Dietary exposure to heterocyclic amines in high-temperature cooked meat and fish in Malaysia

Food Additives and Conta m inan ts

Vol. 27 , No.8 , August 201 0, 1060- 1071

Q

ｾ＠ Taylor &Francis Taylor£..Fra nclsGroup

Dietary exposure to heterocyclic amines in high-temperature cpoked meat and fish in Malaysia

1 \. ." '" ｣Ｌ ｾ＠

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M.H.A. Jahurul, S. Jinap*, SJ. Ang , A. Abdul-Hamid, P . Hajeb. H.N. Lioet _{and I.S.M. Zaidul}

Cenl er of" Excellence /i)r Food S a/ i' ly Research (C EFS R ) . Fa cultv

0/

Foo d Scien ce and Technology, Universili Putra Malaysia. 43400 U PM , S elango r, Malavsia

(R ecci l'cd ]5 janllan ' 20 11!. /il1a l I'crsiun received 2] M arch 20 10 )

The inta ke of hcterocycl ic amin es is influenced by th e am ount and type of mca t and fish inges ted, frequen cy of consumpti o n, cooking method s. cookin g tem pe rature, and du rati on of cooking. In thi s study, the dietary intake o f heterocycli c aminc s in Ma lays ia and th eir main sources were inves ti gated . Fo rt y-two samples of meat and fi sh were anal ysed by hi gh-pe rfo rm a nce liquid chromatograp hy with ph o todi ode array detector to determin e the concent rati o n of th e six predo min a nt heterocycli c amin cs, namely: 2-amino-3-meth ylimidazo[4 ,5-f]q uin ol in e (I Q ). 2-a mi no-3 A-dimet hylimi dazo[4. 5-f lq uino Ii nee 1\1 eI Q). 2-amino- 3,S-dimethylimidazo[4 , 5-flq uin ox alin c (M e l Qx) . 2-amin o-Ｓ ａ ＬｾＭ ｴｲｩｭ ･ｴ ｨ ｹ ｬｩｭｩ､ ｡ｺｯ ｛ ＴＬＵＭ ｦ＠

Jq

uinoxalin e (4,8- DiMeIQx), 2-amino-3 ,7,S-trim eth yl imida w [4. 5-1' Jq uin ox alin e (7 .S- D i Me l Qx). ,1 nd 2·am ino - I -mcth yl-6-phcnylimida zo[4,5-b]pyridinc

(PhIP). Di eta ry intak e dat a we re obtain ed lI sing a food-freq uen cy qu es ti o nn a ire wh en interviewing 600 Ma la ys ia n res po ndent s. Th e level of tota l heterocyc lIc a mi ne s in food sam pl es studies ranged fr o m not detected

to 38.7 ng g- I. T he a ve ra ge dail y intak e level of hete ro cycl ic am ine was 553.7 ng pe r capita day- I. The intake of

PhIP was th e hig he st. followed by MeI Qx a nd Ivl eIQ. T he result s revea l tha t fri ed a nd grill ed chicken were the

maj o r di eta ry so urce o f hetewcycl ic am in es ill ivlalaysia . However. the he terocycl ic a mine intake by the' Mala ys ia n po pu lati o n was lo \\'c r tha n th ose repor ted fr olll other reg io ns

Keywords: expos llt'c a,; ses slll cnt: he te rocycli c am in es: co o ked fo od,;

Introduction

Het e rocycli c a mines (HCA s) a re po ss ible human a nd animal carcin oge n s fo nned in cooked m eat an d fi s h during hi g h-te mp e ra ture c ookin g processes (Int e rnation a l A ge ncy for R esea rc h o n Cance r (IARC) 1993). Th e rAR C a lso reported that HCA s such a s Ｒ Ｍ ｾ［ ｭｩｮｯ ＭＳＬ ＴＭ､ｩｬＱｬ･ｴｨ ｹ ｬｩｬｬｬｩ､｡ ｺｯ ｬＴ Ｎ ＵＭ fJquinoline,. (MelQ) , 2-amino-3,8-dime th ylimd azo [4,5-fJquinoxalin e (MeIQx), and 2- a mino-l-meth yl-6-phe-n ylimidazo [4, 5-b) pyridiyl-6-phe-ne (PhIP) are po ss ible huma yl-6-phe-n carcinogens; and 2-am ino-3-me th ylimida zo[4,5-f]-quinoline (IQ) a s a probabl e ca rcinogen. H C A s a lso ca use cancer at a varie ty of multiple sites in lab o ratory a nimal species (EIBa y oum y et al. 1995), Se veral ca se -control studies in various p o pul a tion s h ave repo rted po sitive associati o n s between a highe r intak e of well-coo ked meat and th e risk of co lon cancer (Sinh a e t al. 1999), breast ca nce r (Sinha e t a l. 2000), lun g ca nce r (Sinha 1998) , and gas tric cance r (d e Stefani et a l. 199 8). Hum a n s arc ex p o sed to p o te nti a ll y to x ic s u bstances including natura lly occ urrin g muta geni c and carci n o -ge nic c o mpoun ds . On e's daily di e t lll ay c o nt ai n H C A s a nd polycyclic a romatic hydroca rbon s ( PAH s)

as a

res ult of high-tempe rature processi ng o f m eat a nd fi s h .

