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68%67,787(6,1)/8(1&(2)5$:0$7(5,$/
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+DQQD0RMVND ,ZRQD*LHOHFLVND
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$%675$&7
Background. Many animal studies have shown that acrylamide is both neurotoxic and carcinogenic. The first reports of
DFU\ODPLGHDFWXDOO\KDYLQJEHHQIRXQGLQIRRGVWXIIVZHUHSXEOLVKHGLQE\WKH6ZHGLVK1DWLRQDO)RRG$JHQF\LQFRQjunction with scientists from the University of Stockholm. It has since been demonstrated that acrylamide arises in foodstuffs
by the 0DLOODUGUHDFWLRQLHEHWZHHQIUHHDVSDUDJLQHDQGUHGXFLQJVXJDUVDWWHPSHUDWXUHV!&&RIIHHLQIDFWIRUPV
RQHRIWKHSULQFLSDOGLHWDU\VRXUFHVRIDFU\ODPLGHZKHUHLWLVQRUPDOO\GUXQNLQODUJHTXDQWLWLHVWKURXJKRXWPDQ\FRXQWULHV
ZRUOGZLGHWKDWLQFOXGHV3RODQG7KXVLWFRQVWLWXWHVDPDMRUGLHWDU\FRPSRQHQWLQDZLGHUDQJHRISRSXODWLRQJURXSVPDLQO\
ranging from late adolescents to the elderly.
Objectives. To determine the acrylamide level in commercial samples of roasted and instant coffee and in coffee substitutes
by LC-MS/MS method. The influence of coffee species and colour intensity of coffee on acrylamide level was also detailed.
Materials and methods. $WRWDORIVDPSOHVRIFRIIHHZHUHDQDO\VHGZKLFKLQFOXGHGWKDWZHUHJURXQGURDVWHGFRIIHH
11 instant coffees and 3 coffee substitutes (grain coffee). Analytical separation of acrylamide from coffee was performed
by liquid chromatography followed by tandem mass spectrometry (LC-MS/MS). To evaluate the colour intensity of ground
roasted coffee and instant coffee we used method of arranging (sequence).
Results. The highest mean acrylamide concentrations were found in coffee substitutes (818 g/kg) followed by instant coffee
JNJDQGWKHQURDVWHGFRIIHHJNJ2QHVLQJOHFXSRIFRIIHHPOGHOLYHUHGRQDYHUDJHIURPJDFU\ODPLGHLQURDVWHGFRIIHHWRJLQFRIIHHVXEVWLWXWHV7KHUHZHUHQRVLJQLILFDQWGLIIHUHQFHVLQDFU\ODPLGHOHYHOEHWZHHQ
the coffee species ie. $UDELFD vs 5REXVWD or a mixture thereof. The various methods of coffee manufacture also showed no
differences in acrylamide (ie. freeze-dried coffee vs agglomerated coffee). A significant negative correlation was observed
EHWZHHQDFU\ODPLGHOHYHOVDQGWKHLQWHQVLW\RIFRORXULQURDVWHGFRIIHHWKLVZDVQRWWKHFDVHKRZHYHUIRULQVWDQWFRIIHH
Conclusions. It was demonstrated that roasting process had the most significant effect on acrylamide levels in natural
FRIIHHKRZHYHUWKHUHZHUHQRUHODWLRQVKLSVIRXQGZLWKFRIIHHVSHFLHV'XHWRWKHKLJKDFU\ODPLGHOHYHOVGHPRQVWUDWHGLQ
FRIIHHVXEVWLWXWHVUHFRPPHQGHGDPRXQWVVKRXOGEHGHILQHGDQGPDQXIDFWXUHUVVKRXOGEHREOLJHGWRUHGXFHVXFKOHYHOVLQ
these products.
Key wordsDFU\ODPLGHGLIIHUHQWW\SHVRIFRIIHH/&0606PHWKRGFRIIHHVSHFLHVFRORULQWHQVLW\

