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Organochlorine pesticide andpolychlorinated biphenyl residues inhuman milk from Tabriz, IranR. Dahmardeh Behrooz a , A. Esmaili Sari b , N. Bahramifar c , F.Naghdi d & A.R. Shahriyari aa Faculty of Natural Resources, Department of EnvironmentalSciences, Zabol University, Zabol, Sistan, Baluchestan, Iranb Faculty of Natural Resources and Marine Sciences, Departmentof Environmental Sciences, Tarbiat Modares University, Nour,Mazandaran, Iranc Faculty of Sciences, Department of Chemistry, Payam NourUniversity, Sari, Mazandaran, Irand Faculty of Natural Resources, Department of EnvironmentalSciences, Science and Research University, Ahvaz, Khozestan, Iran

Available online: 23 Nov 2009

To cite this article: R. Dahmardeh Behrooz, A. Esmaili Sari, N. Bahramifar, F. Naghdi & A.R.Shahriyari (2009): Organochlorine pesticide and polychlorinated biphenyl residues in human milkfrom Tabriz, Iran, Toxicological & Environmental Chemistry, 91:8, 1455-1468

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Toxicological & Environmental ChemistryVol. 91, No. 8, December 2009, 1455–1468

Organochlorine pesticide and polychlorinated biphenyl residues in human

milk from Tabriz, Iran

R. Dahmardeh Behrooza*, A. Esmaili Sarib, N. Bahramifarc, F. Naghdid andA.R. Shahriyaria

aFaculty of Natural Resources, Department of Environmental Sciences, Zabol University, Zabol,Sistan, Baluchestan, Iran; bFaculty of Natural Resources and Marine Sciences, Department ofEnvironmental Sciences, Tarbiat Modares University, Nour, Mazandaran, Iran; cFaculty ofSciences, Department of Chemistry, Payam Nour University, Sari, Mazandaran, Iran; dFaculty ofNatural Resources, Department of Environmental Sciences, Science and Research University,Ahvaz, Khozestan, Iran

(Received 1 November 2008; final version received 8 January 2009)

Human breast milk samples, collected during April 2007 in Tabriz, Iran, wereanalyzed for organochlorine (OC) pesticides, such as dichlorodiphenyltrichlor-oethane (DDT) and its metabolites, hexachlorobenzene (HCB), �, �, and�-hexachlorocycloexane (HCH) isomers and six polychlorinated biphenyls(PCB) congeners (IUPAC Nos. 28, 52, 101, 138, 153, and 180). Organochlorinepesticides and OCB were both prevalent in the samples of human breast milk.Average concentrations of HCB, DDT, HCH, and PCB were 1020, 1930, 1660,and 690 ng g�1 lipid weight, respectively. A positive correlation was observedbetween concentration of OC in human breast milk and mothers age withprimiparae. Women having higher OC concentrations than multiparae womensuggests that these parameters influence the OC burden in lactating women. Theestimated tolerable daily intakes (TDIs) of HCH, HCB and PCB solely fromhuman breast milk were 100, 46, and 43% of samples, respectively, exceedingguideline thresholds given as TDI proposed by Health Canada. Although highdaily intakes may raise concerns for possible adverse effects of OC, women inTabriz are recommended to breastfeed due to the numerous advantages formother and child.

Keywords: human breast milk; organochlorine pesticide; polychlorinatedbiphenyls; Iran

Introduction

Despite the ban on persistent organochlorines (OC) in most of the developed nations sincethe early 1970s, their uses continued for agricultural and public health purposes in manydeveloping countries until very recently (Minh et al. 2004). Moreover, due to theirlipophilc properties and their resistance to biochemical degradation, these compoundstend to accumulate in fatty tissues and biomagnify, reaching harmful concentrations inorganisms situated at the high-end of the food chain, such as humans (Vallack et al. 1998).

Mothers are exposed to OC through food chain contamination and environmentalpollution. These chemicals enter the body through oral, dermal and inhalation routes and

*Corresponding author. Email: dahmardeh_behrooz@yahoo.com

ISSN 0277–2248 print/ISSN 1029–0486 online

� 2009 Taylor & Francis

DOI: 10.1080/02772240902732472

http://www.informaworld.com

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are absorbed and distributed to different organs (Sim and McNeil 1992). Organochlorinesare transferred from mother to fetus and newborns through placenta and breast milk.Human milk is a non-invasive way of monitoring humans for OC contamination (Kanjaet al. 1992; Schade and Heinzow 1998; Skaare, Tuveng, and Sande 1988).