Bo t h gro ups a re eas il y found in grill ed and fri ed m eat, p o ultry an d fi s h (Gomaa et al. 199 3; Kaze rouni e t al. 200 I; J iigersta d a nd Skog 20(5), T he present study is foc llsed on H C A s. Fri edman (1996) reported that some H CA s pl ay a n im p o rtant rol e in th e taste, fla vour, a nd eve n s mell o f h ea ted meat. H C A s are form ed by th e pyro lys is of pr o te in a nd amino acids in pro tein-rich food s. Th e ir form a tion depend s on several factor s such a s tempe rature, c ooking method and time, the degree of d o ne ness, th e c oncentration of HCA prec ursors, and th e presence of co mpounds with enhancing or inhib-it o ry e ffects (Felt o n et al. 2000; Busquets et a L 2004) . Th e re has bee n a large variability of estimated intake of di e ta ry H C A s reported from different countries (s uc h as Japan , Singapore , Sweden , Switzerland , Spa in , th e Unit e d Kingdom, a nd the United States). Th e rea so n ma y be due to diffe rent cooking method s, cook in g te mpe rature. duration of cooking, different ques t io nnaire m a ke-up, and th e studied populations . ｈ ｯ｜｜Ｇ ･ ｜｣ｾ ｲＬ＠ fo r th e in ves tigation o f hum a n ca rcino gen s, an a cc ur a te assess m e nt of th e indi vidu a l co ns umption of H eAs is esse nt ia l. U p-t o-dat e . exte nsive research on H CA s has been p e rfo rm ed on th e W es te rn die t (Wakabay a shi et a l. 1992; S kog

199 3;

Kni ze e t al. 1995: Z il11ll1c rli et a l. 2001: Sk og a nd Sol ya kov 2002 ;

Ｊｃ ｯ ｲｲ ･ｳｾ

ｮ､ｩｮｧ＠

a uthor. Emails:jina p @ food.upm.edu .my:sjinap@gmail. co m

' Present address : Depa rtm ent of Foo d Science an d T ec hn ology, Bogor Agri c ultura l U niversity, Bogo r, Indo nes ia

ISSN 0265-20 3X pri nt/lSS N 1464- 51 22 o nl ine

ro 10 10 Ta yl or & Francis

DO l: tO I080i]944004 1003801 190

Cantwell et ai. 2004; Llobet et a!. 2006: Domingo et a!.

2007; Marti-Cid et ai. 2007). Fried meat is an

impor-tant source of exposure to HCAs in the Western diet.

Although HCAs are formed at ppb levels (ngg- I),

their intake may be dangerous for human health

because of the total amount of cooked meat or fish

consumed (Busquets et a1. 2004).

This is the first extensive study on HCA exposure in

Malaysia. The analysis of Malaysian foods for the

occurrence of HCAs

is

important because there is

worldwide human exposure to these compounds

according to many reports (Augustsson et a!. 1997:

Byrne et ai. 1998; Kobayashi et ai. 2002; Wong et ai.

2005). The aim of the present study was to determine

the level of major six HCAs - IQ, MeIQ, MelQx, 4,

8-DiMelQx, 7, 8-DiMeIQx and PhIP - in some of the

most commonly consumed foods in Malaysia in order

to estimate the exposure of the Malaysian population

to these toxic compounds. The assessment was

per-formed by combining the content of HCAs in cooked

food samples analysed in the laboratory with

informa-tion on food consumpinforma-tion from the food frequency

questionnaire.

Materials and methods

Study population and questionnaire

A cross-sectional survey was conducted in the central

zone of Peninsular Malaysia covering three locations:

Kuala Lumpur, Putra Jaya, and Selangor state. The

respondents comprised three main races: Malay

(59.5%), Chinese (29.8%), . and Indian (10.2%) , and

other ethl/ie (0.5%), which were almost similar in

proportion to the Malaysian population. A total

sample size of 600 adult respondents aged IS years

and above was selected randomly for the survey. One

adult was chosen to represent the pattern of the whole

household. The sample size was measured by using a

two-staged stratified random sampling method and the

following equation was applied in the sample size

estimation:

1.96

2

x P(I -

_P)

ｎ ＭＭＭＭｾＭＭ

-

de

(I)

where a Z-value of 1.96 for

p=0.05

(95% confidence

level);

P

is the estimated prevalence of the sample equal

to 0.5; and

d

is a margin of error of 0.1 for

±

I 0%.

In the first stage of stratified random sampling, 75

enumeration blocks were selected from living quarters

where each living quarter consists of between 80 and

120 houses. In the second stage of stratified random

sampling, eight households were selected from each

enumeration block. The enumeration blocks and living

quarters for the survey were provided by the

Department of Stati stics Malaysia .

Fo od Additives and Contaminants

1061

Study design and questiollnaire

The questionnaire was divided into three sections:

cooking utensils and cooking methods used at home;

demographic data of the respondent; and dietary

intake. Dietary intake was assessed using a food

frequency questionnaire with a 1-month recall. The

food frequency questionnaire included information on

food items, the frequency of food consumption and the

portion size of food ingested. Before the main survey,

a pilot survey with 60 respondents was performed in

the targeted areas to test question planning in the

qu es tionnaire.

