~ Pergamon Chemosphere, Vol. 38, No. 5, pp. 995-1004, 1999 © 1998 Elsevier Seien~ Ltd. All rights reserved

0045-6535/99/$ - see front matter

PII: S0045-6535(98)00359-2

PCDD/F CONCENTRATIONS IN MILK OF NONOCCUPATIONALLY EXPOSED

WOMEN LIVING IN SOUTHERN CATALONIA, SPAIN

M. Schuhmacher 1, J.L. Domingo 2, J.M. Llobet 2, H. Kiviranta 3 and T. Vartiainen 3

1Environmental Engineering Laboratory, "Rovira i Virgili" University, 43007 Tarragona, Spain

2Laboratory of Toxicology and Environmental Health, "Rovira i Virgili" University,

San Lorenzo 21, 43201 Reus, Spain

3Division of Environmental Health, National Public Health Institute, 70701 Kuopio, Finland

(Received in Germany 19 May 1998; accepted 22 June 1998)

ABSTRACT

Concentrations of 17 toxic 2,3,7,8-substituted polychlorinateddibenzo-p-dioxins(PCDDs)

and polychlorinated dibenzofurans (PCDFs) have been determined in pooled samples of breast

milk from 15 mothers living in two residential areas (Tarragona downtown and an industrial

area) in the vicinity of a new hazardous waste incinerator (HWl), now under construction in

Tarragona (southern Catalonia, Spain). PCDD/Fs in human milk samples ranged between 5.9 and

17.1 pg I-TEQ/g fat (162-498 pg I-TEQ/I), with a mean value of 11.8 pg I-TEQ/g fat (310.8 pg

I-TEQ/I). The percentages of fat ranged between 1.53 and 3.52. Although PCDD/F levels in milk

from mothers living in the industrial area were found to be slightly higher than those observed

in women living Tarragona downtown, most differences did not reach the level of statistical

significance. In general terms, PCDD/F concentrations in human milk (pooled) samples of

mothers living in the area of Tarragona are similar or lower than those previously reported for

most industrialized countries. ©1998 Elsevier Science Ltd. All rights reserved

Key-words: human milk, PCDD, PCDF, I-TEO, Tarragona (Spain).

INTRODUCTION

Concerns about hazardous waste and unplanned releases of hazardous substances into

the environment are wide ranging [1-3]. Because of the effective volume reduction and chemical

995

996

toxicity destruction capabilities, incineration is often preferred to hazardous waste treatment or

disposal alternatives. However, the public image of incineration has now deteriorated in many

countries to the point where planned hazardous waste incinerators (HWl) are frequently rejected

by local populations, even when incineration can be one of the most cost-effective and

environmentally acceptable treatment option [4].

In recent years, the public health impact associated with stack emissions from HWl have

become a major concern. Although most evaluations on stack emissions have focused on three

classes of compounds: metals, semivolatile, and volatile compounds [5], without doubt, the

greatest amount of scientific and public attention has been given to one class of incineration

combustion byproducts: the polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated

dibenzofurans (PCDFs) [4-6].

A new hazardous waste incinerator is being constructed in the area of Tarragona

(southern Catalonia, Spain). Since the possible adverse effects of waste incinerators continue

being a subject of controversy and worry, only a wide knowledge of the impact of these

facilities could reasonably help to reduce the strong public opposition to waste incinerators. An

assessment of the future impact of the new HWl on public health requires a wide information

about the human baseline burden for substances such as metals and PCDD/Fs, which are

typically emitted by these facilities.

Because PCDDs and PCDFs are lipid soluble, milk and adipose tissue, together with

plasma, are frequently used as biomonitors of environmental exposure of humans to these

organic pollutants [7,8]. PCDD/Fs have been detected in milk from nonoccupationally exposed

women at relatively high levels. Since the bioavailability of PCDD/Fs from breast milk is high,

breast-fed infants can be potentially exposed to these highly toxic compounds. In relation to it,

PCDD/F concentrations in breast milk were reported to decrease with duration of lactation and

with the number of breast-fed children [7,8].

