Imre Blank, Philippe Pollien, Chahan Yeretzian
Imre Blank, Philippe Pollien, Chahan Yeretzian
Imre Blank, Philippe Pollien, Chahan Yeretzian
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<strong>Imre</strong> <strong>Blank</strong>, <strong>Philippe</strong> <strong>Pollien</strong>, <strong>Chahan</strong> <strong>Yeretzian</strong><br />
Nestlé Research Centre<br />
Lausanne, Switzerland<br />
PTR-MS Igls / IBk January 2003 1
Outline of the presentation<br />
Basic principles of the Maillard reaction<br />
Major reaction phases, intermediates, its role in food and medicine<br />
PTR-MS data for reaction mechanisms<br />
Maillard model systems based on proline<br />
Main approaches in aroma research<br />
Sensory directed chemical analysis<br />
Formation kinetics of key odorants by PTR-MS<br />
Maillard model systems based on proline<br />
… but will not go into technical details of PTR-MS<br />
PTR-MS Igls / IBk January 2003 2
Importance of the Maillard reaction<br />
00-now<br />
95-99<br />
Time period (year)<br />
90-94<br />
85-89<br />
80-84<br />
75-79<br />
70-74<br />
65-69<br />
Food<br />
Flavours<br />
Medicine<br />
MS<br />
0 200 400 600 800 1000 1200 1400<br />
Publication number<br />
PTR-MS Igls / IBk January 2003 3
Aldose<br />
+ amino compound<br />
-H 2<br />
O<br />
A<br />
N-substituted glycosylamine<br />
B<br />
Amadori rearrangement<br />
C<br />
1-Amino-1-deoxy-2-ketose<br />
(Amadori compound)<br />
D<br />
Schiff base<br />
of HMF<br />
or furfural<br />
+ H 2<br />
O<br />
+ amino<br />
compound<br />
-3 H 2<br />
O<br />
- amino<br />
compound<br />
HMF<br />
or furfural<br />
G<br />
F<br />
Aldimines<br />
E<br />
-CO 2<br />
Aldehyde<br />
+ amino acid<br />
Strecker<br />
degradation<br />
+ amino<br />
compound<br />
Aldoles and<br />
N-free<br />
polymers<br />
+ amino<br />
compound<br />
C<br />
-2 H 2<br />
O<br />
Reductones<br />
-2H +2H<br />
dehydroreductones<br />
G G G<br />
F<br />
Fission<br />
products<br />
(acetol,<br />
pyruvaldehyde,<br />
diacetyl etc.)<br />
F<br />
+ amino<br />
compound<br />
G<br />
Aldimines or<br />
ketimines<br />
G<br />
(Hodge, 1953)<br />
Aroma<br />
Taste<br />
Colour<br />
Antioxidants<br />
Texture<br />
Nutritional value<br />
Contaminants<br />
Toxic compounds<br />
MELANOIDINS (brown nitrogenous polymers and copolymers)<br />
PTR-MS Igls / IBk January 2003 4
Major fluxes in the Maillard reaction<br />
HO<br />
HO<br />
OH<br />
O<br />
OH<br />
R<br />
N<br />
H<br />
N-Glycosyl<br />
COOH<br />
OH<br />
HO<br />
OH<br />
R<br />
+<br />
HO<br />
H 2<br />
N COOH<br />
O OH<br />
H 2<br />
O<br />
Aldose sugar<br />
Amino acid<br />
HO<br />
HO<br />
OH<br />
OH<br />
OH<br />
R<br />
N<br />
H<br />
COOH<br />
N H 2<br />
A<br />
R<br />
COOH<br />
HO<br />
HO<br />
OH<br />
3-DO<br />
O<br />
O<br />
O H 2<br />
HO<br />
HCOOH<br />
O<br />
HMF<br />
CHO<br />
A : 1,2-enolisation<br />
(pH < 7)<br />
OH<br />
Flavour<br />
precursor<br />
Formation of volatile compounds in prolinebased<br />
Maillard reaction systems<br />