Selection o/food items and samplillg offoods

Food item s were selected based on the literature

publi shed in peer-revievied journals, which included

all type s of piocessed (such as drying and smoking)

and cooked (such a s grilling, roasting and frying) foods

report ed to contain !-leAs . There were in total 42 types

of food item s; three samples were purchased from the

same stall at different times of the day. The selected

meat and fish dishe s for this study were fried fish, fried

chicken (deep-fried), chicken soup , chicken curry,

steamed fi sh. chi cke n cooked with chilli (masak

merah). chicken burger, {ish curry, fried chicken,

keropok kkor , chicken satay. chicken randang, fish

fried in chilli, nu gge t, fish in tamarind (asam pedas

Ikan), paprika chicken, sweet-and-sour fish, sausage,

fish soup , chicken in coconut milk gravy (ayam masak

lemak), fri ed salted fish, fish in coconut milk, chicken

percik

(ayalll

percik), fried beef, beef cooked with

chilli, roasted fish, beef curry, smoke fish, fish in soya

sa uce , beef burger, beef satay, beef rendang, chicken

golek (ayam golek), fish in tauClI sauce, beef soup,

chicken kurma, mutton satay, paprika beef, fish with

three navours , grilled chicken tandoori, mutton curry,

beef in coconut milk gravy (masak lemak daging),

barbecue-grilled chicken, black pepper-grilled chicken,

beef kurilla , and honey-grilled chicken.

Samples of all food items were purchased from

local food stalls in Kuala Lumpur, Selangor, and

Putrajaya. Upon arrival in the laboratory, samples'

bones were removed from the meat and fish , then

samples were homogenized using a Waring blender

(Dawsonville, GA , USA). The homogenized samples

were then pooled and mixed together before being

quarter sampled. The representative samples were

stored at -18

v

C until analysis.

Statistical analysis

1062 Af.H.A. Jahllrlll et aJ.

considered to be reliable if Cronbach's alpha was at least 0.7. The descriptive analysis, including the mean, median, mode, standard deviation, and frequency of food taken by respondents, was used to describe the basic features of the data collected from the survey. An independent sample {-test was used to compare the differences between two genders. Analysis of variance (ANOVA) was used to test the differences between different HCAs in different food items. A p

< 0.05 was

considered to be statistically significant.

Chemicals

Solvent and chemicals were of both high-performance liquid chromatography (HPLC) and analytical grade. The water was purified in an Elix-MilliQ system (Millipore, Bedford, MA, USA). All solutions were passed through a 0.45-l1m filter before injection into the HPLC system. HCAs standards, i.e . 2-amino-3-methylimidazo[4,5-f]quinoline (lQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MelQ), 2-amino-3 ,8-dimeth y I imidazo[ 4, 5-f]q uinoxaline (Mel Qx), 2-ami no- 3,4,8- trimethylimidazo[ 4,5-f]quinoxaline (4,8-DiMeIQx), 2-amino-I-methyl-6-phenylimidazo[4,5-b]pyridine (Ph I P), 2-amino- 3, 7 ,8-trimethylimi-dazo[4,5-f]qunoxaline (7,8-DiMeIQx), and 2-a111ino-3,4,7,8-tetramethylimidazo[4,5-f]quinoxaline (4,7,8-TriMelQx) as internal standard, we re purchased from Toronto Research Chemicals (Dowl1sview. ON, Canada). Empty Extrelut-20 extraction cartridges were provided by Merck (Darmstadt, Germany) and Oasis MCX cartridges (3

mli60

mg) by Waters (Milford , MA, USA). MCX cartridges were pre-conditioned with ethyl acetate (2 ml). The solute diatomaceo,us earth refill material was obtained from 1ST (Hengoed, UK).

Determination of LOD and LOQ

The limit of detection (LOD) and limit of quantifica-tion (LOQ) were obtained by performing seven deter-minations of the lowest acceptable standard concentration. LOD was calculated as three times the standard of deviation (SD); LOQ was ten times the SD of the average background signal.

RccIJl'er), test

RecO\ery assays were established by spiking known amounts of the matrices (200 and 500 ng g - I sample) of the HCA standard solution to samples. Seven replicates were conducted on each fortified sample. The recovery assays were calculated by comparing the concentration of the fortified samples with those of the unfortified samples.

RCA extraction

The extraction and clean-up of all six HCAs studied was carried out following the modified method of Gross and Griiter (1992) and Messner and Murkovic (2004). Samples

(I

g) spiked with internal standard (4,7,8-TriMcIQx, 250 ppb) were dissolved in 12ml of 1 M NaOH and the suspension was homogenized using magnetic stirring for 3 h with 500 rpm (Heidolph, MR 3001 K). The alkaline solution was mixed with diato-maceolls earth (13 g) , and then transferred into empty Extrelut columas. Ethyl acetate was used as the extraction solvent and elute was passed through the coupled Oasis MCX cartridges. The MCX cartridges were washed with 0.1 M HCI (2 ml) and MeOH (2 ml). The analytes were eluted with 2 ml of MeOH-concentrated ammonia

(19/1,

vjv). The sam-ples were evaporated to dryness under a stream of nitrogen and the final extracts were dissolved in 100 pi methanol just before analysis for HCAs. Recovery of HCAs was determined using the standards addition method. The standards mixture (200 and 500 ng g- I sample) was spiked to the chicken, beef, and fish samples before the extraction step. HCAs were sepa-rated using reversed-phase LC and a TSK gel ODS T M

column (25 em x 4.6 mm. 5 11m). The LC systems con-sisted of a Waters LC model 2690 (CA, USA) with a photo-diode array detector (Waters model 2996), equipped with a quaternary solvent delivery system under a Waters Controller model 600, an autosampl er (Waters model 717) , and an on-line vacuum degasser. The mobile phase consisted of solvent A, 0.0 I M triethylamine in water (adjusted to pH 3.3 with acetic acid), and solvent B, acetonitrile (HPLC grade; Merck). Analyses were carried out in a !!radient mode for 33 min. Gradient programming \;as per-formed as follows: at zero to 18 min it was 5% to 23% B in

A,

at 18- 25 min it was 23% B in

A,

and at 25--33 min it was 23% to 25% B in A. The flow rate of the mobile phase was I ml min - I.