Recently, we determined the levels of a series of metals in hair, blood, and autopsy

tissues from subjects living in the area of the new HWl [9-1 1]. Moreover, the concentrations

of PCDD/Fs in plasma of individuals living in this area were also measured [12]. The purpose

of the present study was to determine the levels of PCDD/Fs in breast milk from women living

in the area under potential influence of the HWI.

EXPERIMENTAL

Samples

Fifteen pooled milk samples were obtained during 1996 from mothers living in the area of

Tarragona. Six of them corresponded to women living in the vicinity of an industrial

(petrochemical) zone, while the remaining 9 were obtained from women living downtown (urban

997

area). A questionnaire completed by the participants prior to the study did not indicate any

occupational exposure to PCDD/Fs. Questions about the present use of medication were also

computed. Since PCDD/F levels in human milk might be affected by a number of factors such

as dietary habits, age and body weight of the mothers, number of children, total period of

breast-feeding, as well as the percentage of lipid content in the milk [13-15], in order to reduce

the potential influence of these factors, all the participants in the study were primiparae

mothers, between 25-35 years, who had lived in the area of Tarragona for at least the last 5

years. All mothers had a single child and infants were exclusively breast-feeding. Both, mother

and child, were apparently healthy, and the pregnancy had been normal. The time elapsed

between delivery and sampling was between 2 weeks and 2 months, as recommended by

WHO/EURO [16].

PCDD/F analyses

PCDD/F analyses in human milk were performed in pooled samples composed of a varying

amount (50-100 ml) of individual samples. Pooled samples were homogenized and freeze-dried.

Fat was extracted with a mixture of diethyl ether and hexane after addition of sodium oxalate

and ethanol. The solvent was exchanged to hexane and the fat content was determined. After

addition of internal standards, an aliquot of 1-1.5 g of fat was purified over a silica gel column.

The sample was fractionated using activated carbon column with celite and cleaned with

activated alumina columns. 13C-labeled internal standards were used for quantification. A total

of 16 congeners of PCDD/Fs (8ED-998 tetra-octa chlorodioxin standard solution and EF-999

tetra-octa chlorofuran standard solution, Cambro Scientific) were used to quantify PCDD/Fs. A

gas-chromatography (Hewlett-Packard 5890) with fused silica capillary column, J&W Scientific

DBDXN (60 m, ID 0.25 mm, film 0.1 5 pm) using SIR-mode and resolution of 10000, was used

for PCDD/F analyses. Limits of quantification were between 0.05 to 0.5 pg/g fat.

RESULTS AND DISCUSSION

In the current study, 15 pooled samples of human milk were analyzed. The percentages

of fat ranged between 1.53 and 3.52 (Table 1). The levels (I-TEQ) of PCDD/Fs in the milk

samples are shown in Table 1. PCDD/F concentrations ranged between 5.9 and 17.1 pg I-TEQ/g

fat (162-498 pg I-TEQ/I), with a mean value of 11.8 pg I-TEQ/g fat (310.8 pg I-TEQ/I). In

comparison with the results of previous studies in Spanish populations, the current data are

similar to those obtained by Gonzalez et al. [17] in milk from women living in Madrid. However,

they are lower than the levels of PCDD/Fs found in women of the Basque Country (NE Spain)

[18].