Reaction conditions<br />
Precursors<br />
Glucose/L-proline (1:1), Fru-Pro (0.2 mol)<br />
Buffer Phosphate (50 mL, 0.2 M, pH 5-8)<br />
Temperature<br />
Time<br />
Analytical methods<br />
Boiling (reflux)<br />
Up to 7 h<br />
PTR-MS: Direct headspace analysis<br />
GC-MS: Tenax trapping, solvent extraction<br />
PTR-MS Igls / IBk January 2003 6
Tenax<br />
Experimental set-up<br />
for Maillard PTR-MS<br />
HS Conc [ppbv]<br />
2000<br />
1500<br />
1000<br />
500<br />
0<br />
mass 45 mass 47 mass 51<br />
mass 57 mass 59 mass 71<br />
mass 75 mass 87 mass 145<br />
mass 43 mass 61<br />
0 50 100 150 200 250 300<br />
Time[min]<br />
8000<br />
7000<br />
6000<br />
5000<br />
4000<br />
3000<br />
2000<br />
1000<br />
0<br />
HS Conc. [ppbv], masses 43 and 61<br />
(T= 90°C)<br />
On-line monitoring of volatiles<br />
generated upon heating<br />
PTR-MS Igls / IBk January 2003 7
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
OH<br />
O<br />
OH<br />
OH<br />
OH<br />
OH<br />
OH<br />
R<br />
N<br />
H<br />
OH<br />
R<br />
N<br />
H<br />
O<br />
N<br />
H<br />
R<br />
COOH<br />
COOH<br />
COOH<br />
Amadori compound<br />
HO<br />
OH<br />
OH<br />
OH<br />
R<br />
N<br />
H<br />
COOH<br />
N H 2<br />
N H 2<br />
OH<br />
HO<br />
OH<br />
+<br />
HO<br />
O OH<br />
H 2<br />
O<br />
A<br />
R<br />
B<br />
R<br />
COOH<br />
COOH<br />
HO<br />
HO<br />
Hexose sugar<br />
HO<br />
HO<br />
OH<br />
3-DO<br />
O<br />
OH<br />
O<br />
O<br />
Melanoidins<br />
Flavour<br />
1-DO<br />
O<br />
[H]<br />
O H 2<br />
O H 2<br />
HO<br />
N H 2<br />
O<br />
HO<br />
R<br />
O<br />
O<br />
O<br />
COOH<br />
Amino acid<br />
HCOOH<br />
HMF<br />
O<br />
HDMF<br />
OH<br />
CH 3 OOH<br />
DDMP<br />
CHO<br />
OH<br />
HS Conc[ppbv]<br />
HS conc. [ppbv]<br />
180<br />
160<br />
140<br />
120<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
H<br />
?<br />
O<br />
OH<br />
m/z = 47 Formic acid + X<br />
0 50 100 150 200 250 300<br />
Glu/Pro<br />
Amadori<br />
8000<br />
7000<br />
6000<br />
5000<br />
4000<br />
3000<br />
2000<br />
1000<br />
0<br />
O<br />
OH<br />
Time[min]<br />
m/z = 61 Acetic acid<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
PTR-MS Igls / IBk January 2003 8
Coupling of GC with EI-MS and PTR-MS<br />
PTR-MS Igls / IBk January 2003 9
HS Conc[ppbv]<br />
180<br />
160<br />
140<br />
120<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
m/z = 47 Formic acid + X<br />
0 50 100 150 200 250 300<br />
Time[min]<br />
HS conc. [ppbv]<br />
8000<br />
7000<br />
6000<br />
5000<br />
4000<br />
3000<br />
2000<br />
1000<br />
0<br />
m/z = 61 Acetic acid<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
RT First trap Second trap<br />
(min) Peak hight % Peak hight %<br />
4.46 7400 75 3847 34<br />
12.5 2419 25 2234 19<br />
33.45 0 0 5399 47<br />
0 0 0 0<br />
Total 9819 100 11480 100<br />
RT First trap Second trap<br />
(min) Peak hight % Peak hight %<br />
10.5 6165 4 0 0<br />
31.2 136646 96 7470083 100<br />
0 0 0 0<br />
0 0 0 0<br />