Estimated dai{.v intake

The estimated daily intake levels of HCAs by an adult (ng day - I per capita) were calculated using the average of each meat and fish product, i.e, (Hajeb et a!. 2009):

Daily intake per person per day]

= mean concentration (ngg - I) of HCAs x amount of meat and fish products consumed per day (g per capita)

Results 'and discussion

Table I. Socio-demographi c chara c te ri stics of respondents.

Demographic profil e Frequency %

Gender

Male

303

50.5

Female

297

49.5

Elhniciry

Malay

357

62.5

Chinese

179

29.8

Indian

61

10.2

Othe rs

3

0.5

Age group (rears )

18- 24

130

22.2

25-29

9 1

15.2

30-34

64

10.7

35- 39

64

10.7

40--44

67

11.2

45--49

52

8.7

50- 54

48

8

55- 59

30

5

6064

23

3 "

.0

65- 69

20

3.3

ｾＷＰ＠ II 1.8

Educa r ion /('1'1'/

Uneducated

21

3.5

Primar y sc hoo l

74

12.3

Seco nd ary scho ol

280

46.7

College

105

17. 5

Bachelor deg ree

80

13.3

Postgrad uCJt e

14

2.3

Others

20

3.3

Carccr categories

Government se r va nt 56

9.3

Pri va te sec to r

185

30.8

Self-employed

78

13

Farm er I

0.2

Student

96

16

Hou sewife

129

21.5

Oth e rs

55

8.3

HOl/se/wId income .'

::,,1000

101

16.8

1000- 1999

131

21.8

20002999

140

23.3

3000- 3999

79

13 .2

4000--4999

61

10.2

:::5000

88

14.7

Frequency of" earillg OUI

Everyday

104

17.3

On ce a week

n

13

Two to three times a week 133

22.2

Two to three tim es CJ mont h

10 2

17

Once a month

136

22.7

Others

47

7.8

Cooking urensils

Stov e

592

98.7

Mi c rowa ve

265

44 .2

Oven

22 1

36.8

Grilled-stove

XO

13.3

Grill ed charcoal

64

10.7

Othe rs 23 3.8

( con lil1ued)

Food Additives and Contaminants

1063

Tabl e 1. Continued .

Demographic profil e Frequency %

Cooking merhod

Stir-fried

394

65.7

Dee p-fried

248

41.3

Boiling

204

34

Baking

200

33.3

Grilled

249

41.5

Roasted

255

42.5

Tabl e 2. R eliability summary of food frequency

que stionnaire.

Variabl e

Food consumption frequ e ncy Food consumption portio n size Buy o r coo k th e fo o d s Pla ces of fo od avail a bility

Cronbach's Alpha

0.730

0.634

0.902

0.840

The percent age for ma le respondents was 50.5% ; for

female res pondents it was 49.5%. There were 59.5%

Malay respondent s, 29.8% Chinese, 10.2% Indian,

a nd 0.5°;;, of other ethnic (Table I). Overall, the

cook ing method fa pparatus of respondents was mostly

by stove (98.7 % ), followed by microwave (44.2%) and

oven (36.8 % ). The hi gher usage frequency of stove,

mi crowave a nd o ven was probabl y due to their

con ve llience. A gri lied stove ( 13.3

%)

and grilled

charcoal (10.7%) were used less by respondents. On

the other hand , th e mos tly used type of cooking was

stir-fried (65.7 % ) followed by deep-fried (41.3%),

boiling (34 % ) and baking (33.3%). This fact showed

that respondents preferred to consume fried foods

ＨｳｴｩｾＧＭｦｲｩ･､＠

and deep-fried) rather than other types of

food (Table I) . HCAs are toxic compounds formed

during grilling and frying of meat products in oil

(Jagerstad et al. 1998). The gri lli ng/ barbecuing method

of food preparation has been shown to produce

significant level of HCAs (Keating and Bogen 2004).