Disruption of hormonal activity by some persistent OC, such as polychlorinatedbiphenyls (PCB), dichlorodiphenyl trichloroethane (DDT), and hexachlorocyclohexanes(HCH) was suggested (Cheek et al. 1999; Colborn, vom Saal, and Soto 1993; Kelce 1995;Vos et al. 2000). These facts raised public concern towards the adverse effects of OC onhuman health, especially for infants.

Over 27,000 tons of pesticides have been used in the Islamic Republic of Iran during2000–2001. Of the commonly used pesticides, 25 are banned in other countries and somehave been withdrawn from use in the Islamic Republic of Iran. Yet, farmers can easilyobtain them on the black market or via cross-border smuggling and continue to use themon rice, cotton and citrus as a national pesticide (Heidari 2003). Although all OC havebeen banned in Iran since the late 1990s, there is not enough information on stockpiles ofthese OC available, at present. However, due to the price, effectiveness and traditional use,possibly coupled with a lack of law enforcement, the use of these pesticides is continuing inIran. Furthermore, previous investigations demonstrated the occurrence of OC insturgeons and fish from Iran (Ebadi and Shokrzadeh 2006; Kajiwara et al. 2003).

Organochlorine pesticide (OCP) concentrations in human milk have been used toassess trends in OCP pollution since the early 1970s and in particular to evaluate thesuccess of the ban of OCP in numerous countries (Chao et al. 2006; Erdogrul et al. 2004;Jaraczewska et al. 2006; Smith 1999; Szyrwinska and Lulek 2006). Most studiesconsistently show peak concentrations of OCP in the 1970s or early 1980s witha subsequent decrease in the concentrations (Bates, Thomson and Garrett 2002;Konishi, Kuwabara, and Hori 2001; Newsome and Davies 1995; Schade and Heinzow1998). However, a few reports appeared pertaining to the levels of OCP in human milk(Cok et al. 1999; Hashemy-Tonkabony and Fateminoss 1977) or adipose tissue(Burgaz, Afkham, and Karakaya 1995; Hashemy-Tonkabony and Soleimani-Amiri1978) in Iran. They have reported that daily intake for OCP for most infants andpeople were above the acceptable daily intakes established by FAO/WHO. As a result, itremains unclear whether the decrease in OCP concentrations in human milk samplesobserved after the ban in the 1980s has continued or is approaching a steady state. Thisfact emphasizes the need for more detailed study on the accumulation of OCP inindividuals from a few locations of Iran.

In the present study, human breast milk samples were collected from Tabriz (Iran) todetermine the concentrations of persistent OC, such as PCB, DDT, HCH, andhexachlorobenzene (HCB). The aim of this study was to evaluate the degree ofcontamination in Iran in comparison to other countries and investigate whether alterationsin the levels of these contaminants have occurred since 1974. Therefore, the kineticaccumulation of OC was studied and potential risks for breast-fed infants were alsoevaluated.

Material and methods

Human milk collection

Human milk samples were obtained in April 2007 from the maternity unit of theTabriz State Hospital in Tabriz City (Figure 1). All mothers were living in the Tabriz area

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for at least seven years and participated voluntarily in the study. Milk samples were takenfrom one breast manually at the end of the feeding and between 6 and 58 days ofpostpartum. The milk samples were kept on ice immediately after collection, transferred tothe Tarbiat Modares University environment lab and preserved there at �20�C untilanalysis. The age of mothers ranged from 19 to 37 years. Each donor completeda questionnaire to provide personal information, such as weight, occupation, smoking,previous nursing time, dietary habits, and place of residence. All the donors werenon-smokers (Table 1).

Figure 1. Map showing sampling location.

Table 1. Characteristic of the women and babies in this study.

