Thomas M Krauss

Ana Maria C B Braga

Global Monitoring Plan

Brazilian Contribution

POPs in Human Milk

Fundação Oswaldo Cruz Escola Nacional de Saúde Pública Centro de Estudos da Saúde do Trabalhador e Ecologia Humana

Analytical Capacity Sample Preparation: Freeze dryer, Centrifuge

Extraction: Classic Soxhlet Extractor, Liquid-Liquid

Cleanup: Open Columns (Biobeads, Silica, Florisil, etc.)

Instrumental: HRGC/LRMS and HRGC/µ-ECD

Matrixes:

Soil (PCDD/F, Indicator PCBs, Pesticides, PAHs)

Sediment (PCDD/F, Indicator PCBs, Pesticides, PAHs)

Air Particles (PCDD/F, Indicator PCBs, PAHs)

Human Milk (Indicator PCBs, Pesticides)

Human Blood (Indicator PCBs, Pesticides)

Personal: At the moment only one Chemist for POPs

GMP Phase 1

Second Study 2011 - 2014

Fifth Study of Persistent

Organic Pollutants in Human

Milk coordinated by WHO in

collaboration with UNEP

First study was conducted in 2002 attending the call for participation in the

Third Round of Exposure Studies – PCDD/Fs and PCBs in Human Milk

coordinated by WHO/European Center for Environment and Health

GMP Phase 1

Contact by the Department for Climate Change and

Environmental Quality of the Ministry of Environment

(MMA), focal point of the Stockholm Convention in

Brazil.

Project elaboration for monitoring of POPs in Human

Milk in order to fulfill the obligations of the convention

and introduce Brazil in the GMP.

Approval of the project e financial support by the

ministry – Collaboration between FIOCRUZ and MMA.

National Coordination: Ana Maria C. B. Braga e Thomas

M. Krauss.

GMP Phase 1 Elaboration of the national protocol for human milk sampling

based on the WHO guidelines (2007) and the protocol of the first

study.

National Protocol used as Base for Standard Operating

Procedure for human milk sampling in the Grulac region.

http://www.ccbasilea-crestocolmo.org.uy/wp-

content/uploads/2010/11/SOP-Regional-de-Muestreo-de-

COP-en-Leche-materna.pdf

http://www.ccbasilea-crestocolmo.org.uy/wp-

content/uploads/2010/11/ing1.final_.SOP-POP-Regional-

Sampling-Breast-Milk-1.pdf

Sampling Locations

First Brazilian Study – Third Round of Exposure Studies

Sampling Locations Second Study (2011 – 2013) – GMP1

Grande Região 1 Grande Região 2 Grande Região 3

Rio Branco/Acre (3) Maceió/Alagoas (9) Brasília/ Distrito Federal

Macapá/Amapá Salvador/Bahia (10) Goiás/Goiânia (11)

Manaus/Amazônia Fortaleza/Ceará Belo Horizonte/Mina Gerais (12)

Belém/Pará (5) São Luis/Maranhão (6) Vitória/Espírito Santo

Porto Velho/Rondônia (2) João Pessoa/Paraíba (7) Rio de Janeiro/ Rio de Janeiro (13)

Boa Vista/Roraima (4) Recife/Pernambuco (8) São Paulo/São Paulo (14)

Palmas/Tocantins Teresina/Piauí Curitiba/Paraná

Cuiabá/Mato Grosso (1) Natal/Rio Grande do Norte Porto Alegre/Rio Grande do Sul

Campo Grande/Mato Grosso do Sul Aracaju/Sergipe Florianópolis/Santa Catarina (15)

Sampling Strategy - GMP Phase 1 Volunteer collaboration of human milk banks which are part of the

Brazilian Network of Human Milk Banks

Seeking of donors (eligibility criteria)

Application of the questionnaire

Human milk sampling

Human milk banks have the following vantages:

Sampling is done by professional that are accustomed with human

milk sampling handling of the samples;

Milk banks have the infrastructure for adequate storage of the

samples;

Professional are aware about the importance of breastfeeding.

All participating banks were visited and the staff trained for the special

purpose of the study. Delivery of all material (prepared sample bottles,

eligibility guide, consent form, questionnaire and a manual with

detailed instructions.

Type, number and volume of samples

150 individual samples collected – 100 mL

15 local pool samples - 350 mL (+ 150 mL reserve)

analysis of dioxins, PCBs, POPs pesticides and

PBDEs at the reference laboratory

3 regional pool samples - 750 mL analysis of all

POPs at the reference laboratory

150 individual samples - 35 mL analysis of DDT

and metabolites, and indicator PCBs at the

laboratory of FIOCRUZ.