In the United States, Sinha et al. (1995) reported the

highest level of PhlP in pan-fried or grilled/barbecued

chicken samples. In that study, grilled/ barbecued

chicken contained the highest levels of PhIP ranging

from 27 to 480 ngg - ' and pan-fried chicken contained

PhIP ranging from 12 to 70ngg-

1

In other studies in

[image:4.602.96.458.108.737.2]

r

1064 M.H.A. Jahuru/

et

al

(A)0.10-,--- -- - -- - - ,

0.08

0.06

:::l <{

0.04

Q

0. 02

1

)

0.00

x

a

x

wQ :;;:

'"

(5 ｾ＠ ' 0 <Xl ,

ｾ＠ ｣ｯ ｾ＠

I'-x

Q

(L

:r

(L

16.00 1800 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00

Minutes

( 8 ) 0 . 1 0 . . - - - ,

008

0.06

0.04

Q

'"

ｾ＠

x

'"

a

a.,

w

,..: (L

1

"

:r (L

0.02

000

16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00

Minutes

Figure I. Repre sentati ve HPLC · PDA chromatogram ora grilled chick e n salay samp le : (A) spiked and (8) unspiked. Results for

thi s sample were MelQx = 81 ngg··1 _{and PhlP = 30 .6 ng g -}I _{Peak s were detected at 265 nm.}

ReliabiHty was used to measure the internal

consis-tency of the study. It was measured based on the

numerical

coefficient

01'

reliability,

which

was

Cronbach's alpha (Table 2). The results given in Table

2 showed that the highest values of Cronbaeh's alpha

were 0.902 and 0.840 for methods and for places of food

availability, respectively. These were followed by 0.730

and 0.634 for food consumption frequency and portion

size, respectively. In this study , all values ofCronbach's

alpha were

ｾＰＮＶ＠

where they were all accepted in

exploratory factor analysis and also supported the

reasonable internal consistency of variables. Therefore,

the results yielded satisfactory intcrnal consistency

(coefficient alpha) reliabilities for the related variables.

The results of the food-fr equ ency questionnaire

revealed that the most popular food \\as fried fish

(72.83 %) and deep-fried chicken (68.67%) , followed

by chicken soup (57.67(Yo) and chicken curry (56.50%).

These findings showed that fish and chicken were the

food items of choice for most respondents and that

they may impose a risk of contaminants from

high-temperature cooked foods.

The sample clean-up by tandem SPE using Extrelut

and Oasis MCX was sLlccessful and yielded one

fraction of HCAs, which wa s subjected to further

determination by HPLC. Furthermore, the applied

HPLC method provided a good separation of HCAs

that make the quantification of each HCA clearly

possible. Figure I shows a representative

chromato-gram on the basis of identification and quantification

of individual HCA compounds. The identification of

the HCA compounds found in a sample is based on

their retention times and UV spectra. Figure 2 shows

the UV spectra of the six authentic HCAs in sample

spiked by FICA standards. All spectra observed in

sampl e analysis were in good concordance with the

spectra of the authentic compounds.

The detection limits of" HCAs were: IQ, 1.5 ngg-

l ; [image:5.597.148.471.77.436.2]

Food Additives and Contaminants

1065

IQ

001,1921 2 iOll Ｌｾ Ｌ＠

ｾ＠ ｾ＠

｜ ｾ

Ｌｾｾ｟Ｚ

ＧｾＲ ｾ｟ｬ Ｌ＠

__

ｾ＠

_ _ ...

3..;:.;:69),.;.:..-..4;.;;:J 8:.;;;;5'O....".j

-{)Oll,/i-...:... _ _ _ _ _ _ __ _ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - . l

Me lQ

0,02

19iUl 2 :2U

::J

ｾ＠ 0.0

-{l.O " -_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ....,.J

-O()15 ... ｟ｾ＠ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ｾ＠ _ _ ｾ＠ _ ___..J

4, 8-DiMe IQ x

329.1

]

7,8-DiMcIQx

PhlP

Figure 2. UV spectra of HCA standard s found in spiked sam ples.

recoveries of spiked samples demon strated the

accu-racy of the method s used in th e study.

Table 3 shows the six HCA s detected in the

analysed food sa mples. There was a significa nt

differ-ence (p

<

0 .05) in the amount of' the six HCA s between

the different food s studied. PhlP was the most

com-monly found HCA (24%1), followed by MeIQx (23%)

and MeIQ (12 %) in the st udi ed food items. Very lo w

level s of 4, 8-DiMclQ x and 7,8-DiMeIQx we re detected

in th e samples. The hi ghes t level of tota l HCAs wa s

found in grilled and fried dishes, i. e. grilled chicken

sa ta y (38.7 ngg - I).

followed

by

grill ed chick en

black pepper (24.3 ng g-I), g rilled chicken perci k

(24.1 ngg- I) and s tir-fried

chicken (23.4ngg- ' ).

The higher levels of HCA s found in grill ed chicken in

thi s st udy may be due to the cooking meth od s,

temperature and cooking tim e, a nd a lso the precursors

s uch as amino acid , sugar , crea tinin e and moisture

content of the chi cke n. The hi gh level of HCAs in

grilled a nd fri ed foods are pro babl y beca use these

fo od s were cooked uSin g high

temperature of

125- 25WC (Busqucts et a1. 2004) and for relatively

long periods of 20-40 min (Keating and Bogen 2004).

Therefor e, a more frequent usage of the stove for

fryin g m ay expose peop le to hi gher concentrations of

HCAs. In th e prese nt s tud y. the food sa mples for

a na lysis were purchased from different restaurant. At a

loca l food sta ll or resta urant th ere could be a

signif-ica nt va riation in cooki ng ti me. as wel l as th e

temper-ature of m ea t and fish products sol d betwee n one

restaura nt and another.

[image:6.595.120.518.82.488.2]

1066

M.H.A. lahurul

et a l.

Table 3. HCA s concen trations in meat and fi sh sa mpl es (ngg - I).