Primiparae Multiparae Total

N 14 16 30MotherAge (year) 25 (19–35) 30 (25–37) 27Weight (kg) 69 (53–89) 66 (48–95) 67.5Height (cm) 140 (160–168) 163 (150–170) 153Education 12 (9–16) 9 (5–16) 10Occupation Housewife: 85% Housewife: 90% Housewife: 87%

Others: 15% Others: 10% Others: 13%BabyAge (day) 27 (14–55) 29 (6–58) 28Weight (kg) 3.6 (2.7–4.8) 4 (2.8–5.9) 3.8Height (cm) 49 (30–57) 52 (46–60) 51Weight in birth 3.1 (2.7–3.9) 3.4 (2.5–4.20) 3.3

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Materials

The PCB standards (IUPAC Nos.: 28, 52, 101, 138, 153, 143 and 180), together with theOCP, �-, �-, �-, "-HCHs, p,p0-DDE, o,p0-DDE, p,p0-DDT, o,p0-DDT, p,p0-DDD, and HCBwere obtained from Dr Ehrenstorfer Laboratories (Augsburg, Germany).

The selected PCB congeners were detected in river water and cow’s milk in Iran(Bidkhori 1996; Esmaili-Sari, Dahmarde Behrooz, and Bahramifar 2008), and thereforethis study focused on measurements of these PCB congeners in human breast milk.Standards of o,p0-DDD were not available in our lab. Acetone, n-hexane, dichlor-omethane, diethyl ether, and iso-octane were of pesticide grade (Merck, Germany).Anhydrous sodium sulfate and silica gel (Merck) were used after heating at 120�C.

Analytical procedures

The determination of selected PCB and OCP in milk samples were performed as describedby Covaci, Hura, and Schepens (2001) and Erdogrul et al. (2004) with minormodifications. All samples were thawed and homogenized by shaking the milk for atleast 5min. An accurate amount of milk (1–6 g, depending on the sample availability) wasweighed and internal standards (15 ng PCB 143 and "-HCH) were added. After theaddition of 2mL of formic acid, the extraction was performed with 2� 12mL n-hexane/dichloromethane (5 : 1, v/v) by vortexing for 1.5min. The organic layer was then subjectedto clean-up on a cartridge containing 8 g silica, impregnated with concentrated sulfuricacid (44%, w/w). Complete elution of PCB and OCP was conducted with 15mL n-hexane,followed by 10mL dichloromethane. The final elute was concentrated with a rotaryevaporator and further under nitrogen to near dryness. The extract was solubilized in80 mL iso-octane and transferred to an injection vial.

Procedures for extraction of lipids from milk described by Johansen et al. (1994) wereused with minor modifications. Lipid determination was conducted on a separate aliquotof milk by extracting 1 g milk with 2� 3mL n-hexane : diethyl ether (1 : 1, v/v) for 2min.The organic layer was transferred to an aluminum dish and evaporated. Lipids weregravimetrically measured after keeping the dish at 105�C for 1 h.

Quantification

Gas chromatography (GC) analysis was performed using a Dani 1000 gas chromatographequipped with 63Ni electron capture detector. The GC system was equipped with a DB-5capillary column (60m� 0.25mm i.d., 0.25 mm film thickness, Macherey–Nagel). Heliumgas was used as the carrier gas at a flow rate of 2mLmin�1. The oven temperature wasinitially maintained at 100�C for 1min and was increased at a rate of 10�C min�1 to 240�Cand held for 1min. Then, oven temperature was programmed with a rate of 1�C min�1

until it reached 260�C for 1min and was increased at a rate of 20�C min�1 to 300�C,retained at this temperature for 10min while the injection port was operated in a splitlessinjection mode. The injection and detector temperature were set at 250�C and 300�C,respectively. An injection sample volume of 1 mL was used for liquid analysis.

Quality assurance

Multi-level calibration curves were created for the quantification, and linearity (r24 0.999)was achieved for the whole concentration range found in the samples. The recovery tests

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were performed to check the efficiency of the analysis method employed for thedetermination of OC in milk. Spiking was done at two levels, 50 and 500 ng g�1 lipid.A method blank sample was included in each batch of 10 samples. The results showed thatOC were below the detection limit in all blanks. Method limits of quantification (LOQ) forindividual PCB were between 0.03 and 0.09 ng g�1 lipid wt. and between 0.01 and 0.2 ng g�1

lipid wt for OCP (Table 2). The recovery rates of OCP and PCB were between 90 and 110%

(RSD5 10%).

