales and their interactions
Provided by nature or imparted during processing and preparation (food manufacturing)
Impact on food quality in particular on sensory and nutritional aspects
Insight in food structure and how it changes during processing operations is essential for
producing high quality food

Multiple scale structural elements Interactions proteins - lipids

Mackie and Macierzanka, 2010

Source: Aguilera, 2005, 2007; COST Action FA1001: insidefood 5

 Example of food structures produced by milk
processing

Source : Aguilera, 2005 6

 Nutritional quality of a food

Food Nutrient content

Synthetic indicator : balance between the good and bad nutrients
Example: nutritional profil SAIN LIM (Darmon & Darmon, 2008)
Average % of the coverage or excess of the recommendations for
the nutrients of interest
SAIN : Score dAdquation Individuel aux recommandations Nutritionnelles /
Adequate Score for Nutritional Recommendations
Proteins, Fibres, Vitamine C, Calcium, Iron

LIM: score des nutriments LIMiter / Nutrients to limit

Sodium, Saturated Fatty Acids, Added Sugars

Recognized by the French authorities

7
 Impact of a process on the nutritionnal
quality profil
Apple drying

Source: Achir et al., Journal of Food Engineering, 2010 8

 Food digestion : regional specificity of the digestive tract

*
* No lingual lipase coded in humans.
Bacterial origin?

The physical properties of a food matrix can affect the efficiency of the
physical, enzymatic and chemical digestion processes

Source: Guerra et al., 2012, Trends in Biotechnology 9

 Nutritional quality of a nutrient

Nutrient bioavailability
Fraction of a nutrient that has been digested and absorbed
and is available for (has been used by) the metabolic functions of
the organism Metabolism

Blood
Urine

Absorption

Faeces

Raw Structured Mouth Stomach Duodenum Jejunum Ileum Colon

ingredients food

Digestion Excretion
10
 Use and development of in vivo models

Human adult Neonate

Adult Mini-pig Piglet

Bioavailability :
measure in the blood plasma
metabolic utilization : waste from digestion (faeces) + metabolism
(urine, blood) 11
 Bioavailability measure

Blood sampling
Bioavailability = Area Under the Curve (quantity of nutrient absorbed) x 100
quantity of nutrient ingested
12
 Bioavailability estimates
Metabolism

Blood
Urine

Absorption

Faeces

Raw Structured Mouth Stomach Duodenum Jejunum Ileum Colon

ingredients food

Digestion Excretion
Digestibility: (Ningested Nfaeces )/Ningested
Net Postprandial Protein Utilisation (NPPU):
(Ningested Nfaeces Nurine Nurea blood)/ Ningested
Biological value: NPPU/Digestibility

13
 Nutritional quality of a nutrient / in vitro

Nutrient bioaccessibility
fraction of a compound that is released from its matrix in the
gastro-intestinal tract during digestion and thus becomes
available for intestinal absorption (Fernandez-Garcia et al. , Nut Res, 2009)
Blood
Urine

Absorption

Faeces

Raw Structured Mouth Stomach Duodenum Jejunum Ileum Colon

ingredients food

Digestion Excretion
14
 Development of in vitro digestion models
Static Conditions Dynamic Conditions
Partnership with UMR GMPA Grignon

Gastric Duodenal Epithelial

Phase Phase Phase

Constant pH
No flow le VMBB : enzymes

Constant [Enzymes]
Regulated pH
Dynamic flow
Regulated[Enzymes]

Digestion models for the adult and the neonate

Bioaccessibility
15
 Multi-scale characterization of digested food

Proteins Lipids Microscopic scale

Stomach Duodenum Jejunum Ileum

Estomac
estomac Duodnum Jjunum Ilon
duodnum jjunum ilon
35 6.50

30 6.00
Will the liquid dairy sample Sample 1
25 5.50
coagulate when entering in
the stomach??? 20 5.00

G' (Pa)

pH
Prediction of the 15
pH
4.50

rheological behaviour of an 10 4.00

IF in gastric conditions 5 3.50

Sample 2
Viscosity of the gel
0 3.00
0 25 50 75 100 125 150 175 200

Macroscopic scale time (min)

16
 Multi-scale characterization of digested food
Antibody arrays Molecular scale

Mass Spectrometry ELISA -lactoglobulin

-lactalbumin
50%
Caseins
40%

30%

20%
More than 4700
10%
peptides identified in
Surprol the human jejunum 0%
Est 30' Est 1h30 Est 3h30