998

Table 1: Individual levels of PCDD/Fs and percentages of fat in pooled samples of breast milk

from mothers living in the area of Tarragona (southern Catalonia, Spain)

Sample code pg I-TEQ/I pg I-TEQ/g fat Fat %

1 215 ~14.0 1.53

2 162 5.9 2.73

3 212 11.6 1.83

4 319 10.8 2.96

5 498 17.1 2.91

6 414 13.8 3.00

7 282 8.0 3.52

8 362 12.3 2.94

9 443 14.1 3.14

10 372 13.7 2.71

11 282 11.7 2.41

12 231 9.3 2.47

13 328 11.6 2.83

14 263 12.3 2.14

15 272 11.1 2.42

Recently, PCDD/F concentrations in plasma from the inhabitants of the area of Tarragona

were also determined [12]. PCDD/F levels in plasma resulted to be a 42% higher in people from

Tarragona than those found in the population of Madrid [19]. The differences between our

results in human plasma and milk, and those obtained in Madrid can be due to the different

methods of lipid extraction, which could in turn result in different analytical results [20].

Moreover, the effect of factors such the age, or very especially the dietary habits, may also play

an important role in PCDD/F accumulation in fat tissues [21].

The levels of PCDD/F congeners, as well as the calculated I-TEQs for the 15 pooled milk

samples are given in Table 2. It can be seen that PCDD levels diminished with the decreasing

degree of chlorination. 2,3,7,8-TCDD was identified in all samples with a mean value of 1.04

pg/g fat. A different pattern was found for PCDFs. For these compounds, 2,3,4,7,8-PeCDF was

the predominant congener, followed by 1,2,3,4,7,8-HxCDF. In most samples, 1,2,3,7,8,9-

HxCDF and OCDF concentrations were under their respective detection limits.

Figure 1 shows the congener profile of PCDD/Fs in milk samples of women living in the

vicinity of the new HWl and clasiffied according to two different residential areas (Tarragona

999

downtown and an industrial area). PCDD/F levels in milk from mothers living in the industrial

area were found to be slightly higher than those observed in women living downtown (mean

values: 12.7 and 11.2 pg/g fat, respectively). However, the differences did not reach the level

of statistical significance with the exception of 1,2,3,7,8,9-HxCDD whose concentrations were

significantly higher (p < 0.05) in milk from mothers living in the industrial area than in those

living downtown. PCDD/F concentrations in human breast milk levels have been generally found

to be higher in polluted industrial areas.

Table 2: Levels (pg/g fat) of PCDD/F congeners in pooled samples of breast milk from mothers

living in the Tarragona area (southern Catalonia, Spain)

CONGENER Mean SD Minimum Maximum

2,3,7,8-TCDD 1.04 0.07 0.59 1.53

1,2,3,7,8-PeCDD 4.02 0.92 1.93 5.62

1,2,3,4,7,8-HxCDD 2.84 0.57 1.27 3.32

1,2,3,6,7,8-HxCDD 27.88 1.54 15.10 38.00

1,2,3,7,8,9-HxCDD 4.55 1.34 2.26 7.53

1,2,3,4,6,7,8-HpCDD 38.26 24.54 9.36 118.00

OCDD 145.67 40.20 53.00 234.00

2,3,7,8-TCDF 0.68 0.22 0.33 1.11

1,2,3,7,8-PeCDF 0.31 O. 12 O. 12 0.61

2,3,4,7,8-PeCDF 7.95 2.11 3.85 11.00

1,2,3,4,7,8-HxCDF 3.06 0.50 1.91 3.87

1,2,3,6,7,8-HxCDF 2.49 0.48 1.49 3.44

1,2,3,7,8,9-HxCDF 0.04 0.02 ND 0.08

2,3,4,6,7,8-HxCDF 1.00 0.28 0.36 1.47

1,2,3,4,6,7,8-HpCDF 2.00 0.61 0.90 3.19

1,2,3,4,7,8,9-HpCDF O. 14 0.07 0.02 0.30

OCDF 0.34 0.16 ND 0.66

Total I-TEQ (PCDD/Fs) 11.82 2.71 5.93 17.10

Fat content (%) 2.63 0.52 1.53 3.52

Table 3 shows a summary of results concerning a number of recent studies about PCDD/F

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concentrations in human milk from a number of different countries and regions. While PCDD/F

levels (I-TEQ) in milk from women living in the area of Tarragona are rather similar to those

found in Norway [22], Germany [23], or Madrid (Spain) [17], they are lower than those obtained

in Finland [24], France [17], The Netherlands [25], USA [26], Jordan [27] or the Basque

Country (Spain) [18]. By contrast, they are higher than those found in Israel and Palestina [28].