675(6=&=(1,(
:SURZDG]HQLH:OLF]Q\FKEDGDQLDFKQD]ZLHU]WDFKZ\ND]DQRHDNU\ORDPLGPDG]LDDQLHQHXURWRNV\F]QHLNDQFHURJHQQH:NZLHWQLXU6]ZHG]ND1DURGRZD$JHQFMDGV\ZQRFLRUD]QDXNRZF\]8QLZHUV\WHWXZ6]WRNKROPLHSR
UD]SLHUZV]\RSXEOLNRZDOLGDQHR]DZDUWRFLDNU\ORDPLGXZSURGXNWDFKVSR\ZF]\FK$NU\ORDPLGSRZVWDMHZ\ZQRFL
w reakcji 0DLOODUGDSRPLG]\ZROQDVSDUDJLQLFXNUDPLUHGXNXMF\PLSRGZS\ZHPWHPSHUDWXU\!&:\VRNLH
VSR\FLHWHJRQDSRMXZZLHOXNUDMDFKW\PWDNHZ3ROVFHVSUDZLDHSURGXNWWHQPRHE\]QDF]F\PUyGHPDNU\ORDPLGXZGLHFLH]ZDV]F]DXRVyEGRURV\FKDOHWDNHVWDUV]HMPRG]LH\
&HOEDGD2]QDF]HQLH]DZDUWRFLDNU\ORDPLGXZUyQ\FKURG]DMDFKNDZ\PHWRG/&0606RUD]RFHQDZS\ZXJDWXQNX
NDZ\VSRVREXSURGXNFMLRUD]EDUZ\SURGXNWXILQDOQHJRQDSR]LRPEDGDQHJR]ZL]NX
0DWHULDLPHWRG\0DWHULDGREDGDVWDQRZL\SUyENLNDZ\ZW\PSUyEHNPLHORQHMNDZ\SDORQHMSUyEHNNDZ\
UR]SXV]F]DOQHMLSUyENLVXEVW\WXWyZNDZ\NDZD]ERRZD$NU\ORDPLGR]QDF]RQRPHWRGFKURPDWRJUDILLFLHF]RZHM
VSU]RQHM]WDQGHPRZVSHNWURPHWULPDV/&0606'RRFHQ\EDUZ\SUyEHNNDZ\Z\NRU]\VWDQRPHWRGNROHMQRFL
(szeregowania).

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3RZVLVND6WUHHW:DUVDZ3RODQGSKRQHID[
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:\QLNL :UyG SU]HEDGDQ\FK SUyEHN UyQ\FK URG]DMyZ NDZ\ QDMZ\V] UHGQL ]DZDUWR DNU\ORDPLGX VWZLHUG]RQR
ZNDZLH]ERRZHMJNJDQDVWSQLHZNDZLHUR]SXV]F]DOQHMJNJLNDZLHSDORQHMJNJ-HGQDILOLDQND
POQDSDUXNDZ\GRVWDUF]DDZ]DOHQRFLRGURG]DMXNDZ\SU]HFLWQLHRGJNDZDSDORQDGRJNDZD
]ERRZDDNU\ORDPLGX1LHVWZLHUG]RQRLVWRWQ\FKUyQLFZ]DZDUWRFLDNU\ORDPLGXZ]DOHQRFLRGJDWXQNX]LDUQD$UDELFD
vs 5REXVWD vs mieszanka $UDELFDi 5REXVWD). Sposb produkcji kawy rozpuszczalnej (liofilizowana vs. aglomeryzowana)
QLHZS\ZDQD]DZDUWRDNU\ORDPLGXZSURGXNFLHILQDOQ\P6WZLHUG]RQRLVWRWQSXMHPQNRUHODFMSRPLG]\
]DZDUWRFLDNU\ORDPLGXDLQWHQV\ZQRFLEDUZ\NDZ\SDORQHM=ZL]NXWDNLHJRQLHVWZLHUG]RQRGODNDZ\UR]SXV]F]DOQHM
:QLRVNL8]\VNDQHZ\QLNLSRWZLHUG]DMHSURFHVZ\SDODQLDNDZ\PDQDMLVWRWQLHMV]\ZS\ZQD]DZDUWRDNU\ORDPLGX
ZNDZLHQDWXUDOQHM1LHSRWZLHUG]RQRQDWRPLDVW]ZL]NXSRPLG]\SR]LRPHPDNU\ORDPLGXDJDWXQNLHPNDZ\=XZDJLQD
Z\VRN]DZDUWRDNU\ORDPLGXZVXEVW\WXWDFKNDZ\NDZD]ERRZD]DVDGQHZ\GDMHVLRSUDFRZDQLHZDUWRFLZVNDQLNRZ\FKGODWHMJUXS\SURGXNWyZVSR\ZF]\FKRUD]SRGMFLHSU]H]SURGXFHQWyZG]LDDQDU]HF]REQLHQLDDNU\ORDPLGX
szczeglnie w tym rodzaju kawy.
6RZDNOXF]RZHDNU\ORDPLGUyQHURG]DMHNDZ\PHWRGD/&0606JDWXQHNNDZ\LQWHQV\ZQREDUZ\