17
 Development of in vitro digestion & absorption models

CaCo-2 cells: human epithelial colorectal adenocarcinoma cells differentiated

enterocyte absorption
HT29-MTX cells : goblet cell clone mucus secretion (Walter et al., 2000)

Source : Prada & Aguilera, 2007 18

 Bioavailability & bioaccessibility

Source : Fernandez-Garcia et al. , Nut Res, 2009 19

 Outline
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
20
 Carbohydrate : starch
Starch microstructure: granular (2-130 m ) / cristalline

Amylose : linear structure

( gel)
Amylopectine : branched structure
( viscosity)

Amylose content varies with the botanic species (mean : 20% / waxy wheat : 0%)
Starch digestion hydrolysis of glycosidic bond
Linkage -1,4 : salivary and pancreatic amylase
Linkage -1,6 (amylopectine) : membrane isomaltase (after amylase)

21
 Starch granule organisation

Source Jenkins et al., 1996 22

Starch granule

Native and modified starch

granules 23
 Starch cooking
Raw starch : insoluble in cold water, low digestibility
Cooking with water and heat
gelatinisation : destruction of the cristalline structure, starch granule
swelling increased digestibility
retrogradation of starch (semi-cristalline structure) with cooling
decreased digestibility

Different behaviour
according to the botanical
sources (ex: potato vs. cereal)

Source: B. Boursier, techniques de lingnieur, 2005 24

 Structure evolution of starch during
gelatinisation

Light microscope
under polarised
light

Light microscope
under polarised
light

20 m

Source: Bogracheva et al. 2006 ; Crowther, 2012 25

 Starch cooking

Cooking with dry heat (dextrinisation)

Extrusion/milling :
starch destructuration: digestibility increased

Resistant starch (retrogradation, raw, non accessible,

high amylose content)
digestibility decreased( fonctionnal fibers )

26
Glycemic index : glucose release kinetics

Nature & structure

Food Carbohydrates -Composition
- amylose/amylopectine
Viscosity - associated nutrients
Energy -Mecanical and thermal treatments
Lipids Gastric - water cooking (glatinisation)
X emptying - extrusion (fusion)
-Dry cooking (dextrinisation)
-Milling

Digestion X
Absorption

27
 Glycemic index (GI)

1. 50 g of carbohydrates to test
2. Blood sampling (at regular intervals for 2-3 h)
3. Blood glucose content
4. Comparison with the reference (glucose)
5. Average on 8-10 volunteers

GI (%)=100 x AUC food

AUC reference

Source: Brand-Miller et al., AJCN, 2005 28

 Food GI
GI
Alone Mixed diet

High

Average

Low

29
 Food GI
GI
Alone Mixed diet

High

Average

Low

30
 GI & health

Glucose as reference
Low GI (spagetthis: 50) < 55
Average GI(pain blanc: 69) 55-70
High GI (corn flakes: 80) > 70

Low GI
prolonged glucose disposal
before exercise (toughness)
potentially with a satiating effect
hypoglycaemia delayed
High GI
short glucose disposal
after exercise: stock recovery
Diet with high GI : potential impact on weight gain, food intake,
triglyceride synthesis increased , oxydative stress
31
 Starch nature & GI
GI (Amylopectine) > GI(Amylose)
branched chain of amylopectin: more area accessible to enzymes
amylose: more rigid gel less accessible to enzymes/ more retrogradation

n=26

(Behall et al., Am J Clin Nutr 1988) 32

 Starch structure and digestibililty
In vitro digestion

DG85 Correlation among degree of

DG71 gelatinisation, digestion rate, plasma
glucose and insuline in rat
DG37

Metabolic response in rats

DG14

DG0

Starch gelatinisation increases its

digestibility

Degree of hydrolysis of starch at Source Holm et al, Am J Clin Nutr 1988

diffrent degree of gelatinisation (DG) 33
 Starch structure and GI

n=1

Starch cooking, degree of gelatinisation (DG) and blood

glucose (GI)

Gelatinisation of starch increases its glycemic index

Parada & Aguiliera, 2009,2012 34

 Starch structure and GI
Degree of gelatinisation
23 cereal products:
BC : breakfast cereals (5)
n=13
Degree of gelatinisation

BP&C : bakery products and crackers (6)

Bi : Biscuits (12)

