Dietary exposure to HMF
from Norwegian food and correlations with
urine metabolites of short-term exposure
Graz University of Technology
Institute of Biochemistry
Functional Food Group
T. Husøy, M. Haugen, M. Murkovic, D. Jöbstl, L.H. Stølen, T. Bjellaas,
C. Rønningborg, H. Glatt, J. Alexander
EUROPEAN
COMMISSION
FOOD QUALITY AND SAFETY
Food and Chemical Toxicology 46 (2008) 3697–3702
Metabolism of HMF
proposed
pathway
HO
O
O
Sulfotransferases
O
O
S
O
O
O
O
Oxidation
main
pathway
in kidneys and
urinary excretion
HO
O
O
OH
DNPH-derivatives of HMF
NO2
HO
N
N
H
O
NO2
MS response (m/z = 305)
NO2
Z/E-Isomers
HN
HO
N
O
Urine
Standard
0
2
4
6
8
10
Retention time (min)
12
14
16
NO2
Chromatogram of HMF-DNPH in urine
HMF
UV response for urine sample (mAU)
RP, 400 nm
1,4
1,2
1,0
0,8
standard (1 µg/ml)
0,6
0,4
0,2
urine (80 ng/ml)
0,0
-0,2
0
2
4
6
Retention time (min)
8
10
Renal excretion of HMF
Excreted amount (µg)
250
100
50
0
0
1
2
3
4
Time (h)
5
6
7
Detector response (counts)
GC analysis of HMFA in urine
after silylation with BSTFA/TMCS
urine (TIC)
3e+6
2e+6
urine
(m/z = 147)
HMFA
1e+6
standard
0
8
9
10
11
12
13
Retention time (min)
14
15
HMF and HMFA
280 nm
UV absorption
255 nm
Plasma
Coffee
0
1
2
3
4
5
Retention time (min)
6
7
Chromatogram of HMF and HMFA
in urine
255 nm
Detector response (280, 255 nm)
280 nm
HMF
HMFA
urine
standard
0
1
2
3
4
5
6
Retention time (min)
7
8
9
Study design
• 47 non-smokers + 6 smokers (2 ♂, 4 ♀) (53 participants)
• 24 h dietary recall
• 24 h collection of urine
• 20 ♂ (45  13) + 33 ♀ (41  11)
• Foods (35) from the 24 h dietary recall were
purchased and analysed for HMF and HMFA
– 26 contained HMF and 10 HMF
Food groups containing HMF and HMFA
• Norwegian brown
cheese
• Jam and honey
• Cereal products
• Dried fruits
• Fruit juices
• Coffee
• Beer
• Coffee
• Cereal products
HMF and HMFA in Norwegian food products
Food product
HMF
(mg/kg)
HMFA
(mg/kg)
n
Norwegian brown cheese
0.08–0.21
nd
3
Soft brown cheese, with added sugar
0.20
nd
1
Honey
0.34
0.05
1
Cereals with dried fruits
0.37
nd
1
White bread
0.24
0.02
1
Bread
0.06 – 0.14
nd – 0.07
2
Crispbread, thin, made of rye
0.45
nd
1
Crispbread, wafer-thin
0.65
0.70
1
Wholemeal rusk
0.61
nd
1
Raisins
5.00
nd
1
100% apple juice
0.30
nd
1
Coffee, instant
91.3–3060
5.6–174
5
Cappuccino with added sugar, instant powder
1.72–143
0.23–17.2
3
Ground coffee
262–547
40.3–130
7
Beer
0.2–0.3
nd
2
Dark beer
13.3
nd
1
Prunes
237
nd
1
Canned peaches
5.8
nd
1
Husøy et al., 2008
Contribution of single food groups
to the uptake of HMF
milk
products; 11
juice; 9
bread; 7
dark beer; 4
others; 6
coffee; 63
Estimated 24 h dietary intake
and 24 h urinary excretion of HMFA
Dietary intake
(mg)
Mean Median
Min
Max
HMF
5.56
3.04
0.01
32.6
HMF + HMFA
6.32
3.58
0.01
37.0
HMFA
0.76
0.20
0.00
4.39
12.4
11.8
1.68
40.5
Urinary excretion
HMFA
Husøy et al., 2008
Correlation
between 24 h dietary intake of HMF, HMFA, HMF + HMFA, coffee, dried fruits and beer,
and 24 h urinary excretion of HMFA (n = 53)
24 h dietary intake
HMFA (mg) in 24 h urine
P value
HMF (mg)
rsp = 0.57
<0.001
HMFA (mg)
rsp = 0.60
<0.001
HMF + HMFA (mg)
rsp = 0.58
<0.001
Coffee (ml)
rsp = Spearman’s
rank correlation coefficient.
rsp = 0.71
<0.001
Dried fruits
rsp = 0.29
<0.05
Beer
rsp = 0.33
<0.05
Husøy et al., 2008