,1752'8&7,21
)RU RYHU WKH ODVW  \HDUV DFU\ODPLGH KDV EHHQ
manufactured on an industrial scale as a substrate for
V\QWKHVLVLQJSRO\DFU\ODPLGHSRO\PHUVWKLVEHLQJXVHG
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DVZHOODVLQWKHSDSHUWH[WLOHDQGFRVPHWLFVLQGXVWULHV
Numerous studies have demonstrated that acrylamide is
neurotoxic and may damage the central and peripheral
nervous systems (respectively CNS and PNS) in both
animal models and humans exposed to acrylamide at
WKHZRUNSODFH>@
Cell culture studies show that acrylamide is genotoxic and amongst other things it results in chromosomal
DEHUUDWLRQVVLVWHUFKURPDWLGH[FKDQJHJHQHSRO\SORLG\
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Animal studies demonstrate an increase in multi-organ
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gland when acrylamide was supplied in the drinking
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Research on Cancer had assigned acrylamide to those
substances that are probably carcinogenic to humans
JURXS$>@7KH-RLQW)$2:+2([SHUW&RPPLWWHHRQ)RRG$GGLWLYHVLQVWDWHGWKDWWKHHVWLPDWHG
exposure limits to dietary acrylamide are low for such a
substance that exhibits genotoxicity and cancerogenicity
DQGWKXVFRQVWLWXWHVDKXPDQKHDOWKKD]DUG>@
,Q IRRGVWXIIV DFU\ODPLGH IRUPV LQ WKH 0DLOODUG
reaction between free asparagine and reducing sugars.
The main dietary sources of acrylamide are potato
SURGXFWV)UHQFKIULHVDQGSRWDWRFULVSVFHUHDOVHJ
EUHDGEUHDNIDVWFHUHDOIODNHVDQGFDNHVWRJHWKHUZLWK
FRIIHH>@
When foodstuff acrylamide levels were monitored
LQ(XURSHGXULQJLWZDVIRXQGWKDWDQDYHUDJHRIJNJZHUHSUHVHQWLQURDVWHGFRIIHHZLWKPD[LPXPRIJNJRISURGXFW0XFKKLJKHUDPRXQWV
ZHUHPHDVXUHGLQLQVWDQWFRIIHHPHDQRIJNJ

DQG LQ FRIIHH VXEVWLWXWHV PHDQ RI  JNJ

In the latter case the acrylamide content reached a
PD[LPXPRIDERYHJNJLQLQGLYLGXDOVDPSOHV
>@8SWLOOQRZLQ3RODQGWKHUHLVKRZHYHUDODFNRI
any data on acrylamide levels in instant coffee and in
coffee substitutes.
Many studies demonstrate that coffee to be a signiILFDQWGLHWDU\VRXUFHRIDFU\ODPLGHHVSHFLDOO\LQWKRVH
countries with a high coffee consumption. The presence
RIFRIIHHLQDQDGXOWGLHWPD\GHOLYHUXSRIWKHHQWLUHGDLO\LQWDNHRIDFU\ODPLGH>@
:KHQ FRIIHH LV URDVWHG WKH ODUJHVW DPRXQWV RI
acrylamide form at the initial stage and swiftly attain
PD[LPXPOHYHOV1HYHUWKHOHVVWKHORQJHUWKHURDVWLQJ
FRQWLQXHVWKHQDFU\ODPLGHOHYHOVGHFUHDVH>
@,WDOVRDSSHDUVWKDWWKHDPRXQWVRIDFU\ODPLGHPD\
depend on the coffee species. When the acrylamide
content of $UDELFD and 5REXVWDFRIIHHVDUHFRPSDUHG
WKHQWKHODWWHUVKRZUDLVHGOHYHOVLQPRVWFDVHVKRZHYHU
WKHVHDUHQRWVWDWLVWLFDOO\VLJQLILFDQW>@$QRWKHU
SRLQWWKDWQHHGVVWUHVVLQJLVWKDWDWSUHVHQWWKHUHDUHDV
yet no effective tools available for decreasing acrylaPLGHOHYHOVLQFRIIHHXQOLNHWKRVHLQWKHRWKHUDIIHFWHG
IRRGVWXIIVVXFKDVFHUHDOVDQGSRWDWRSURGXFWV>@
The aim of this study was to determine the acrylamide level in commercial samples of roasted and instant
coffee and in coffee substitutes by LC-MS/MS method.
The influence of coffee species and colour intensity of
coffee powder on acrylamide level was also detailed.

0$7(5,$/$1'0(7+2'6
6DPSOLQJ
$QDO\VHVZHUHSHUIRUPHGRQFRIIHHVDPSOHVFRQVLVWLQJRIVDPSOHVRIJURXQGURDVWHGFRIIHHLQVWDQW
coffee and 3 coffee substitutes made by various manufacturers. The roasted coffee samples were randomly
selected as part of a monitoring programme in accordance with European Commission (EC) recommendations