Statistical analysis

A Kruskal–Wallis one-way analysis of variance, followed by a Mann–Whitney U test, wasused to evaluate the effect of parity on the OC concentrations. Spearmen’s rankcorrelation coefficients were used to test the correlations between the age of mother andOC concentrations. A p-value 50.05 was considered to indicate statistical significance.The statistical analysis was done by the SPSS software (Version 11.5). In this study, we didnot have outliers.

Results

The OC present at highest concentration were HCB (1020 ng g�1 lipid wt.), DDT(1930 ng g�1 lipid wt.), HCH (1660 ng g�1 lipid wt.), and PCB (690 ng g�1 lipid wt.)(Table 2). The detection frequency for all compounds is given in Figure 2 for OC andFigure 3 for PCB. OCP and PCB were the major contaminants in the samples from Tabriz,

Table 2. Concentration of OCP and PCB in human breast milk (ng g�1 lipid wt.) in Tabriz, Iran.a

Compound Primiparae (n¼ 14) Multiparae (n¼ 16) Totalb LOQ

% fat 1.4 (0.3–4.3) 1.7 (0.4–5) 1.6 (0.3–5)�-HCH 330 (n.d–1740) 370 (n.d–1530) 350 (n.d–1740) 0.01�-HCH 820 (n.d–5265) 660 (50–3400) 735 (n.d–5265) 0.1�-HCH 950 (n.d–5930) 250 (n.d–1140) 580 (n.d–5930) 0.2HCHs 2100 (300–13780) 1280 (80–4220) 1660 (78–13780)HCB 1500 (n.d–5980) 590 (n.d–6470) 1020 (n.d–6470) 0.1o,p0-DDT 310 (n.d–1380) 280 (n.d–2490) 295 (n.d–2490) 0.08p,p0-DDT 420 (n.d–2540) 180 (n.d–1800) 290 (n.d–2540) 0.2o,p0-DDE 55 (n.d–430) 120 (n.d–344) 90 (n.d–430) 0.1p,p0-DDE 1325 (50–7799) 880 (35–2840) 1090 (n.d–7780) 0.08p,p0-DDD 145 (n.d–930) 190 (n.d–1550) 170 (n.d–1550) 0.1DDTs 2260* (100–10340) 1640* (20–1290) 1930 (20–10340)PCB 28 20 (n.d–190) 20 (n.d–180) 20 (n.d–190) 0.09PCB 52 110 (n.d–1500) 40 (n.d–330) 70 (n.d–1500) 0.04PCB 101 150 (n.d–600) 190 (n.d–1135) 170 (n.d–1135) 0.03PCB 138 100 (n.d–570) 110 (n.d–310) 100 (n.d–570) 0.04PCB 153 100 (n.d–520) 150 (n.d–420) 120 (n.d–520) 0.05PCB 180 250 (n.d–1750) 150 (n.d–400) 200 (n.d–1750) 0.03PCBs 730 (n.d–1750) 650 (31–1378) 690 (n.d–1750)

Note: n.d – not detected; LOQ-Limit of quantification.aAverages were given.bConcentration from total mothers (primiparae and multiparae).*p5 0.05.

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Iran (Table 2) with p,p0-DDE being measured in all samples, and �-HCH having

a detection frequency of 90% (Figure 2). The distribution pattern in the studies by Cok

et al. (1999) and Burgaz, Afkham, and Karakaya (1995) in Iranian human breast milk

samples followed the order of DDT4HCH4HCB. Residue levels of DDT, HCH, and

HCB in agriculture were the most important source of OCP in human milk samples from

Iran (Cok et al. 1999). The mean ratio between DDE and DDT was 2. This value is low as

compared to that obtained in 1992 (5.6). This indicates that there is new input of DDT in

this region of Iran.�-HCH was the most prevalent HCH isomer with a mean of 735 ng g�1 lipid wt. The

mean concentration of �-HCH was 580 ng g�1 lipid wt., while the mean concentration of

�-HCH was 350 ng g�1 lipid wt. (Table 2). HCB was found in 14 human milk samples

(46%) with a mean concentration of 1020 ng g�1 lipid wt. PCB were measured in 29 of 30

samples. The most frequently detected PCB congener was PCB 180, while PCB 28 and

PCB 52 had the lowest detection frequency (Figure 3).