Biscuits baked under very-low-moisture conditions

Degree of gelatinisation reduced

Glycemic index The higher the gelatinisation

degree, the higher is the GI
Impact of other nutrients

Source : Englyst et al. Br J Nutr 2003

35
 Degree of gelatinisation, flake thickness
and glycemic index
n=10

Product Kernel Thickness Gelatinisation GI

Pretreatment %
Bread (reference) 100a
Oat flakes Oven+Steam 0.5 mm 24 114 12a
Oat flakes Oven 0.5 mm 16 99 10ab
Oat flakes Steam 1 mm 16 76 8c
Oat flakes Oven 1 mm 16 72 9c
Oat flakes Raw 1 mm 0 78 9bc

Minimal processing of oat flakes : minor effect on GI features

compared with the more extensive commercial processing

Granfeldt et al, J Nutr 2000 36

 Process and composition impact
Common Bread Tradition Bread Sourdough bread

GI: 7511a D: 0.16 608a 0.24

538b 0.29
Common Bread + fibres Tradition Bread + fibres

GI and
texture
(density: D)
558b 0.30 Glycemic Insulinemic Bread
6213a 0.21 index index density

Dense Common Bread Dense Tradition Bread

538b 0.32
559b D: 0.24 Source: Rizkalla, 2009 37
 Resistant starch

Starch nature: amylose +++

RS1: native starch physically inacessible in entire crops (no grinding)

RS2: native starch granules that resist to digestion (no extensive

gelatinisation : moisture < 40 % ou T<Tgelatinisation)

RS3 : retrograded starch

High heat treatment (> 120C) favors the amylose cristallisation during cooling
and limits its capacity to re-gelification

RS4: starch chemically modified

38
Resistant starch & health

39
 Impact of a diet with high glycemic index
on health
n=3931 japanese women

Positive association between high GI diet, low fiber intake

& BMI
Source : Murakami et al., Eur J Clin Nutr 2007
40
 Impact of a diet with high glycemic index
on health

Cardiovascular disease risk increased by high GI diet

Mente et al., Arch Intern Med 2009

41
 Impact of a diet with a high glycemic
index on health

Increase of the oxidative stress in plasma

Plasma malondialdehyde (mol/L)

Plasma F2-isoprostane (ng/mL)

P for trend : p=0.03 P for trend : p=0.02

Quartiles of GI diet (Average GI: Q1,50.3; Q2, 53.6; Q3, 55.8; Q4, 59.9, n=291)
Adjusted data for age, gender, BMI, tobacco, alcool, kcal, proteins, fibres, folates and cholesterol

Source: Hu et al., Am J Clin Nutr 2006 42

 Impact of a diet with high glycemic index
on health
Diet with high GI increases the risk of non-insulinodependant
diabetes (type II)

Relative risk and confidence interval at 95% of diabete II according to the GI diet
quintile in 91 249 women
Source: Schulze et al., Am J Clin Nutr 2004 43
 Conclusion: Technology and glycemic
index
(1) Raw material choice (+++)

Balance amylose/amylopectine
Associated nutrients (proteins and fibres)

(2) Mecanical and thermal treatment

Water (gelatinisation) / Dry(dextrinisation)
Extrusion
Grinding

44
 Outline
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
45
Plasma 1200

1000

800

AA totaux (mol/L)
600

400
Am J Clin Nutr, 2006
200

0
0 1 2 3 4 5 6 7 8 9
-200

-400

n=23 Temps (h)

Prot. Sriques Casines

Kinetics of digestion influence kinetics of AA absorption

Small intestine Nombre de
peptides
90
Ntot (mmol/mL)
Caseins 80 Caseins
0,6 70 Whey Prot.
0,5 Whey Prot. 60
0,4 50
0,3 40
30
0,2
20
0,1
10
n=14 0
0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 5,5 6
0
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6
Time (h) Temps de digestion (h)

Surprol

46
Milk protein microstructure : casein micelle
Different theoretical models have been proposed

The submicellar model The hard-sphere model

Walstra & Jennes Holt 1994

1984

Dalgleish et Marchin et
al. al.
2004 2007
200 nm

The real organisation of individual caseins within the micelle remains unknown

47
Casein micelle reactivity
Alkalinisation Cooling
(micellar destructuration
-casein
H2O
Calcium
Calcium Caseins
&
phosphate Heattreatments
phosphate >90C
Acidification
Denaturated whey proteins
Protons
Calciumphosphate
Calciumphosphate

Casein(before precipitation) k-casein, peptides, NH3

H2O Lactose

Caseins

Cations

Anions
H2O
Addition of di H2O Casines
Addition
or trivalent Calciumphosphate
of
cations chelatants
Calcium Caseins H2O

