Dietary exposure to HMF from Norwegian food and correlations with urine metabolites of short-term exposure

T. Husøy, M. Haugen, M. Murkovic, D. Jöbstl, L.H. Stølen, T. Bjellaas,

C. Rønningborg, H. Glatt, J. Alexander

Graz University of Technology

Institute of BiochemistryFunctional Food Group

EUROPEAN

COMMISSION

FOOD QUALITY AND SAFETY

Food and Chemical Toxicology 46 (2008) 3697–3702

Metabolism of HMF

OOH O O

O OS

OO

O

OOH O

OH

Sulfotransferases

proposedpathway

Oxidation

mainpathwayin kidneys andurinary excretion

Retention time (min)

0 2 4 6 8 10 12 14 16

MS

re

spo

nse

(m

/z =

30

5)

Standard

Urine

Z/E-Isomers

ON

NH

HO

NO2

NO2

ON

HN

HO

NO2

NO2

DNPH-derivatives of HMF

Chromatogram of HMF-DNPH in urineRP, 400 nm

Retention time (min)

0 2 4 6 8 10UV

res

pons

e fo

r ur

ine

sam

ple

(mA

U)

-0,2

0,0

0,2

0,4

0,6

0,8

1,0

1,2

1,4

HM

F

standard (1 µg/ml)

urine (80 ng/ml)

Renal excretion of HMF

Time (h)

0 1 2 3 4 5 6 7

Exc

rete

d am

ount

(µ

g)

0

50

100

250

GC analysis of HMFA in urineafter silylation with BSTFA/TMCS

Retention time (min)8 9 10 11 12 13 14 15

Det

ecto

r re

spo

nse

(cou

nts)

0

1e+6

2e+6

3e+6

HMFA

standard

urine(m/z = 147)

urine (TIC)

Retention time (min)

0 1 2 3 4 5 6 7

Plasma

Coffee

280 nm255 nm

UV

ab

sorp

tion

HMF and HMFA

Chromatogram of HMF and HMFA in urine

Retention time (min)

0 1 2 3 4 5 6 7 8 9

Det

ecto

r re

spo

nse

(280

, 255

nm

)

HMF HMFA

255 nm280 nm

standard

urine

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 productsFood 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

coffee; 63

milk products; 11

juice; 9

bread; 7

dark beer; 4

others; 6

Contribution of single food groups to the uptake of HMF

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

Urinary excretion

HMFA 12.4 11.8 1.68 40.5

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)

rsp = Spearman’s rank correlation coefficient.

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 = 0.71 <0.001

Dried fruits rsp = 0.29 <0.05

Beer rsp = 0.33 <0.05

Husøy et al., 2008