Food item

Chicken curry Chicken so up Chicke n in milk gravy C hicke n percik Ch icke n kurma Fried ch icken

Chicken cooked with c hilli Ch icke n ho ney

Chicken paprika Chicken bl ack pep pe r Chicken ba rbecue C hi c ke n tandoori Chicken burger C hicke n re ndang Chicken sa ta y Chicken go le k Sausage Nugget

Fi sh with three flavour Fi sh in milk gra vy Fi sh curry Fish so up Salted fi sh F ish in tamarind Fried fish in c hill .i Sweet so ur fi sh Fried fi s h

F ish in tauc u sa u ce Fish ill soya sa uce Steam fi sh Charcoal gr ill fish Keropok Iekor Bee f sa tay Beef kurma Bee f paprika Bee f c urr y Bee f burge r Bee f re ndan g Beef so up

Bee f in coco nut milk Mutt on salay Mutton curry

Noles: n .d .: Not detecte d.

Cook in g method IQ

Boiled n.d.

Boile d !l .d.

Boiled n.d.

Grilled n .d .

Fried cooked n.d .

Stir fried n.d.

Fried /cooked n .d.

Grilled !l .d .

Fried/cooked n .d.

G rilled n.d.

Grilled n.d.

Grilled n.d .

Fried Il.d .

Boiled Il.d.

Grilled n .d .

Boil ed l1.d .

Fried Il.d.

Fri ed n.d.

Boiled n .d .

Bo il ed n .d.

Bo iled Il .d .

Boiled n .d.

Fri ed n.d.

Fried /cooked n .d .

Fried /coo ked n.d .

Fried /coo ked n .d.

Deep fri ed n .d.

F ri ed/coo ked n .d.

F ri ed /cooked n .d.

Steam n .d.

Grilled n .d .

Fried n.d.

Grilled n.d .

Bo il ed n.d.

Bo il ed n .d.

Boiled n .d .

Fri ed n .ci.

Fri ed/cooked n .d.

Boikd n .d.

Boiled n.d .

Grilled n.d .

Boil ed n .d.

MeJQ n.d . n.d . n.d. n.d . n.d. n.d. n.d. n.d. n.d. n. d. n.d. n.d . n.d. n.d . n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d.

2.4

n.d. n.d. n.d.

1.4

3.8 3.3 n.d . 1.6 n.d. n.d. n.d. n.d. n.d. n.d . n.d. n.d. n.d. n.d. n.d.

M e lQ x

n.d . n.d . n.d. 14 .5 n.d .

23.4

n.d.

n .d .

n.d .

24.3 4.2

n.d.

n. d. n .d . 8. 1 n.d. S.5 n .d. n.d. n.d. n .d. n .d. n.d. n.d. n.d . n.d. 8.8 n.d. n.d . n.d. 1.7 n.d. n.d. n.d. n.d. n.d. n.d. 12.3 n.d. n.d. 1.5 n.d.

4 ,8 -DiM e IQx

n.d. n .d. !l.d. n.d. n .d. n.d . n.d. n.d. n.d. n .d. n. d. n.d. n.d. n.d . n.d . n.d. n. d. n.d. n.d. n .d. !l.d . n. d. n.d . n.d . n.d . n.d. n.d . !I .d. !I.d . n.d. 0.5 n.d . 0 .7 n .d. n.d . !I .d. !l .d. 0.7 n .d. !l .d. n.d . n.d. 7,8 -DiMeIQx

!I .d . n.d . !l.d. n.d. n.d. n.d. n.d . !I .d. n.d. n.d. 1. 6 n .d. n.d . n.d. !I .d. n.d. n.d. 1.5 !I .d. !l .d. !I .d . n .d. n .d .

n.d. n.d . n.d . n .d. 1. 6 n.d. n.d. 0.9 n.d. n.d . n.d. !l .d. n .d. 1.0 n .d. n.d .

n .d . !l .d. n .d. P hlP n.d . !I.d. n .d. 9.6 n.d. n.d . n.d .

n .d . n .d. n .d. 9.9

4.4

n.d. n.d. 30.6 n.d. n .d 11 . 1

n.d. n.d. n.d. n.d. 4.8 n.d. n .d. n.d. n.d . n .d. n.d . n .d. ｾ ａ＠ n .d. 13.3 n .d .

n.d. n.d. 15. 1 n.d. n.d . n.d. 14. 2 !I .d. Total n.d . !I.d . n.d. 24.1 n .d. 23.4 n.d. n.d. !l .d. 24.3 15.7

4.4

n .d. !l .d. 38.7 n.d. S.5 12.6 n .d. n .d. Il .d. n.d. 7.2 n .d. n.d. Il .d. 10.2

SA

3.3 n .d. 13.1 n.d .

14.0

n.d. n .d. Il.d .

16. 1

13.0 n.d. n.d. 15.7 n.d.

Valu es belo w the limit of detection (LOD) a re calcula ted as Ongg-1 _{LODs of: IQ = 1.5ngg- ' ; M eIQ = 0.5ngg-l ;}

M e lQx =0 .3 ngg-l ; 4 ,8 -DiMeJQx = 0.5 n gg- I; 7,8-Ditvle IQx = 0.9 ng g-l ; and PhlP = 1.5 ng g- I.

middle-aged (18- 34) or olde r (more th a n 55) genera ll y consume more meat a nd fi sh compa red with other groups of res pondents. On average, Malaysians con-sum ed 104.16 g of meat and fi sh di shes per da y. W o ng et al. (2005) reported th e ave rage d a il y intake of 108 .7 g da yl for a ll meat t y pes in Sin ga pore. Oth er popula tionba se d surveys ha ve re ported the dail y cons u mp -tion of meat from 80 to 160 g da y-l in the Un ited Sta tes , Swed en, New Zea la nd , and J a p a n (Allgllstsson et al. 1997; By rn e e ( al. 1998 ; K o ba yas hi e t a l. 2002; Ferguson 2002). Data fr o m th e present stu dy sh ow tha t food s contributing t o th e in take of

HCA s in Malaysia are simi la r to those found in Western countries.