Addition of NaCl

From F. Gaucheron, INRA

48
Milk protein microstructure : Whey proteins

Globular proteins

Known 3D-structures

-lactalbumin -lactoglobulin
Sensitive to heat-
denaturation

Highly resistant to
proteolysis (digestion)

Serumalbumin Lactoferrin

49
 Milk proteins

Casein micelle
MW = 0.5-1 x 106 kDa -lactoglobulin
av. diam = 180 nm MW = 18.6 kDa
2 SS bridges + 1 SH

-lactalbumin
k Casein
MW = 14.2 kDa
MW = 19.0 kDa
4 SS bridges
SS bridges

10
nm

Schematic representation of whey proteins and casein

micelle in a raw milk

50
 Milk macrostructure and N bioavailability

Gastric emptying
of PEG

Dietary N in plasma
In pig
Gaudichon et al,
J Nutr, 94

milk yogurt
Famelart et al., 2011

Intestinal fluxes
milk of dietary N

time
yogurt In human. Gaudichon et al, B J Nutr, 95

Dietary N absorption is slower for yogurt

than for milk
Kinetics of N bioavailibility related to
% N bioavailable
physical state of dairy products
(liquid/gel)

51
 Meat macrostructure
Energy needed to digest, absorb, and
assimilate meat meals in pythons

Source: Boback et al., Comparative Biochemistry and Physiology, 2007

Catheterised mini-pigs

Meat cubes
Minced meat

Kinetics of IAA bioavailibility

related to physical state of meat
(cube/minced)

52
 Meat macrostructure

Fast proteins (whey proteins)

Slow proteins (caseins)
Meat (high chewing efficiency)
Meat (low chewing efficiency)
Plasma leucine

Fast proteins favour postprandial

anabolism in elderly

Young adults Elderly

Anabolic utilisation
% of N ingested

NPPU
Fast Slow
Healthy Prothesis protein
protein
teeth

53
 Heat treatment & protein bioavailbility

Mild heat treatment (< 100C): protein conformation impacted

no impact or increase of overall digestibility

Source: Evenpoel et al., J Nutr, 1998

54
 Heat treatment of meat (<100) and bioavailability
of amino acids
Experiment conducted in mini-pigs (n=6) True ileal digestibility of N
60C: 94.7 0.5 %
75C: 96.3 0.4 %
95C: 95.1 0.7%

Cooking temperature (<100C) can modulate the speed of meat

protein digestion, without affecting the overall efficiency of the small
intestinal digestion
Source: Bax et al., Plos One, 2013 55
 Heat treatment & protein bioavailbility
High heat treatment (> 100C): modification of the primary
structure of the proteins
Formation of covalent bonds intra ou intermolecular (Lys, Arg) :
decrease of protein digestibility and amino acid bioavailability
Isomerisation of AA (LD): bioavailability decreased (Thr, Ile, Lys)
Maillard /Strecker reaction (reducing sugars): lysine bioavailability
reduced
AA destruction (Arg, Thr, Ser, Cys, Ile)
Toxic derivates (fire grill)
Example: formation of benzopyrene by pyrolysis of AA mutagen properties

56
 Raw 50C

Scanning Electron
Microscope
micrographs
60C 70C
Transverse sections
of beef muscle raw
and cooked

80C 90C

100C 121C

Source: Palka & Daun, Meat Science, 1999 57

 Meat cooking (100C) and protein digestibility
Effect of 100 C cooking on
protein hydrophobicity (Bromophenol Blue staining) as observed by microscopy (white scale bar: 100 m)

protein aggregation (Nile Red staining)

In vitro static model of digestion

gastric phase pancreatic phase
Addition of gastric and pancreactic digestion:
-quick cooking (100 C/5min or 270 C/1 min)
no effect on protein digestibility
-Long cooking (100C, 45 min)
decrease of digestibility (of 75%)

Source: Sant-Lhoutellier et al., J. Agric. Food Chem, 2008 58

 Gastric digestion & oxidative modification of
myofibrillar proteins

Proposed mechanism of pepsin action on meat proteins as raw or

heated at different cooking temperatures

Source: Sant-Lhoutellier et al., J. Agric. Food Chem, 2008; Bax et al, JAFC, 2012 59
Heat (&water) on in vitro digestibility of cereal protein
of pasta

Hydrolysis degree at 10 MIN

Low Temperature: 55C, High Temperature: 70, Very HT: 90C moisture level : 20%
VHT-LM: 90C - low moisture level : 12%

Source: Petitot et al., Food Chem, 2009 60

 Heat (&water) treatment impact on protein
aggregation
Elution profile of proteins by steric exclusion chromatography