Sample Shipment

Substâncias analisadas Compound/Compound group Regional pool (n = 3) Local pool (n = 15) Individual sample (n = 160)

Antigos POPs

PCDDs SIM SIM

PCDFs SIM SIM

dl-PCBs SIM SIM

PCBs indicadores SIM SIM SIM

DDT SIM SIM SIM

Clordano SIM SIM

Heptacloro SIM SIM

Aldrin SIM SIM

Dieldrin SIM SIM

Endrin SIM SIM

Toxafeno SIM SIM

Hexaclorobenzeno (HCB) SIM SIM

Mirex SIM SIM

Novos POPs

Hexaclorociclohexano (HCH) SIM SIM

Clordecona SIM

Hexabromobifenila SIM SIM

Pentaclorobenzeno SIM SIM

Endosulfam SIM SIM

Ácido perfluorooctanosulfônico (PFOS) SIM SIM

Polibromodifenil éteres (PBDEs) SIM SIM

Candidatos POPs

Parafinas cloradas de cadeia curta (SCCPs) SIM

Hexabromociclododecano (HBCD) SIM

Outras substâncias não POPs

Polibromodibenzo-p-dioxinas (PBDDs) SIM

Polibromodibenzofuranos (PBDFs) SIM

Dibenzo-p-dioxinas polihalogenadas (PXDDs) SIM

Dibenzofuranos polihalogenados (PXDFs)d SIM

Substâncias perfluoradas (PFCs) SIM SIM

Results – PCDDs/PCDFs e dl-PCBs

PCDD/Fs and dl-PCBs Medians - First study 2002

0

5

10

15

20

25

30

35

pg

EQT-

OM

S 19

98/g

de

go

rdu

ra

País

PCDDs/PCDFs dl-PCBs

Results– Indicator PCBs

Indicador PCBs Medians - First study 2002

0

50

100

150

200

250

300

350

400

450

500

550

ng/

g d

e g

ord

ura

País

PCBs indicadores

Results – PCDD/Fs Average levels of PCDD/Fs found in human milk of de 23 countries in ther period

of de 2008/2009 compared to average level found in Brazil in 2012

Average Brasil 2012:

2,38 pg EQT-OMS2005/g de gordura

2,73 pg EQT-OMS1998/g de gordura

0

5

10

15

20

25

pg

EQT-

OM

S 19

98/g

de

go

rdu

ra

País

PCDDs/PCDFs

Results – dl-PCBs Average levels of dl-PCBs found in human milk of de 23 countries in ther period of

de 2008/2009 compared to average level found in Brazil in 2012

Average Brasil 2012:

0,82 pg EQT-OMS2005/g de gordura

1,14 pg EQT-OMS1998/g de gordura

0

2

4

6

8

10

12

14

pg

EQT-

OM

S 19

98/g

de

go

rdu

ra

País

dl-PCBs

Results – IndicatorPCBs

Average levels of indicator PCBs found in human milk of de 23 countries in ther

period of de 2008/2009 compared to average level found in Brazil in 2012

0

10

20

30

40

50

60

70

80

90n

g/g

de

go

rdu

ra

País

PCBs indicadores

Average = 5,03 ng/g de gordura

GMP 1 Benefits and Problems

Stimulating the execution of national projects

Fincancial support – unfortunately low

Support with consumables, eg HT-8 for PCBs analysis

Proficiency test

Complicated bureaucratic procedures for establishing

formal collaborations – international and national level.

Long sampling times – other sampling strategy?

Restriction of donors maximum age

Challenges and expectations for the GMP-2 project

Increase of sample number – sampling in all state capitals of

Brazil including the Federal District – at least 270 individual

samples – maintain three regional pools with 90 individual

samples.

Involve stakeholders of the Ministry of Health on state or

municipality level – commitment of a local staff for seeking

donors – only sampling and storage at human milk banks.

Increase laboratorial capacity

Equipment for analysis of dioxins and other POPs of

complex analysis HRGC/HRMS and HRGC/MS/MS.

Contract technicians

Acknowledgement

The study coordinators are grateful to all the coordinators of the participating banks of

the Brazilian Network of Human Milk Banks and their staffs for their voluntary work

and great effort to identify donors and collect human milk samples. Without the support

of these banks it would not have been possible to collect samples all over the national

territory with such a large dimension.

In particular, we thank all the donors for the availability to answer the questionnaire and

the kindness to grant the collection of their milk for this research.

Contacts

Ana Maria C. B. Braga - anabraga@ensp.fiocruz.br

Thomas M. Krauss - thomas@ensp.fiocruz.br