Nr 3

Acrylamide in coffee and coffee substitutes

1R(&>@7KLVKDGEHHQSHUIRUPHGE\WKH
6WDWH6DQLWDU\,QVSHFWRUDWHGXULQJE\VDPpling from grocery shops and supermarkets throughout
WKHZKROHRI3RODQG7KHVDPSOHGFRIIHHZHUHIURP
brands that originated from 8 manufacturers of which
3 were Polish coffee roasting companies. One sample
comprised at least two commercially available packages
RIWKHVDPHQDPHPDQXIDFWXUHUDQGEDWFKQXPEHULQWKH
minimum quantity of 1 kg. The methods of sampling and
amounts taken were in accordance with EC Regulation
3DUW%1R>@
7KHURDVWHGFRIIHHVDPSOHVFRQVLVWHGRI$UDELFD
species (&RIIHD$UDELFD5REXVWD (&RIIHDFDQHSKRUD)
DQGZHUHDPL[WXUHRIERWK7KHUHZHUHVDPSOHVRI
instant coffee of which 9 were a mixture of $UDELFD and
5REXVWD. The coffee substitutes samples were principally
composed of roasted cereals (ie. rye and/or barley) and
chicory root. One sample contained also roasted beetroot.
In accordance with specifications provided by manufactuUHUVWKHGHJUHHRIFRIIHHURDVWLQJZDVPRGHUDWHLQWZHQW\
IRXUVDPSOHVOLJKWWRPRGHUDWHLQWZRFDVHVDQGOLJKWWR
GDUNDOVRLQWZR2IWKHHOHYHQLQVWDQWFRIIHHVDPSOHV
seven were freeze-drying whilst four were agglomerated.
All samples were transported and stored under conGLWLRQVVWDWHGRQWKHSURGXFWODEHOV3ULRUWRDQDO\VLV
sample replicates were mixed together from which a
representative portion was taken which then constituted
the laboratory sample.
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The laboratory samples consisted of 1.8 to 3 g
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the organic layer was removed. The aqueous layer was

incubated in an ultrasonic water bath. After cooling the

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were added. The sample was then further purified on
DFDUERQFROXPQ%DNHUERQG&DUERQPJPO
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so obtained was evaporated to dryness under nitrogen
and then reconstituted in 1 ml of mixture of water and
PHWKDQROYYRIZKLFK/ZDVXVHGDVWKH
injection volume for chromatographic analysis.
$QDO\VLVRIDFU\ODPLGHLQFRIIHHE\/&0606
Acrylamide in coffee was determined a previously
RSWLPLVHGDQGYDOLGDWHG/&0606PHWKRG>@&KURmatographic separation was achieved by the Dionex
8OWL0DWHV\VWHPRQD+\SHUFDUEFROXPQPP
[PPP7KHUPR6FLHQWLILFDIWHUDJXDUGFROXPQ
with the same absorbent. Analytical conditions: flow rate
RI  /PLQ FROXPQ WHPSHUDWXUH RI & VDPSOH
LQMHFWLRQYROXPH/DQGDQLVRFUDWLFPRELOHSKDVH
RIZDWHUPHWKDQROYYZLWKIRUPLFDFLG5XQ
time was 5 minutes. Detection and quantification were
SHUIRUPHGRQDQ$%6FLH[47UDSPDVVVSHFWURPHWHU
XVLQJPXOWLSOHUHDFWLRQPRQLWRULQJ050LQWKHSRVLWLve ion mode with an electrospray capillary voltage (IS) of
97KHFDUULHUJDVZDVQLWURJHQ&85 DQGWKH
LRQVRXUFHWHPSHUDWXUHZDV&$VSUHYLRXVO\GHVFULEHG>@DFU\ODPLGHFRQFHQWUDWLRQVZHUHFDOFXODWHG
by the peak areas under the curve originating from the
IROORZLQJLRQWUDQVLWLRQVP]$$P]
$$G3ZKHUHDVWKHLRQV$$
DQGP]$$G3) were used for verification.
$VVHVVLQJURDVWHGDQGLQVWDQWFRIIHHFRORXULQJ
This was performed according to an established
PHWKRGXVLQJDQPHWKRGRIDUUDQJLQJVHTXHQFH>@
+HUHDWHQPHPEHUWHDPUDWHGWKHJUDGHVRIFRORXULQJV
LQRUGHURILQWHQVLW\VKRZQLQSKRWRV 6DPSOHV

Photo 1. Samples of ground roasted coffee





175

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6DPSOHVVLJQLILFDQWO\PRGHUDWHO\GDUN
6DPSOHVVLJQLILFDQWO\GDUNHVW

176

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Nr 3

3KRWR 6DPSOHVRILQVWDQWFRIIHH




6DPSOHVVLJQLILFDQWO\OLJKWHVW
6DPSOHVVLJQLILFDQWO\PRGHUDWHO\GDUN
6DPSOHVVLJQLILFDQWO\GDUNHVW

were arranged by members in order of increasing intenVLW\DFFRUGLQJWRDWRVFDOHIRUURDVWHGFRIIHHDQG

a 1 to 11 one for instant coffee. Results were recorded
in custom-made reporting cards and then analysed
by .UDPHUV tables where the upper division served
to determine if there were any significantly lighter or
darker colourings in samples amongst all those tested.
The lower division was used to establish which of the
samples differed.
6WDWLVWLFDODQDO\VLV
Results were calculated on a Microsoft Excel spreDGVKHHWDQGVWDWLVWLFDOO\DQDO\VHGE\WKH6WDWLVWLFD
YHUVLRQ6WDWVRIW,QF$FU\ODPLGHFRQFHQWUDWLRQVDUH
given as the mean SD. ANOVA analysis was performed to first determine if any significant differences
existed between samples. When this proved to be the
FDVHDSRVWKRF1,5WHVWZDVXVHGWRILQGZKHUHWKH\
lay. Linear relationships were assessed by the 3HDUVRQ
correlation coefficient.