Soluble proteins

Insoluble proteins

Source: Petitot et al., Food Chem, 2009 61

 Impact of milk heat treatment on casein digestion
CNs: major allergen in neonates
Native CNs very sensitive to hydrolysis

Sample preparation
Ultra low heat fat-free
milk powder

Step 1:
water suspension 25% Dry Matter 35% Dry Matter

Concentrate 1 Concentrate 2
Step 2 :
Heat 80C/20 s 105C/60 s 80C/20 s 105C/60 s
treatment
85C / 3 min 85C / 3 min

Step 3 :
Drying
control

In vitro digestion / neonate model

Source : Dupont et al. Mol Nutr Food Res, 2010 62

Impact of milk heat treatment on casein digestion
Neonate digestion : in vitro model

Milk powder Gastric Duodenal

phase phase

60 min 30 min
pH 3.0 pH 6.5
+ pepsin + trypsin
+ PC + chymotrypsin
+ bile salts

Sampling at 0, 1, 2, 5, 10, 20, 40 Sampling at 0, 1, 2, 5, 10, 15 and

and 60 min 30 min

Biochemical characterisation
SDS-PAGE, LC-MS-MS, ELISA

Source : Dupont et al. Mol Nutr Food Res, 2010

63
Impact of milk heat treatment on casein digestion
T
control
SDS-PAGE gels 66.3
of the digesta 55.4 0 1 2 5 10 20 40 60 1 5 15 30
36.5
31.0 Caseins
21.5
14.4

6.0
3.5
2.5
Gastric Duodenal

66.3 A
A: 25% DM, 80C, 20s B: 25% DM,B85C, 3 min C: 25% DM, C
105C, 60 s
55.4 66.3
55.4
36.5 0 1 2 5 10 20 40 60 1 5 15 30 0 1 2 5 10 20 40 60 1 5 15 30 0 1 2 5 10 20 40 60 1 5 15 30
31.0 36.5
21.5 31.0
14.4 21.5
14.4
6.0
3.5 6.0
2.5 3.5
2.5
Gastric Duodenal Gastric Duodenal Gastric Duodenal

E: 35% DM,E80C, 20s F: 35% DM, 85C,

F 3 min G: 35% DM,G105C, 60 s
66.3 66.3
55.4 55.4
0 1 2 5 10 20 40 60 1 5 15 30 0 1 2 5 10 20 40 60 1 5 15 30 0 1 2 5 10 20 40 60 1 5 15 30
36.5 36.5
31.0 31.0
21.5 21.5
14.4 14.4
6.0 6.0
3.5 3.5
2.5 2.5
Gastric Duodenal Gastric Duodenal Gastric Duodenal

Heat treatment (>100C) increases casein restistance to digestion

64
Peptide identification in digesta by LC-MS-MS

Peptide profile varies with process

Peptide Bioactivity A B C E F G T

-CN(f108-113) Anti-hypertensif

-CN(f114-119) Opiod agoniste

-CN(f193-202) Anti-hypertensif

-CN(f193-209) Immunomodulatoire

s1-CN(f1-23) Antimicrobien

s1-CN(f23-34) Anti-hypertensif

s1-CN(f91-100) Anti-stress

s1-CN(144-149) Antioxidant

-lg(f9-14) Anti-hypertensif
65
 Impact of milk structure on protein digestion and
amino acid bioavailability

Sample preparation
macrostructure
Ultra Low Heat unheated milk rennet gel
powder (raw milk) 24h-20 C,
pH 6.6
10 m
rehydration in rennet
water
heat treatment
90 C-10 min

heated milk rennet gel pH 6.6

24h-20 C,
rennet

microstructure

Source : Barb et al., 2013 66

Impact of milk structure on protein digestion and amino acid bioavailability

Impact of the microstructure (heat treatment) on protein digestion

unheated milk heated milk

caseins caseins
-lactoglobulin
-lactoglobulin

caseins: susceptibility to hydrolysis with or without heat treatment

-lactoglobulin: heat-induced susceptibility to hydrolysis
67
Heat-induced protein aggregation in milk
Micelle bound aggregates From Guyomarch & Famelart
casein
micelles
denatured -lg
denatured -la
micelle-bound
k casein
Jean et al., 2006 whey
aggregates
casein
10 nm s2

[a]
dissociation of
k casein

[b]
Casein micelle in heated milk, serum aggregates
in SEM (Harwalkar et al., 1989)