0

10

20

30

40

50

60

70

80

90

100

p,p′- DD

E

PCB

s

p,p′- DD

D

p,p′- DD

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HC

B

o,p′- DD

TFr

eque

ncy

of d

etec

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(%)

-HC

B

-HC

B

-HC

B

b g a

Figure 2. Detection frequency of OCs in human milk from Tabriz, Iran.

0102030405060708090

100

180 101 153 138 28 52

PCB No.

% P

CB

Figure 3. PCBs profile in human milk from Tabriz, Iran.

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Higher concentrations of OC were found in human breast milk of primiparae as

compared to the multiparae (Table 2). However, only the level of DDT was significantly

different between the two groups. Figure 4 shows the relationship between parity and

concentration of DDT in human breast milk from Tabriz city. Because of the potential

influence of parity on the levels of OC, the correlation between OC levels and mothers’ age

was examined, independently for primiparae and multiparae. In the case of multiparae, the

levels of all OC did not correlate with age, but levels of HCH, PCB and DDT in the

primiparae group were positively correlated with age. The correlation was significant only

for DDT (Figure 5).

Discussion

Contamination status

From 1979, restrictions were imposed in Iran against the use of DDT in agriculture, and

since 1983, its use has been prohibited. However, DDT has been allowed for malaria

control (Cok et al. 1999), but the high detection frequency of p,p0-DDT in the human milk

samples suggests that DDT has been used recently in the studied region. Moreover, the

DDE/DDT ratio found in the present study was lower than the ratio measured in samples

obtained in 1991 from the same region (Cok et al. 1999).Mean concentration levels of DDT in human milk samples were 1930 ng g�1 lipid,

while mean concentration levels of DDT in human milk samples in developing counties

such as Mexico were 4700 ng g�1 lipid (Waliszewski et al. 2001), Ukraine 2700 ng g�1 lipid

(Gladen et al. 1999) and in developed countries such as Australia 1200 ng g�1 lipid

(Quinsey, Donohue, and Ahokas 1995), Japan 430 ng g�1 lipid (Kunisue et al. 2004c),

Germany 240 ng g�1 lipid (Schade and Heinzow 1998) and the Czech Republic 1050 ng g�1

lipid (Schoula et al. 1996). Furthermore, the levels of DDT in the present study were lower

than the levels measured in Tabriz in 1991 (Cok et al. 1999) and Tehran (Hashemy-

Tonkabony and Fateminoss 1977) that were 2220 and 2880, respectively.Differences in the persistence and bioaccumulation potential among HCH isomers may

change their ratio throughout the food chain up to the excretion in human milk, resulting

Figure 4. Concentration of DDTs in maternal milk vs. the corresponding number of children inTabriz. The short horizontal bars indicate mean concentration in group and the stars (*) indicatesignificant difference ( p5 0.05).

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in the more persistent �-HCH being the predominant isomer in human milk (Szokolayet al. 1977). Among the investigated HCH isomers, �-HCH had the second highestdetection frequency (Figure 2). This insecticide has a shorter environmental half-lifecompared to most other OC, and it is relatively rapidly metabolized and excreted byorganisms (Walker 2001). Therefore, the presence of �-HCH in human milk samples inTabriz most likely reflects a recent exposure of mothers to lindane, which has replaced thetechnical HCH and which is currently used for agricultural purposes in this region. In thisstudy, the levels of HCH in human milk in Tabriz were 1660 ng g�1 lipid that were lowerthan those obtained in countries like India (Subramanian et al. 2007) and Kazakstan(Hooper et al. 1997), that were 4500 and 2300 ng g�1 lipid, respectively, but at higherconcentration levels in countries like Japan (Kunisue et al. 2004c), Germany 40 ng g�1 lipid(Schade and Heinzow 1998), Russia 810 ng g�1 lipid (Tsydenova et al. 2006). Additionally,the present HCH levels are higher than those previously found in Tabriz adipose tissue(770 ng g�1 lipid wt., Burgaz, Afkham, and Karakaya 1995) or in Tabriz human breastmilk (600 ng g�1 lipid wt.) (Cok et al. 1999). This suggests that there is a new input of HCHin the region. This is also supported by the fact that official reports indicate that HCH areused in agriculture and have replaced DDT. A study on pesticide residues in 18 rivers ofthe three Caspian coastal provinces of Iran conducted in 1997, showed several OCPbelonging to the Persistent Organic Pollutants (POP) convention list, while lindane wasfound in water at concentrations between 8 and 4000 ppb (Heidari 2003).