5(68/76
Concentrations of acrylamide in all the sample types
of commercial coffee studied are shown in Table 1.

The highest mean level values was in coffee substituWHVJNJIROORZHGE\LQVWDQWFRIIHHJNJ
whilst the lowest were for roasted coffee at 179 g/kg
(range: 61 397 g/kg). The ANOVA analyses indicated
that there were significant differences present between
VDPSOHVS
,QPRVWRIWKHURDVWHGFRIIHHVDPSOHVQ DFU\ODPLGHOHYHOVZHUHEHORZJNJRIZKLFKZHUH
ORZHUWKDQJNJ$FU\ODPLGHFRQFHQWUDWLRQVLQD
IXUWKHUVL[VDPSOHVUDQJHGEHWZHHQWRJNJ
whilst only two were higher at 386 and 397 g/kg (FiguUH)RUWKHLQVWDQWFRIIHHWKHUHZHUHVDPSOHVZLWK
DFU\ODPLGHOHYHOVEHORZJNJDQGRQHVWKDWUDQJHGEHWZHHQJNJ,QDIXUWKHUWZRVDPSOHV
DFU\ODPLGHOHYHOVZHUHDVKLJKDVDQGJNJ
)LJXUH$VDIRUHPHQWLRQHGFRIIHHVXEVWLWXWHVKDGWKH
KLJKHVWDFU\ODPLGHOHYHOVZKHUHWZRVDPSOHVIURPWKH
VDPHPDQXIDFWXUHUEXWIURPGLIIHUHQWEDWFKHVVKRZHG
DQGJNJZKHQFRPSDUHGWRDVDPSOHIURP
DQRWKHUPDQXIDFWXUHUDWJNJ
%HFDXVHDFU\ODPLGHLVKLJKO\ZDWHUVROXEOHLWFDQ
be assumed that it the entire acrylamide content of commercial products will be present in any coffee infusion.
Taking into account the variations of acrylamide levels
REVHUYHGDQVLQJOHFXSRIGULQNLQJFRIIHHLVH[SHFWHG

Table 1. Comparison of acrylamide level in different types of coffee and coffee substitutes
Type of coffee
ground roasted coffee1
instant coffee
coffee substitutes3

Acrylamide content
>JNJRIFRIIHH@
>JFXSRIFRIIHH@
x SD
min max
x SD
min max
179 85a*
61 397
a*

358 188b

b

c
c





VWDWLVWLFDOO\VLJQLILFDQWGLIIHUHQFHS
RQHWHDVSRRQJRIJURXQGURDVWHGFRIIHHZDVXVHGWRSUHSDUHDFXSRIFRIIHHPO

RQHWHDVSRRQJRILQVWDQWFRIIHHZDVXVHGWRSUHSDUHDFXSRIFRIIHHPO
3
RQHWHDVSRRQJRIFRIIHHVXEVWLWXWHVZDVXVHGWRSUHSDUHDFXSRIFRIIHHPO
*
1

177

Acrylamide in coffee and coffee substitutes

Nr 3
10
9

check value
for roasted coffee*

number of samples of roasted coffee

8
7
6
5
4
3
2
1
0
0

50

100

150

200

250

300

350

400

450

acrylamide content [g/kg]

)LJXUH 'LVWULEXWLRQRIVDPSOHVRIURDVWHGFRIIHHDFFRUGLQJWRWKHFRQWHQWRIDFU\ODPLGHQ 

*

VRXUFH>@
5

check value
for instant coffee*

number of samples of instant coffee

0
0

100

200

300

400

500

600

700

800

900

acrylamide content [g/kg]

)LJXUH 'LVWULEXWLRQRIVDPSOHVRILQVWDQWFRIIHHDFFRUGLQJWRWKHFRQWHQWRIDFU\ODPLGHQ 

*

VRXUFH>@
400
350

g/kg of product

300
250
188

200

180

162
150
100
50
0
Arabica (n = 6)

Robusta (n = 10)

mixture of Arabica and Robusta

(n = 12)

Figure 3. Comparison of acrylamide level in ground roasted coffee depending on the coffee species

178

+0RMVND,*LHOHFLVND

WRFRQWDLQEHWZHHQJLQURDVWHGFRIIHHWR
g in coffee substitutes (Table 1).
The mean amount of acrylamide present in the
$UDELFDURDVWHGFRIIHHZDVJNJZKLFKZDVORZHU
EXWQRWVLJQLILFDQWO\WKDQPHDQOHYHOVPHDVXUHGLQWKH
5REXVWDVSHFLHVDWJNJ)LJXUH
&RIIHHRULJLQDWLQJIURPURDVWLQJIDFLOLWLHVLQ3RODQG
KDGVOLJKWO\KLJKHUPHDQDFU\ODPLGHOHYHOVDWJNJ
FRPSDUHGWRWKRVHRIRWKHUPDQXIDFWXUHUVDWJNJ
but as before this difference was not significant.
/LNHZLVHWKHKLJKHUOHYHOVRIDFU\ODPLGHIRXQGLQ
freeze-dried coffee compared to agglomerated coffee
 JNJ YV  JNJ ZHUH DOVR QRW VLJQLILFDQW
7DEOH