10 nm

Formation of soluble and micelle-bound aggregates

100 nm heat treatment (90 C, 10 min) =

Serum of heated milk
of whey protein denaturation
observed by SEM (Rodriguez and association to casein
del Angel & Dalgleish, 2006) micelles 68
 Impact of milk structure on amino acid
bioavailability n = 4 minipigs

Major impact of the macrostructure (liquid vs gel) on leucine bioavailibility

Smaller impact of the microstructure (heat treatment)
69
 Heat treatment impact on bioavailability of milk
proteins in rat

Caseins
Whey proteins
Unheated
Low-heat pasteurisation
High-heat p asteurisation
Utra High Temperature
Spray-dried milk

Lower metabolic utilisation of N for spray-dried milk

Source: Lacroix et al., JAFC, 2006 70

 Heat treatment impact on bioavailability of milk
proteins in human
80

Net postprandial protein

76

utilisation (%)
72

68

64
n=25
60
Microfiltered Pasteurised UHT

Dietary N incorporation into plasma amino acids

digestive kinetics modified

due to protein denaturation

Source: Lacroix et al, J Nutr, 2008 71

 Reverse chromatography (RP-HPLC) of milk proteins
Type of milk

Unheated

Low-heat pasteurisation

High-heat p asteurisation

Utra High Temperature

Spray-dried milk

Caseins

Whey proteins

Source: Lacroix et al., JAFC, 2006 72 72

 Reverse chromatography of whey proteins

Unheated milk

Low-heat pasteurisation

High-heat p asteurisation
OD 214 nm

Utra High Temperature

Protein aggregation
+ Lactosylation of
Spray-dried milk
beta-lactoglobulines :
first products of the
Maillard reaction Caseins

Whey proteins

0 10
Time (min) Source: Lacroix et al., JAFC, 2006 73
Technological treatment and protein bioavailability

Alkaline treatment
Vegetal protein extraction
Detoxification
Ex: destruction of aflatoxines
Solubilisation and texturation of vegetal proteins
Destruction/modification AA :
Isomerisation of AA (Thr, Ile, Lys) from L to D
Neoformation of AA (cys et phosphosrine
dehydrolalanine) / formation of covalent bond type :
lysinoalanine (heat treatment)
Reduction of protein digestibility
Reduction of lysine bioavailability

74
 Metabolic utilisation reduced for some D-amino acids

Source: Friedman et al., JAFC, 1999 75

Protein interaction with other nutrients

Proteins - lipids
Hydrophobic interaction
Covalent bond with lipid oxidation derivate
Reduction of lysine bioavailability and protein digestibility
Proteins-polyphenols (vegetal food)
Covalent bond between lysine and quinone (polyphenol
oxidation by enzymes or alkaline pH)
Reduction of lysine bioavailability and protein digestibility
Proteins - nitrites, sulfites or chlore derivates
Minimal decrease of bioavailability
Formation of toxic compounds (nitrosamines, dichlorovinylcystine,
methionine sulfoximine)

76
 Outline
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids
3. Food structure and micronutrient bioavailability
1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
77
 Technological treatment and lipid bioavailability

Partial hydrogenation (less sensitive to oxidation, liquid solid)

Insaturated fatty acid cis trans

Example : margarine (1-2 % for new process, 15 up to 60 % for poor quality process) (ANSES, 2005)
Metabolised instead of other insaturated fatty acids
Increased risk of cardiovascular diseases for a daily intake of trans-
fatty acids > 2 % of the total daily energy intake
increase of LDL cholesterol and decrease of HDL cholesterol
Insaturated satured (partially) fatty acids
Increased risk of cardiovascular diseases

78
Saturated fatty acids and cardiovascular disease risk

protective

At risk

Source: deOliveira et
al., AJCN, 2012

79
 Technological treatment and lipid bioavailability

High heat treatment (frying)

Destruction of indispensable fatty acids
Toxic compounds
Lung cancer in women inhaling vapor from fish frying in China & Taiwan
(Yang et al., 2000)
Lipid peroxdation : malondialdehyde (MDA) released with prolonged
frying mutagen effect of MDA but not of the oil (Saguy & Dana, J Food
Engineer, 2003)
Maillard reaction in meat: heterocyclic amines
(reduced release for T<200C, spices (antioxydant), microowaves)
risk factor for colon cancer
Trans fatty acids (repeated frying/ exchange food-oil)
Hydrolysis of ester bond free fatty acids more oxidable
Oxidation hydroperoxide plasma oxidative stress