Total 142811 100 7470083 100<br />
1st trap<br />
2nd trap<br />
: 10-15 min<br />
: 1h25-1h30<br />
PTR-MS Igls / IBk January 2003 10
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
OH<br />
O<br />
OH<br />
R<br />
N<br />
H<br />
COOH<br />
Amadori compound<br />
HO<br />
OH<br />
OH<br />
OH<br />
OH<br />
OH<br />
OH<br />
OH<br />
R<br />
N<br />
H<br />
O<br />
N<br />
H<br />
R<br />
OH<br />
R<br />
N<br />
H<br />
COOH<br />
COOH<br />
COOH<br />
N H 2<br />
N H 2<br />
OH<br />
HO<br />
OH<br />
+<br />
HO<br />
O OH<br />
H 2<br />
O<br />
A<br />
R<br />
B<br />
R<br />
COOH<br />
COOH<br />
HO<br />
HO<br />
Hexose sugar<br />
HO<br />
HO<br />
OH<br />
3-DO<br />
O<br />
OH<br />
O<br />
O<br />
Melanoidins<br />
Flavour<br />
1-DO<br />
O<br />
[H]<br />
O H 2<br />
O H 2<br />
HO<br />
N H 2<br />
HO<br />
O<br />
HO<br />
R<br />
O<br />
O<br />
COOH<br />
Amino acid<br />
HCOOH<br />
O<br />
O<br />
O<br />
HMF<br />
HDMF<br />
OH<br />
CH 3 OOH<br />
DDMP<br />
OH<br />
CHO<br />
HS conc. [ppbv]<br />
HS conc. [ppbv]<br />
12<br />
9<br />
6<br />
3<br />
0<br />
HO<br />
O<br />
m/z = 127 HMF<br />
CHO<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
Glu/Pro<br />
Amadori<br />
(pH 7)<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
m/z= 145 3,5-Dihydroxy-2,3-dihydro-6-methyl-4H -<br />
pyran-4-one<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
HO<br />
O<br />
O<br />
OH<br />
PTR-MS Igls / IBk January 2003 11
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
HO<br />
OH<br />
O<br />
OH<br />
OH<br />
OH<br />
OH<br />
OH<br />
R<br />
N<br />
H<br />
OH<br />
R<br />
N<br />
H<br />
O<br />
N<br />
H<br />
R<br />
COOH<br />
COOH<br />
COOH<br />
Amadori compound<br />
HO<br />
OH<br />
OH<br />
OH<br />
R<br />
N<br />
H<br />
COOH<br />
N H 2<br />
N H 2<br />
OH<br />
HO<br />
OH<br />
+<br />
HO<br />
O OH<br />
H 2<br />
O<br />
A<br />
R<br />
B<br />
R<br />
COOH<br />
COOH<br />
HO<br />
HO<br />
Hexose sugar<br />
HO<br />
HO<br />
OH<br />
3-DO<br />
Sugar<br />
O<br />
OH<br />
O<br />
O<br />
fragmentation<br />
1-DO<br />
O<br />
[H]<br />
O H 2<br />
O H 2<br />
HO<br />
N H 2<br />
HO<br />
O<br />
HO<br />
R<br />
HCOOH<br />
O<br />
O<br />
O<br />
COOH<br />
Amino acid<br />
O<br />
O<br />
HMF<br />
HDMF<br />
OH<br />
CH 3 OOH<br />
DDMP<br />
OH<br />
CHO<br />
HS conc. [ppbv]<br />
HS conc. [ppbv]<br />
600<br />
500<br />
400<br />
300<br />
200<br />
100<br />
0<br />
O<br />
m/z = 57 1-Hydroxy-2-propanone<br />
OH<br />
0 50 100 150 200 250 300 350<br />
Glu/Pro<br />
Amadori<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
Reaction time [min]<br />
m/z = 89 3-Hydroxy-2-butanone<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
(pH 7)<br />
O<br />
OH<br />
PTR-MS Igls / IBk January 2003 12
Principle approaches in aroma research<br />
OAV =<br />
C x<br />
T x<br />
C x : Concentration<br />
T x : Threshold value<br />
2<br />
SH<br />
SH<br />
O<br />
SH<br />
OH<br />
1<br />
OH<br />
SH<br />
SH<br />
O<br />
O<br />
OH<br />