Nr 3

7DEOH &RPSDULVRQRIDFU\ODPLGHOHYHOLQLQVWDQWFRIIHH
depending on the way of production
Acrylamide content
Number of
>JNJRIFRIIHH@
Type of coffee
samples
x SD
min max
freeze-dried
7


agglomerated




The intensity of coffee colouring in the studied

VDPSOHV DUH VKRZQ LQ SKRWRV  DQG  7KH UHVXOWV
demonstrate significant differences of intensity in
roasted coffees irrespective of the coffee species. In
IDFWDQHJDWLYHFRUUHODWLRQZDVREVHUYHGU 
SEHWZHHQDFU\ODPLGHOHYHOVDQGWKHLQWHQVLW\
RIFRIIHHFRORXULQJV)LJXUH7KHKLJKHVWVWDWLVWLFDOO\
VLJQLILFDQWSDFU\ODPLGHOHYHOVZHUHVHHQZKHQ
WKHFRORXUZDVOLJKWHVWDWJNJFRPSDUHGWRWKRVH

16

the intensity of the color of ground roasted coffee

14
r = -0.5569; p = 0.0386
12

10

0
100

150

200

250

300

350

400

450

acrylamide content [g/kg]

)LJXUH 7KHFRUUHODWLRQEHWZHHQDFU\ODPLGHFRQWHQWLQFRIIHHDQGFRORULQWHQVLW\RIJURXQGURDVWHGFRIIHH
12

the intensity of the color of instant coffee

10

r = 0.4616; p = 0.1530
2

0
100

200

300

400

500

600

700

800

900

acrylamide content [g/kg]

Figure 5. The correlation between acrylamide content in coffee and color intensity of instant coffee

179

Acrylamide in coffee and coffee substitutes

Nr 3

Table 3. Comparison of acrylamide level in roasted and instant coffee depending on the intensity of colour
Intensity of coffee color1
VLJQLFDQWO\OLJKWHVW
VLJQLFDQWO\PRGHUDWHO\GDUN
VLJQLFDQWO\GDUNHVW

5RDVWHGFRIIHHQ
Number of samples

5
5


x SD
*
168 55
197 69

,QVWDQWFRIIHHQ 
Number of samples
x SD


3
381 119



evaluated on the basis of .UDPHUVtables

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where the colour were moderately dark (197 g/kg) or

dark (168 g/kg) (Table 3).
,Q FRQWUDVW WR URDVWHG FRIIHH WKHUH ZDV QR UHODtionship observed between acrylamide content and the
intensity of colour for instant coffee (Figure 5). Less
acrylamide was in instant coffee with the lightest coloXULQJVJNJWKDQFRIIHHZLWKPRGHUDWHO\GDUN
or dark colourings at 381 g/kg and 399 g/kg respecWLYHO\DOWKRXJKWKHVHGLIIHUHQFHVZHUHQRWVLJQLILFDQW
(Table 3).

',6&866,21
The mean acrylamide content in the roasted coffee
samples was 179 g/kg ranging from 61 to 397 g/kg
depending on the product. Similar findings were observed in a study by $QGU]HMHZVNLHWDO>@ZKLFKWHVWHG
GLIIHUHQWFRIIHHW\SHVDYDLODEOHRQWKH86$PDUNHW
ZKHUHDFU\ODPLGHOHYHOVUDQJHGIURPWRJNJ
Higher amounts of acrylamide were found in various
coffee types in Denmark by *UDQE\ and )DJK >@
$FFRUGLQJWRWKLVVWXG\>@RQHOLWUHRIH[SUHVVRFRIfee contains on average 9.5 g of acrylamide (range 7
WRJZKLFKLQWHUPVRIDNJRIFRIIHHJLYHVDQ
DYHUDJH HTXLYDOHQW RI  J LW ZDV DVVXPHG WKDW
JRIURDVWHGFRIIHHZHUHUHTXLUHGWRPDNHXSOLWUH
of drinking coffee). A smaller amount was demonstraWHGE\DQRWKHUVWXG\FRQGXFWHGLQ3RUWXJDO6RDUHVet
DO>@ZKHUHH[SUHVVRFRIIHHFRQWDLQHGDFU\ODPLGH
from 57 to 181 g/kg coffee (the authors claiming that
DQH[SUHVVRFRIIHHLVPDGHXSRIJH[WUDFWZLWKPO
ZDWHU +RZHYHU PXFK ORZHU DFU\ODPLGH FRQFHQWUDtions were demonstrated by 6HQ\XYDand *RNPHQ>@
LQ7XUNH\ZLWKOHYHOVLQJURXQGURDVWHGFRIIHHUDQJLQJ
IURPWRJNJ
The mean levels of acrylamide in instant coffee were
found to be 358 g/kg which was twice that in roasted
coffee (179 g/kg). Similar results were obtained by
RWKHUVWXGLHV$QGU]HMHZVNLHWDO>@DQG6RDUHV et al
>@ ,Q WKH 86$ WKH DFU\ODPLGH FRQWHQW RI LQVWDQW
FRIIHHUDQJHGIURPWRJNJZKHUHDVWKHPHDQ
acrylamide levels in commercial products in Portugal
ZHUHJNJ>@/RZHUDFU\ODPLGHFRQFHQWUDWLRQV
than those presented were seen in instant coffee studies
IURP7XUNH\>@LHRIWRJNJ6RPHZKDWKL-

gher acrylamide levels (8 g per litre of drinking coffee)