[image:7.600.85.540.111.537.2]

r

Food Additives and Contaminants

1067

Table 4. Mcan daily intake of HCAs from meat and fish (ng per capita day-I) in Selangor, Malaysia.

Food item Total HCAs (ngg - I) Food consumption (g day-I)

Chicken curry Chicken soup

Chicken in coconut milk gravy Chicken percik

Chicken k urma Fried chicken

Chicken cooked with chilli Chicken honey

Chicken paprika Chicken black pepper Chicken ba rbecue Chicken tandoori Chicken burger Chicken rendang Chicken salay Chicken golek Sausage

Nugget

Fish with three flavour Fish in coconut milk Fish curry

Fish soup Salted fish Fish in tamarind Fri ed fish in chilli Sweet sour fi s h Fried fish

Fish in tauCll sauce Fish in so ya sauce St ealll fi s h Charcoal grill fish Keropok lekor Beef satay Beef kurma Beef paprika Beef curry Beef burger Beef rendang Beef soup

Beef in coconut milk gravy Mutton satay

Mutton curry

o

o

o

24.1

o

23.4

o

o

o

24.3 15 .7 4A

o

o

38.7

o

5.5 12.6

o

o

o

o

7.2

o

o

o

10.2 5.4 3.3

o

13 . 1

o

14 .0

o

o

o

16.1 13.0

o

o

15.7

o

2.34 2.56 2.80 0.08 0.40 8.0 3.10 0.4 2.34 0.08 0.20 0.08 3.38 2A8 2.16 0.18 6.96 8.10 1.60 1.0 0.38 0.8

136

13.28 10.42 1.40 6. 14 1.0 4.22 1.10 0.38 1.96 0.76 2.0 0.6 1.50 0.70 0.80 1.72 4.0 OA 1.0

fable 5. Review of previous studies on HCAs level in food (ngg-I) from different countries.

Range of reported

MeIQx concentration Range of reported

PhIP

(ngg-I) concentration (ng g-I) Country

rypes of meat

3eef 0- 1.5 0 -6 Switzerland

073 032 USA

0.2-1 .3 0 .8-2.0 USA

0.72.9 0.64. 8 Spain

0- 7.1 0 - 30 USA

01.9 019 USA

: hicken 0- 1.8 0- 327.6 USA

00.3 2.346.9 Spain

0-9 6- 41\0 USA

0 0 -2 .5 Switzerland

0--0 .6 0.0210 Sweden

'i s h < 1- 5 0- 73 Switzerland

0--0 .5 0-4.9 Switzerland

0- 0.9 < 001-22 Swed e n

Exposure (ng per capita day-I)

o

o

o

1.9

o

187.2

o

o

o

1.9 3.1 0.3

o

o

83.6

o

38.3 102.1

o

o

o

o

9.8

o

o

o

62.6 5.4 13.9

o

5.00

o

10.6

o

o

o

11.3 lOA

o

Reference

o

6.3

o

Zimmcrli ct al. (2001) Knize et al. (1994) Keating and Bogen (2004) Busquets et al. (2004) Sinha et al. (199R) Felton et al. (2004) Keating and Bogen (2004) Bu squets et al. (2004) Sinha et al. (1995) Zimmerli ct al. (2001)

Skog et al. (1997)

[image:8.602.75.547.118.599.2]

Table 6 . Estim a ted mea n da il y in take of so me H CAs in var io ll s countries

Sum of

Geogra ph ic p h[p MelQx DiMelQx three H CAs

location (ng pC I' capi ta day-I) (ng per capi ta day-I) (ng per.capita day - I) (ng pC I' capita day- I)

New Zealand

Swe d en 72 72 [ 6

S\vedcn Sweden

Switzerland 12+- 1 S6 72 --·85 13-73 ＲｕＹﾷｾ＠ 19

USA 7iU-I09.7 19.6- 32.7 lS-2.2 993-144()

USA 427

80.5

14.7 522.2

USA 158 .3-137 .5 52 .1 0-640 35-4 1 2139 - 2056

USA 285.5-457 .9 D-44.8 3.5-4.0 322-50G.7

USA

USA i60-2 1S lJ3-US 6.5- 10.7 ＲＵＹＮＵｾｊ ＶｊＮＷ＠

USA 539- 1029 l) 1 - 126 18.9-469 (,48.9-120 I C)

J apan WA7 98-1 1. 2 07-03 495--64.S

Japan S .. .100 30()-3900

Germany 63 34 2 10 .1

Singapore

Spain 344 29 13.6-I S. 1 3SS

Notes: "I nta ke '01' [Q, Me IQ , M elQx , DiMelQx, PhIP , AaC, MeAac:. T rp- P- l and Trp-P-2.

bl n lakc of rQ, MclQ, McIQx, D iM e lQx an d Ph iI' . cI Qx, MelQx a nd p h[p.

dS um of I Q, 7,8-DiMelQx, MeJQ , 4,8- D ilvleI Qx, MelQx, and PhIP . "Mean values for co nt ro l su bjec ts anc! colon ca nce r cases.