Concentration (µg/L)<br />
1000<br />
800<br />
600<br />
400<br />
200<br />
0<br />
Sauvignon<br />
blanc<br />
1 2 3 4 5<br />
25<br />
20<br />
15<br />
10<br />
PTR-MS Igls / IBk January 2003 13<br />
5<br />
0<br />
Odor Activity Value
Formation of odorants in proline-based<br />
Maillard reaction systems<br />
Reaction conditions<br />
Precursors<br />
Glucose/L-proline (1:1), Fru-Pro (0.2 mol)<br />
Buffer Phosphate (50 mL, 0.2 M, pH 5-8)<br />
Temperature<br />
Time<br />
Analytical methods<br />
Boiling (reflux)<br />
Up to 7 h<br />
GC-O, GC-MS: Solvent extraction<br />
PTR-MS: Direct headspace analysis<br />
PTR-MS Igls / IBk January 2003 14
Volatile compounds formed in proline-based<br />
Maillard model systems 1<br />
Fru-Pro: Caramel-like, roasty Glc/Pro: Roasty, popcorn-like<br />
O<br />
O<br />
OH<br />
O<br />
OH<br />
O<br />
OH<br />
O<br />
OH<br />
O<br />
O<br />
<br />
HO<br />
O<br />
OH<br />
CH HO 3 COOH HCOOH<br />
O<br />
OH<br />
HO<br />
O<br />
OH<br />
OH<br />
O<br />
OH<br />
O<br />
OH<br />
<br />
O<br />
<br />
O<br />
O<br />
O<br />
<br />
N<br />
O<br />
N<br />
H<br />
O<br />
N<br />
N<br />
<br />
O<br />
O<br />
N<br />
H<br />
O<br />
PTR-MS Igls / IBk January 2003 15
Formation of key odorants in Maillard systems<br />
based on proline 2<br />
Concentration (ug/mmol)<br />
Concentration (ug/mmol)<br />
25000<br />
20000<br />
15000<br />
10000<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
5000<br />
0<br />
OH<br />
0 1 2 3 4 5<br />
Time (h)<br />
N<br />
H<br />
0 1 2 3 4 5<br />
Time (h)<br />
O<br />
N<br />
O<br />
O<br />
K<br />
Concentration (ug/mmol)<br />
Amadori<br />
Glu/Pro<br />
(pH 7)<br />
Concentration (ug/mmol)<br />
300<br />
250<br />
200<br />
150<br />
100<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
50<br />
0<br />
0 1 2 3 4 5<br />
Time (h)<br />
PTR-MS Igls / IBk January 2003 16<br />
O<br />
O<br />
OH<br />
0 1 2 3 4 5<br />
Time (h)<br />
N<br />
O
Monitoring of key odorants by PTR-MS<br />
HS conc. [ppbv]<br />
HS conc. [ppbv]<br />
8000<br />
7000<br />
6000<br />
5000<br />
4000<br />
3000<br />
2000<br />
1000<br />
0<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
O<br />
O<br />
O<br />
OH<br />
OH<br />
m/z = 61 Acetic acid<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
m/z = 129 Furaneol<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
HS conc. [ppbv]<br />
Glu/Pro<br />
Amadori<br />
(pH 7)<br />
HS conc. [ppbv]<br />
1000<br />
900<br />
800<br />
700<br />
600<br />
500<br />
400<br />
300<br />
200<br />
100<br />
0<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
m/z= 87 Diacetyl<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
m/z = 126 2-Acetyltetrahydropyridine<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
N<br />
O<br />
O<br />
O<br />
PTR-MS Igls / IBk January 2003 17
Glucose/proline (90°C)<br />
Tenax trapping (1h20 to 1h40min)<br />
GC EI-MS/PTR-MS coupling<br />
O<br />
PTR-MS Igls / IBk January 2003 18<br />
Acetaldehyde<br />
3-Hydroxy-2-butanone<br />