80
 Impact of fat droplet size on lipid bioavailability
Human experiment (n=8)
Emulsion : 50g olive/fish oil (+ carbohydrates + protein)
Fat droplet size : Fine emulsion : 10 m vs. Coarse emulsion : 0.7 m

Stomach lipid digestibility

7-16% vs 20-37%

Duodenal lipid digestibility

37-46% vs 57-73%

Source: Armand et al., AJCN, 1999 81

 Impact of the emulsifying agent on lipid digestion (in vitro)
Tween 20 Lysolect. Caseinate Whey Prot

Before digestion

+ saliva 5 min

+ gastric juice 2 h

+ dudodenal juice
2h
Source: Hur et al., Food Chem, 2009 82
 Impact of the emulsifying agent on lipid bioavailability

stearoyl-lactilate sodium

Postprandial triglycerids in the plasma

(n=8 healthy subjets)

Source: Golding et al., Soft Matter, 2011 83

 Impact of fat structure on bioavailability
Spread (non-emulsified)

Emulsion (oil-in-water/milk)

n=9 n=9
Fat structure impacts fatty
acid and chylomicron plasma
kinetics but not overall
digestibility
Source: Vors et al., AJCN, 20123 84
 Impact of fat structure on metabolic oritentation

Fat structure impacts fatty

acid oxidation kinetics

Slow vs fast fat

Fat structure impacts digestive More oxidation for rapidly

kinetics and subsequently metabolic digested fat (emulsifion) than
orientation for non-emulsified fat

Source: Vors et al., AJCN, 20123 85

 Blood triglycerides and cardiovacular
disease risk

Source: Michalski, Eur. J. Lipid Sci, 2009 86

Human milk pasteurisation : impact on protein & lipid
digestion
KDa
IgA
200 IgG
116 X0
97
66 Lf
SA
55
Bt

36
31 Cas
Cas
21
14
-lac

6
3.5
2.5

0 30 60 120 150
Time (min)

Gastric digestion
Pasteurisation
Aggregation of proteins
around the native fat globule

Mild heat treatment in human milk

Proteolysis increased for caseins and lactoferrin
No impact on whey proteins (SA and alpha-lac)

87
 Human milk pasteurisation : impact on protein & lipid
digestion
0 min
30 min
60 min
120 min
150 min

Triglycerids

Free Fatty Acids

Diglycerid s (sn-1,3) /
Diglycerid s(sn-1,2) Cholesterol

Monoglycerid
PL/Prot
Time 0 30 60 120 30 60 120 180
(min)

Gastric digestion Intestinal digestion

Lipolysis reduced in the gastric digestion in

Grey intensity

75
50 pasteurised milk vs. raw milk
(AU)

25
Due to BSSL inactivation and structure effect ?
0
0 30 60 120
88
 Outline
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
89
 Micronutrients from plant foods and location
within the tissue

Source: Parada & Aguilera, 2007

90
 Plant processing and micronutrient bioavailability

Source: Parada & Aguilera, 2007 91

 Outline
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
92
 Carotenoids
Carotenoids : natural pigments yellow-orange red-purple,
highly hydrophobic compounds located in specialized plant
plastids (chromoplasts) of fruits, vegetables, mushrooms, algae
About 10 % of carotenoids (bta-, alpha-, gamma-carotne,
cryptoxanthine): vitamin A precursor
Beta-carotne: orange fruits (carott, apricot, mango), red palm oil,
dark green vegetables (hidden by chlorophylle)
Vitamin A: necessary for vision, epithelium tissu preservation,
bone growth, immune system

Liver Retine Bone, mucosa

(storage form) (Vision)

93 93
 Carotenoids
Absorption: requires incorporation of lipophilic
carotenoids into mixed micelles (dietary lipids, bile
salts, and other lipophilic or amphiphilic compounds)
Liberation : fraction of the test compound released from the food matrix
into the aqueous supernatant
Bioaccessibility : fraction of the test compound released from the food
matrix into the aqueous supernatant and transferred into mixed micelles
Bioaccessibility / Bioavailability vary according to the
botanical origin : Green vegetables < oranges vegetables <fruits < oil
Process (thermal and/or mecanical) : increase the
bioavailability of carotens by destruction of the
vegetal matrix (Van Buggenhout et al., Trends in Food Science & Technology 2010)

94
 Chromoplast structure & bioaccessibility of
carotenoids
BA: 0.5%d BA: 3.1%c
Beta-carotene: 13 080 g/100g Beta-carotene: 652 g/100g

Crystalloid

BA: 10.1%a BA: 5.3%b

Beta-carotene: 680 g/100g Beta-carotene: 485 g/100g

Globular-tubular

Light micrographs of carrot root and mango, papaya, and tomato fruit
mesocarp. Arrows mark chromoplasts.