[Total H CAs include [Q, Mc[ Qx, D iMclQx, P hIl' and Aac:. gBasec! o n a 70 kg bod y we igh t.

hSUt11 o f P hIl' , MelQx, D iMclQx, AaC an d IQ. 'Total HCAs include Trp-P-l a nd MeIQ.

IMean intake of IQ, Mc IQ , Me IQx, 4,S- D iMe IQx, 7,8 -D iMe IQx, phI P and I FP.

ng kg- I

To tal HCAs body mass

(ng pCI' cap ita day-I) day- I R eference

976" T homso n et al. (1996)

16010 _{A ugu stsso n et a l. ( 1997)}

77 A ugustsso n ct a!. ( 1999)

N.5Jc _{O lsson e( al. (2005)}

20l)-·397d _{Zimme r! i et a!. (200 I )}

Sinha e( al. (200 I

r

9.i

Boge n anc! Keatin g (200 1)£

Si nha et a!. (2 000)" Byrne et a !. (1998)

IS20

26

Layton e( a l. ( 1995)£

Nowe ll et a !. (2002)e

I I. O .. 19.9b _{Keating a nd Bogen (2004)£}

1.06-1. 1' Kobayashi el a!. (2002)£

Wakabayash i el aL (1997) Rohrma n n a nd Becke r (2002)

49.95J Wong et al. (2005)

934k Busquets et al. (2004)

kS1ll11 of DMIP, Glll-P-2 , IQ, Mc1Qx. McIQ, Gl u-P - [ , 7,8 -Di M eIQx , 4,8 -D iMcIQx. Norh armJn, Ha rman, Trp-P- 2, Trp-P-l , P hIP , AaC, McAac:.

o

ｾ＠

ClC

'>

:-"

::r:;

:J:..

<..., ;::,

Ef

...,

ｾ＠

ｾ＠

[image:9.842.103.773.141.352.2]

from me a t and fL sh were lowe r th a n th ose reported in the United States in 1995. where the value was 1820 ng pe r ca pita day- I (Layton e t al. 1995). H owever, o u; estimates of dail y intake a re in ag reem e nt with the studies of Keating and Boge n (2004) ( 1393 n g per capita da y- I) in the United States, and Busquets et al. (2004) (934ng pe r capita day-I ) in Spain. H oweve r. a lower average intake of HCAs wa s al so reported in differe nt studie s from Swede n, Switzerland, and Jap a n , wh e re 160 ng per capita day -I (Au gus tss o n et a l. 199 7 ), 397 ng per capita day - I (Zimmerli et al. 2001) , and 77 ng per cap it il day - I (Kobayashi et al. 20(2) we re est imated . respecti vel y. Table 6 s how s th e est imat ed mea n dail y intakes of so me HCAs in va ri o u s co un t ri es. Due to difference s in stud y d es ign, intake frequency . cook in g method , and th e duration o f cook in g . it is difficult to mak e a n accura te co mpari son o f th e dai ly int a kes of HCAs between thi s st ud y and other repo rt ed data from other studies. N eve rth eless, in viell of the more fried and grilled meat products and th e va ri ety of cooking methods for meat and fish, th e estim a ted inta kes of H C As in the prese nt study were almo st similar to tho se reported by other studies (Table 6).

This eva luation of HCA exposure showed lh al fried a nd grill ed mea t see m to be the largest so urces of HCA s in the di et in Selangor, and chicken wa s the grea tes t so urce of HCAs a mon g diffe rent typ es of meal. From our sur vey, the ratio of tota l chi cken intake per da y was found to be hi g he r tha n all the o the r meat inta kes. Moreove r, the concentration s or HCA s in chicken we re relatively high e r than any ot her meal. Th e m os t importai1t contribution o f these tindin gs is the identification of th e m aj o r contributors of HCAs in th e Malaysian diet. In this study , amo ng the 42 food ite ms analysed, only ten samples accounted for more (han 90% of the total dietary ex pos ure to HCA s in the Mala ys ia n population. It is also noted that minor changes of food choice can ha ve a large impact on exposure to HCAs. There was n o t mu c h differc nce in the intake of HCAs observed between men and women; the reason is presumabl y because the prefe rence for meat was almost similar be tween them.

Conclusion

This study has concluded that barbecuing and frying produce relat ive ly high levels of HCAs in protein-ri c h food products in Malaysia. The key findin g was tha t fri ed and grill ed c hi cke n a re m a lo r dietary so urces of the total int ake of H CAs am o ng Mala ys ian s. On the other hand, th e cons umption of bo iled or fried cooked foods co ntri b utes ve ry little to HCA intake. Further stu dies 011 the estimated di e tar y int a ke of HCAs should be ca rri ed out to clarify th e rela tion ship be tween th e intake of HCAs and the ri sk of cancer amon g Malaysians.

Food Additives and Contaminants 1069

Acknowledgement

The authors are g rateful to th e Na ti o n al Cancer C ou ncil (MAKNA) of Malaysia fo r its finan cia l support which allowed th em to carr y out the s tu dy .

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