In any case, it should be taken into account that data from human milk are rather difficult to

compare due to a series of variables that can contribute to the PCDD/F content in this fluid [24].

Table 3: Summary of recent studies on PCDD/F levels (pg I-TEQ/g fat) in milk from women

living in a number of different countries

Place No. of PCDD/Fs Year of Reference

samples collection

Groningen/ 168

Rotterdam, NL

Norway, 3 cities 10

Lithuania, 3 cities 12

Paris, France

Madrid, Spain

Schwandorf, FRG

Jordan,

5 different towns

Basque Country, 29

Spain

Binghamton, USA 9

Germany 38

Bethelem, 5

Palestina

Jerusalem, Israel 1

Southern and eastern

Finland 167

Tarragona, Spain 15

30.0 ....

(11.1-76.4)

10.4 ....

14.8 ....

15 20.1 1990

13.3 13 1990

7 12.4 (6-19) 1993

6 35 1994

(0.26-60.32)

19.4-25.5 1990-95

22 1995-97

16 1995

6.46 1996

10.19 1996

26.3/20.1 1987

(14.7-96.3)

11.8 1996

(5.9-17.1)

Tuinstra et al. 1994 [25]

Becher et al. 1995 [22]

Becher et al. 1995 [22]

Gonzalez et al. 1996 [ 17]

Gonzalez et al. 1996 [17]

Deml et al. 1996 [23]

Alawi et al. 1996 [27]

CAPV 1997 [ 18]

Schecter et al. 1997a [26]

Schecter et al. 1997a [26]

Schecter et al. 1997b [28]

Schecter et al. 1997b [28]

Vartiainen et al. 1997 [24]

This study

The average PCDD/F intake of an infant of 5 kg ingesting 700 ml/day of breast milk

would be 43.4 pg I-TEQ/kg/day. This value is 4.3 times greater than the tolerable intake of 10

pg I-TEQ/kg/day established by the WHO for the general population. However, because of the

]O02

relatively short nursing period and the rapidly increasing amount of adipose tissue of the infant,

it has been pointed out that the tolerable intake values established for lifetime exposures would

not be applicable for breast-fed infants [16]. In relation to PCCD/F intake by infants, McLachlan

[29] reported that for almost all PCDD/F congeners, over 90% of the compounds ingested by

a breast-fed infant were absorbed, whereas Ayotte et al. [30] estimated through a toxicokinetic

model that breast feeding strongly influences body burden during childhood. However, Kreuzer

and coworkers [31] have showed that the half-life of TCDD in humans is age dependent. The

lowest half-life is 1.5 years for newborns and about 10 years at the age of 40 years.

In summary, the levels of PCDD/Fs in human milk (pooled) samples of mothers living in

the area of Tarragona are similar or lower than those previously reported for most industrialized

countries. The dietary habits, based on the "mediterranean diet", could be one of the main

reasons that would explain the relatively low PCDD/F levels found in this study. On the other

hand, although the tolerable daily intake of PCDD/Fs (10 pg I-TEQ/kg/day), which is based on

lifetime intake, should not be applied to the relative short nursing period, the current results are

far above these levels. Despite the fact that, to date, no adverse health effects in babies could

be directly related to background levels of PCDD/Fs in human milk, it seems quite reasonable

that all efforts should be undertaken to minimize the emission of these pollutants and to shut

down known sources to achieve a reduction of the human body burden. The present results

should be useful for future assessments of the impact on public health of the HWl now under

construction.

ACKNOWLEDGEMENTS

This research was supported financially by the Generalitat de Catalunya (Spain): Junta

de Residus, Department of Environment.

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