ZHUHVHHQLQWKHDIRUHPHQWLRQHG'DQLVKVWXG\>@RI
*UDQG\ and )DJK7DNLQJLQWRDFFRXQWWKDWJRI
FRIIHHDUHXVHGSHUOLWUHLQGULQNLQJFRIIHHWKHQLQVWDQW
coffee in the Danish study contained 597 g acrylamide/kg of commercial product. A monitoring study
FRQGXFWHGWKURXJKRXW(XURSH>@GXULQJ
GHPRQVWUDWHGVOLJKWO\KLJKHUDFU\ODPLGHOHYHOVZKHUH
RQDYHUDJHLQVWDQWFRIIHHFRQWDLQHGJNJUDQJLQJ
EHWZHHQWRJNJ
The presented study showed that coffee substitutes
FRQWDLQ RQ DYHUDJH  WLPHV WKH DPRXQW RI DFU\ODmide found in roasted coffee. The results are similar
to that demonstrated in the aforementioned European
VWXG\>@ZKHUHWKHPHDQDFU\ODPLGHOHYHOVIRXQGLQ
FRIIHHVXEVWLWXWHVZHUHJNJZLWKDPD[LPXP
RI  JNJ$FU\ODPLGH FRQFHQWUDWLRQV LQ FRIIHH
substitutes depend on the composition of products.
These products contain chicory root and roasted ceUHDOVHJU\HEDUOH\DQGVRPHDOVRURDVWHGEHHWURRW
All these ingredients contain acrylamide precursors ie.
reducing sugars such as glucose and fructose as well as
asparagine. The differences in acrylamide levels found
may be accounted for by the variety of coffee substitutes sampled and of having different manufacturers
and differing raw materials that thereby influence the
ILQDO FRPSRVLWLRQ RI SURGXFW )RU H[DPSOH FKLFRU\
URRWVFRQWDLQWZLFHDVPXFKIUXFWRVHDVJOXFRVH>@
where the former is more reactive than the latter during
DFU\ODPLGHIRUPDWLRQ)XUWKHUPRUHLWKDVEHHQQRWHG
by :RURELHMand 5HOLG]\VND>@WKDWWKHUHDUHODUJH
differences in reducing sugars present in coffee substiWXWHVZKLFKWKXVHIIHFWWKHOHYHOVRIDFU\ODPLGHIRXQG
in a given final product.
It should be stated that none of the roasted coffee
or instant coffee samples showed acrylamide levels
WKDWH[FHHGHGWKHFKHFNYDOXHRIDQGJNJ
UHVSHFWLYHO\ DV VWDWHG LQ (& UHFRPPHQGDWLRQV IURP
>@$WSUHVHQWWKHUHDUHKRZHYHUQRVXFKFKHFN
values existing for coffee substitutes.
Although the study showed that acrylamide levels
were lower in $UDELFD coffee than in 5REXVWD or mixtuUHVWKHUHRIWKHVHGLIIHUHQFHVZHUHQRWVLJQLILFDQW+RZHYHUOLNHILQGLQJVKDYHEHHQVKRZQLQDQRWKHUVWXG\
by /DQW]HWDO>@ZLWKDFU\ODPLGHOHYHOVRIDQG
JNJLQUHVSHFWLYHO\5REXVWD and $UDELFDFRIIHH