While concentration levels of HCB in human milk samples in countries like China(Kunisue et al. 2004a), Malaysia (Sudaryanto et al. 2005), India (Subramanian et al. 2007),Turkey (Cok et al. 1997), and Germany (Schade and Heinzow 1998) were 81, 11, 50,and 80 ng g�1 lipid, respectively, the concentration level of HCB in this study was100 ng g�1 lipid and, in fact, was 6–242 orders of magnitude higher than correspondinglevels in human milk in the above-mentioned countries. The concentration level of HCB

y= 77.783x – 1200.9 R2= 0.2115

0

400

800

1200

1600

2000

10 15 20 25 30 35 40Age of mothers (Years)

PC

Bs

conc

entr

atio

n(n

gg–1

lipi

d w

t.)

y= 16.971x + 1086.2 R2= 0.0012

0

2000

4000

6000

8000

10 20 30 40Age of mothers (Years)

HC

B c

once

ntra

tions

(ng

g–1 li

pid

wt)

y= 414.3x– 8038.2 R2= 0.4452

0

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10 15 20 25 30 35 40Age of mothers (Years)

DD

Ts

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t.)

p > 0.05

y= 531.49x – 10631

R2= 0.4115

0

4000

8000

12,000

16,000

10 20 30 40Age of mothers (Years)

HC

Hs

conc

entr

atio

ns(n

gg–1

lipi

d w

t.)

Figure 5. Relationship between age of mothers and OC concentration in primiparae.

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performed in this region in 1991 was 61 ng g�1 lipid (Cok et al. 1999), HCB levels detected

in the present study were 17-fold higher. This chemical has a high bioaccumulation

potential because of its high Kow and long half-life in biota (Naso et al. 2003). Most likely,the present findings of HCB are not only due to its previous use as a fungicide for seed

grain, but also to the fact that it is a by-product of various manufacturing and combustive

processes (Bailey 2001; Leoni and D’Arca 1976). Moreover, HCB may be formed bybiotransformation of �-HCH (Menzei 1986; Tobin 1986). Both usage of technical lindane

and industrial activities might help explain the source of HCB in human milk samples of

Tabriz residents.The present study is the first report on the accumulation of PCB in breast milk in

Iran. The mean value of sum six indicator PCB was 690 ng g�1 lipid wt. These PCBcongeners were frequently detected and the hepta-chlorinated congener PCB 180 was

predominant. Our previous study showed that higher concentrations of PCB 180 and PCB

138 might occur in this region (Bidkhori 1996). It seems that industrial plants,especially electrical industry, are a major source of PCB contamination in this region.

This congener is highly lipophilic, stable and accumulates in terrestrial ecosystems.

Moreover, because of the high degree of chlorination and the particular position of the

chlorine atoms on the biphenyl backbone (lack of unsubstituted adjacent meta and parapositions), this high-chlorinated biphenyls is refractory to metabolic attack and tend to be

more strongly bioaccumulate than the low-chlorinated ones (Walker 2001). Concentration

levels of PCB in the study of Schoula et al. (1996) in Czech Republic were 1160 ng g�1 lipid

compared to concentration levels PCB detected in this study. Levels of PCB indeveloping countries such as China (Kunisue et al. 2004a), India (Subramanian et al.

2007), Kazakstan (Hooper et al. 1997), and Malaysia (Sudaryanto et al. 2005), were 42, 34,

380 and 80 ng g�1 lipid, respectively, that was 3–20-fold lower than levels of PCB

in this study.

Specific accumulation according to parity and age

In lactating woman, several factors, such as number of children, age of mothers, foodintake preference, breast-feeding period, and several other external factors, have been

associated with the concentration of OC in human breast milk, (Albers et al. 1996; Czaja

et al. 1999; Harris, Woolridge and Hay 2001; Rogan et al. 1986). Concentration of OC in

human breast milk of mothers who fed only one child (primiparae) were higher than inthose mothers who had more than two children (multiparae). Although women who breast

fed may have had elevated serum OC levels prior to lactation, lactation might reduce these

levels by transferring the contaminants to the infant (Rogan et al. 1986). The influence of

parity on the concentrations of OC in breast milk agrees with other reports (Kunisue et al.2004b; Minh et al. 2004; Rogan et al. 1986; Soliman et al. 2003). Due to the lipophilic

nature of OC, lipid mobilization from adipose tissue to breast milk and excretion through

breast feeding is one of the possible explanations for the observed decreasing OCconcentrations with parity (Sudaryanto et al. 2006).