Source: Schweiggert et al, Food Chem, 2012 95

 Chromoplast structure & bioaccessibility of
carotenoids
Bioaccessibility & oil addition
Liberation and bioaccessibility

Impact of
pectin and
other fibers

Source: Schweiggert et al, Food Chem, 2012 96

Carott processing and bioavailability of carotenoids

Carotts: cooking increases bioavailability of beta-carotenes

Plasma beta-carotene : 2-3 times higher for cooked and process
carotts (canning and heating at 121 C)

Source: Rock et al., 1998; Livny et al., 2003 97 97

Carott processing and bioavailability of carotenoids
Carotts: cooking increases bioavailability of beta-carotenes
Plasma beta-carotene for commercial puree carrot > home-made
puree carrot and raw grated carrot (Edwards et al., 2002)

Vitamin A conversion
Commercial puree: 44 11%a
Home-made puree: 59 12%b
Grated carrot: 63 10%ab

Research still necessary to optimise process maximizing

carotenoid bioavailability

98 98
 Addition of oil into carotts and bioavailability
of carotenoids

Effect of addition of oil on micellarisation of carotenoids from crude

and cooked carrots in respect to the carotenoid content in the corresponding
digest.

Hornero-Mendez et al., 2007 99

 Plan
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
100
 Iron & spinach

Breakdown of
cellular
structures

Digestion of
cellular
structures

Source : Crispin et al., 2002 101 101

 Plan
Plan
1. Definition
1. Food structure
2. Bioavailability vs. bioaccessibility

2. Food structure and macronutrient bioavailability

1. Carbohydrates
2. Proteins
3. Lipids

3. Food structure and micronutrient bioavailability

1. Micronutrients from plant food
2. Carotenoids and beta-carotenes
3. Iron

4. Conclusion
102
 Improving health properties of food by
sharing our knowledge on the digestive
process
COST Action FA1005
INFOGEST

Chair: Dr. Didier DUPONT, Senior Scientist, INRA, France

ALIMENTATION
AGRICULTURE
ENVIRONNEMENT

Beneficial food component characterization
Stability of bioactives during processing
Multi-scale characterization of food
Harmonization of in vitro digestion models
Comparison in vitro / in vivo
Characterization of the digestion products
Resistance to peptidases and absorption
Active form of food components in the organism

Characterization of raw In vitro, in vivo and in

materials and processed silico models of
food matrices for optimized mammalian
nutrient bioaccessibility gastrointestinal digestion
WG1 WG2
Dairy
Fruits & Vegetables
Egg

Evaluation of the health

effects
WG3

- Food allergenicity and immunomodulatory properties

- Kinetics of digestion / regulation of appetite and satiety
- Protein metabolization on human microbiota
- Impact of ACE inhibitors on cardiovascular diseases

104
 Tech Univ Denmark Univ Aarhus MTT Univ Eastern Finland
Canada
Norwegian Univ Life Sci VTT Nofima
Wageningen UR Rothamsted Res

Teagasc Univ Reading Univ Leeds

Cent Rech Lippmann Laval Univ
Univ Ghent
Agroscope Posieux Univ Guelph
Inst Food Res

Leatherhead Food Res KTU Food Inst

Polish Academy of Sci

Univ Greifswald
Univ Belgrade
Max Rubner-Institut
INRA Univ Novi Sad

AgroParisTech Centr Food Res Inst

CSIC Ben Gurion Univ

Technion
Univ Alto Douro
ITQB
Milan State Univ Pom Med Univ New Zealand
Ege Univ
Univ Bologna

150 scientists - 50 institutions 26 countriesRiddet Inst

105
 The application of innovative
fundamental food-structure-property
relationships to the design of foods for
health, wellness and pleasure
COST Action FA1001
Food Structure Design

http://www.insidefood.eu

106
 Food structure vs. bioavailability

Source: Parada & Aguilera, 2007

107
 Conclusion
Nutritionnal quality of a food not only impacted by
nutrient content but also by its structure
Key factor for processed foods
Food structure impacts on digestive events (gastric
phase: key step) and subsequently on metabolic
orientation
Food structure : key parameter to control or modify
nutrient bioavailability nutrition for specific
popultation (elederly, sportmen, baby..)
Relationship among food structure/digestion/health
burning topic in science
further knowledge requiredALIMENTATION
AGRICULTURE
ENVIRONNEMENT