1-Hydroxy-2-propanone<br />
Acetic acid<br />
2-Acetyltetrahydropyridine<br />
O<br />
O<br />
N<br />
Acetone<br />
Diacetyl
Key odorants below the detection limit of PTR-MS<br />
(sample 50-fold diluted)<br />
3.5<br />
m/z = 112 2-Acetyl-1-pyrroline<br />
12<br />
m/z = 127 3,4-Dimethylcyclopentenolone<br />
HS conc. [ppbv]<br />
3<br />
2.5<br />
2<br />
1.5<br />
1<br />
0.5<br />
0<br />
Reaction time [min]<br />
0 50 100 150 200 250 300 350<br />
HS conc. [ppbv]<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
Reaction time [min]<br />
0 50 100 150 200 250 300 350<br />
Glu/Pro<br />
Amadori<br />
(pH 7)<br />
N<br />
O<br />
O<br />
OH<br />
PTR-MS Igls / IBk January 2003 19
H O<br />
H O<br />
O<br />
OH<br />
O<br />
OH<br />
OH<br />
OH<br />
+<br />
N<br />
N<br />
H<br />
O<br />
COOH<br />
O<br />
HS conc. [ppbv]<br />
600<br />
500<br />
400<br />
300<br />
200<br />
100<br />
0<br />
m/z = 57 1-Hydroxy-2-propanone<br />
(pH 7)<br />
Glc/Pro<br />
Amadori<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
N<br />
N<br />
H<br />
O<br />
N<br />
OH<br />
H<br />
N<br />
N<br />
H O<br />
O<br />
(Schieberle et al., 2000)<br />
O<br />
O<br />
[O]<br />
H 2<br />
O<br />
CO 2<br />
O<br />
HS conc. [ppbv]<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
m/z=126 2-Acetyl-1,4,5,6-tetrahydropyridine<br />
(pH 7)<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
Glc/Pro<br />
Amadori<br />
PTR-MS Igls / IBk January 2003 20
Good correlation of PTR-MS and GC-IDA data<br />
HS conc. [ppbv]<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
m/z = 126, Acatyltetrahydropyridine<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
Glu/Pro<br />
Amadori<br />
HS conc. [ppbv]<br />
PTR-MS<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
Glu/Pro<br />
Amadori<br />
m/z = 129, Furaneol<br />
0 50 100 150 200 250 300 350<br />
Reaction time [min]<br />
Concentration (ug/mmol)<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
0 1 2 3 4 5 6<br />
Time (h)<br />
K<br />
Glu/Pro<br />
Amadori<br />
Concentration (ug/mmol)<br />
GC-IDA<br />
300<br />
250<br />
200<br />
150<br />
100<br />
50<br />
0<br />
0 1 2 3 4 5 6<br />
Time (h)<br />
K<br />
Glu/Pro<br />
Amadori<br />
PTR-MS Igls / IBk January 2003 21
Conclusions<br />
Key odorants can be monitored by PTR-MS<br />
Furaneol, acetic acid, diacetyl, 2-AP, ATHP, methional, etc.<br />
Good correlation between PTR-MS and GC-IDA data<br />
Comparable kinetic curves<br />
PTR-MS gives complementary information to chromatography-based tools<br />
rapid, time-resolved, dynamic changes of volatiles, mechanistic insight<br />
Broad application in thermal-induced reactions<br />
Maillard reaction, lipid oxidation, flavour formation, off-flavours, ...<br />
Some remaining issues:<br />
Sensitivity, fragmentation, quantification, selectivity, ...<br />
PTR-MS Igls / IBk January 2003 22
Acknowledgements<br />
Stéphanie DEVAUD<br />
Walter MATTHEY-DORET<br />
Fabien ROBERT<br />
Santo ALI<br />
Christian LINDINGER<br />
PTR-MS Igls / IBk January 2003 23