+0RMVND,*LHOHFLVND

WKHGLIIHUHQFHVDOVREHLQJQRWVLJQLILFDQW,QDGGLWLRQ
a study by $OYHVHWDO>@GHPRQVWUDWHGWKDW5REXVWD
coffee had significantly higher acrylamide than $UDELFD
FRIIHHYVJNJWKHODWWHUGHSHQGLQJRQ
the country from which the beans originated. It appeDUVWKDWGXULQJURDVWLQJPRUHDFU\ODPLGHLVIRUPHGLQ
5REXVWD coffee than $UDELFDFRIIHHEXWXSWLOOQRZWKH
determining factors remain unexplained. Nevertheless
it has been suggested that fresh 5REXVWD coffee beans
FRQWDLQPRUHDVSDUDJLQH>@WKDQ$UDELFD ones.
The instant coffee products that appear on the Polish
PDUNHWYDU\LQWKHPHWKRGVRIPDQXIDFWXUHHVSHFLDOO\
in how the coffee is dehydrated. When coffee is freezeGU\LQJIURPIUR]HQH[WUDFWVWKHLFHLVUHPRYHGWKRXJK
ZDWHUHYDSRUDWLRQKRZHYHULQDJJORPHUDWHGFRIIHHWKH
extract is dried under streams of hot air up to the moment
that the dissolvable granules are formed. Despite such
GLIIHUHQFHVWKHFXUUHQWVWXG\GLGQRWVKRZDQ\VLJQLILcant association between these methods of production
ZLWKDFU\ODPLGHOHYHOVWKHDSSDUHQWLQFUHDVHRI
acrylamide in freeze-dried coffee was not significant.
$QHJDWLYHDQGVLJQLILFDQWFRUUHODWLRQSZDV
demonstrated between acrylamide concentrations and
the intensity of color which was consistent with other
ZRUN7DH\PDQVHWDO>@DQG*RNPHQ& 6HQ\XYD>@
Lower acrylamide levels in dark coloured coffee compared to the light coloured ones reflect the sensitivity of
DFU\ODPLGHWRGHFRPSRVHGXULQJURDVWLQJDFU\ODPLGH
is principally formed at the start of the roasting process
but its content decreases the longer the roasting contiQXHV>@(YHQWKRXJKWKHUHZHUHGLIIHUHQFHVLQ
FRORXULQJZLWKLQWKHLQVWDQWFRIIHHVDPSOHVWKLVZDV
found not to be linked to acrylamide concentrations.
The presented study confirms that the roasting
process is a key factor in determining the acrylamide
levels in the finished product. The species bean (ie. raw
PDWHULDO KDV D VPDOOHU HIIHFW DOWKRXJK LW ZRXOG EH
worth checking the acrylamide content in $UDELFD and
5REXVWD coffee according to country of origin.
It can be summarised that coffee forms a significant
source of dietary acrylamide within the population of
persons consuming high amounts of coffee. Of special
concern is the high acrylamide levels in coffee subVWLWXWHVZKLFKRIWHQWDNHVWKHSODFHRIQDWXUDOFRIIHH
especially in the more vulnerable population groups
such as children and pregnant or breast feeding women.
It is therefore vital that check values are established for
FRIIHHVXEVWLWXWHVWKDWQXWULWLRQDOUHFRPPHQGDWLRQVDUH
modified accordingly in affected groups and that targeWHGHGXFDWLRQLVXQGHUWDNHQ)XUWKHUPRUHEHFDXVHRIWKH
wide variations of acrylamide concentrations within the
VDPHJURXSRIFRIIHHSURGXFWVPDQXIDFWXUHUVVKRXOG
take whatever steps are necessary to reduce such levels.

Nr 3

&21&/86,216
1. The study findings confirm that coffee can be an
important dietary source of acrylamide. Amongst
WKHYDULRXVFRIIHHW\SHVFRIIHHVXEVWLWXWHVKDGWKH
highest mean acrylamide concentrations (818 g/kg)
followed by instant coffee (358 g/kg) and then by
roasted coffee (179 g/kg).
 :HGLGQRWILQGVLJQLILFDQWGLIIHUHQFHLQDFU\ODPLde content between coffee types or mixtures (ie.
$UDELFD vs 5REXVWD vs mixtures thereof) nor in the
methods of manufacturing instant coffee (ie. freeze-drying vs agglomeration).
3. We found a negative correlation between acrylamide
levels in roasted coffee and the intensity of color of
coffee. This was not observed with instant coffee.
 7KH SUHVHQWHG VWXG\ FRQILUPV WKDW WKH SURFHVV RI
roasting coffee beans is a significant factor in determining the acrylamide content in the final product.
,QFRIIHHVXEVWLWXWHVWKLVFULWLFDOIDFWRULVWKHFRPposition of the raw material.
Acknowledgements
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6WXGLHV WR GHWHUPLQH VXEVWDQFHV IRUPLQJ GXULQJ
IRRGVWXIISURFHVVLQJDQGWKHLULPSDFWRQKXPDQKHDOWK
XQGHUWDNHQE\WKH1DWLRQDO)RRGDQG1XWULWLRQ,QVWLWXWH
:DUVDZ3RODQG1R'.
Conflict of interest
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Assessment of acrylamide intake by duplikate diet study.
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8. &KXVWHF]NL3ZLGHUVNL).: Zastosowanie metod analizy
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 '\ELQJ(6DQQHU7: Forum. Risk Assessment of AcrylDPLGHLQ)RRGV7R[LFRORJLFDO6FLHQFHV
13. ()6$: Scientific Report. Results acrylamide levels in
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oncogenicity study in rats with acrylamide. Fundam.
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15. *LHOHFLVND,0RMVND+ Optimisation and validation
of the analytical procedure for the determination of
acrylamide in coffee by LC-MS/MS using SPE clean-up.
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17. *UDQE\.)DJW6.: Analysis of acrylamide in coffee and
dietary exposure to acrylamide from coffee. Analytica
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committee on food additive. WHO Technical Report

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3:-: Acrylamide exposure from foods of the Dutch
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