The observed positive correlation between the OC residue levels and the age of

primiparae mothers (Figure 5) may be explained by the assumption that the dietary food

intake is a major source of OC contaminants in the body (Albers et al. 1996) and that OC

are resistant to metabolism in the body and bioaccumulate with age. Similar results onincreasing OC levels in mother’s milk age in primiparae were previously reported (Albers

et al. 1996; Czaja et al. 1999; Minh et al. 2004; Sudaryato et al. 2006).

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Estimated daily intake of OCs by infant

Daily intakes by infants were calculated based on the average milk consumption of a 4 kg

infant is 699 g per day (Butte, Lopez-Alarcon, and Garza 2002). The tolerable daily intake

(TDI) value proposed by Health Canada was for PCB, HCH, HCB and DDTs 1, 0.3, 0.27

and 20 mg kg�1 body wt. per day, respectively (Oostdam et al. 1999). Mean daily intake for

infants in this study was 1.4 (0–12.8), 3.2 (0.48–15.5), 1.7 (0–12.8), and 2.8 (0.49–11.9) for

PCB, HCH, HCB, and DDT, respectively. The estimated daily values were compared with

TDI established by Health Canada. The estimated intakes of HCH were higher than the

guideline permissible value for all samples (100%). Estimated daily intakes of HCB

and PCB were higher than the corresponding guideline value limit, for 46 and 43% of

samples, respectively.These facts raised public concern about the adverse effects of these OC on human

health, especially for infants due to their susceptibility to environmental impacts.

Walkowiak et al. (2001) observed an association between postnatal exposure to PCB and

mental/motor development of children from 30 months onward. Furthermore, there is

evidence that �-HCH is also an environmental estrogen (Willett, Ulrich, and Hites 1998).

HCB was shown to be a reproductive toxin in animals (Alvarez et al. 2000) and was

also associated with significant immediate and chronic adverse health effects in humans

(Jarrell et al. 2002).Despite the fact that human breast milk is contaminated with POP, organizations,

such as the WHO and American Pediatric Association, still consider the benefits of

breast feeding against any possible risk acquired by exposure to these compounds in

breast milk and consistently recommend breast feeding (Abelsohn et al. 2002; Longnecker

and Rogan 2001). Through breast feeding, the infant obtains optimal nutrition,

immunological developmental, psychological, economic and practical advantages for

his/her health and development when compared to formulated feeding (Lonnerdal 2000;

Pronczuk, Moy, and Vallenas 2004; Uauy and Peirano 1999). It has been emphasized that

primary preventive measures to control and reduce the introduction of OC into the

environment were the most effective means to eliminate or at least reduce the levels of OC

in human milk.

Conclusions

The present study is the most recent comprehensive study on OC contamination in breast

milk from Iran. HCH, DDT and PCB were the most abundant contaminants detected in

the analyzed human milk samples. This seems to illustrate well the historical intensive use

of HCH and DDT and the on going use of PCB in Iran. Parity and age of the mothers

influenced the levels of OC in the human milk. Comparison of the obtained data with

results from other countries indicates that women in Tabriz have greater exposure to

relatively high levels of certain OC, namely HCH, PCB, and HCB. This finding

strengthens the necessity for further activities, in addition to the official ban, toward

a reduction and elimination of PCB and OCP in Iran. The estimated daily intake of HCH,

PCB and HCB by infants exceeded the corresponding Canada Health guidelines.

However, the presence of environmental chemicals in breast milk of women from Tabriz

should not discourage them from breastfeeding, as breast milk is critically important for

optimal child health, growth and development.

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Acknowledgments

We thank Hormoz Sohrabi who cooperated in the statistical analysis of the data. We also thankMs. S. Dahmardeh Behrooz for language assistance. We thank Ms. Khandan Shiri for collection ofhuman milk samples from Tabriz. We would like to thank Dr. Adrian Covaci (University ofAntwerp, Belgium) for critically reviewing this article.

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