EPA-600/1-76-032
September 1976
Environmental Health Effects Research Series
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into five series. These five broad
categories were established to facilitate further development and application of
environmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The five series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
This report has been assigned to the ENVIRONMENTAL PROTECTION
TECHNOLOGY series. This series describes research performed to develop and
demonstrate instrumentation, equipment, and methodology to repair or prevent
environmental degradation from point and non-point sources of pollution. This
work provides the new or improved technology required for the control and
treatment of pollution sources to meet environmental quality standards.
EPA REVIEW NOTICE
This report has been reviewed by the U.S. Environmental
Protection Agency, and approved for publication. Approval
does not signify that the contents necessarily reflect the
views and policy of the Agency, nor does mention of trade
names or commercial products constitute endorsement or
recommendation for use.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161
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EPA-600/1-76-032
September 1976
DDT LEVELS IN MILK OF RURAL INDIGENT BLACKS
By
Bennie T. Woodard, Bruce B. Ferguson
and
David J. Wilson
Meharry Medical College
MCH/FP Center
Box 69A
Nashville, TN 37208
Grant No. R802500
Project Officer
Dr. Ronald L. Baron
Environmental Toxicology Division
Health Effects Research Laboratory
Research Triangle Park, N.C. 27711
U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
HEALTH EFFECTS RESEARCH LABORATORY
RESEARCH TRIANGLE PARK, N.C. 27711
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DISCLAIMER
This report has been reviewed by the Health Effects Research
Laboratory, U.S. Environmental Protection Agency, and approved for
publication. Approval does not signify that the contents necessarily
reflect the views and policies of the U.S. Environmental Protection
Agency, nor does mention of trade names or commercial products
constitute endorsement or recommendation for use.
11
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FOREWORD
The many benefits of our modern, developing, industrial society are
accompanied by certain hazards. Careful assessment of the relative risk
of existing and new man-made environmental hazards is necessary for the
establishment of sound regulatory policy. These regulations serve to
enhance the quality of our environment in order to promote the public
health and welfare and the productive capacity of our Nation's population.
The Health Effects Research Laboratory, Research Triangle Park
conducts a coordinated environmental health research program in toxicology,
epidemiology, and clinical studies using human volunteer subjects. These
studies address problems in air pollution, non-ionizing radiation,
environmental carcinogenesis and the toxicology of pesticides as well as
other chemical pollutants. The Laboratory develops and revises air quality
criteria documents on pollutants for which national ambient air quality
standards exist or are proposed, provides the data for registration of new
pesticides or proposed suspension of those already in use, conducts research
on hazardous and toxic materials, and is preparing the health basis for
non-ionizing radiation standards. Direct support to the regulatory function
of the Agency is provided in the form of expert testimony and preparation of
affidavits as well as expert advice to the Administrator to assure the
adequacy of health care and surveillance of persons having suffered imminent
and substantial endangerment of their health.
Although the general use of DDT has been removed in the United States,
residues of this previously used and widely distributed pesticide persist
in the environment. This study was undertaken in an effort to assess the
level of contamination of humans in a previously high-use area and to
attempt to assess the potential for adverse human health effects.
H. Knelson, M.D.
.Director,
Health Effects Research Laboratory
in
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ABSTRACT
Human milk samples from low-income blacks residing in rural Miss-
issippi and Arkansas and middle-class whites residing in metropolitan
Nashville, Tennessee, were analyzed for DDT and its metabolites. The
mean total DDT (DDE + DDT) whole milk concentration of 38 samples from in-
digent blacks between April and September, 1974 was 447 ppb (range 59 to
1900 ppb) and the mean of the 14 samples from Nashvillians during the same
period was 75 ppb (range 15 to 133 ppb) . Seven samples from the black
population in June-September 1975 contained a mean total DDT of 323 ppb
(range 185-721 ppb).
This statistically significant difference in. the DDT concentrations in
the black and white populations indicates that the indigent blacks are still
highly contaminated with pesticides even though the general use of DDT has
been banned. Due to the limited amount of information from the donors,
no correlation could be made between the DDT concentration and any factors
other than race or socioeconomic group.
IV
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CONTENTS
Page
Abstract iv
List of Tables and Figures vi
SECTION I Conclusions 1
SECTION II Recommendations 2
SECTION III Introduction 3
SECTION IV Methods and Materials 6
SECTION V Results 9
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LIST OF TABLES AND FIGURES
Table 1 Summary of DDT Concentrations in Human Milk 4
Table 2 Pesticide Recovery Studies 8
Table 3 Pesticide Recovery Reproducibility 8
Table 4 DDT Levels in Human Milk - Present Study 1°
Figure 1 Distribution of Total DDT Levels in the Milk of Black Rural and
White Urban Women (1974) 12
VI
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SECTION I
CONCLUSIONS
The total DDT (DDT + DDE) levels in the milk of low income rural black
women in cotton-growing areas were found to be six times higher than those
found in middle class urban whites. The mean total DDT level for the blacks
(38 samples) in 1974 was 447 ppb (range 59 to 1900) in the whole milk; for the
whites in the same period it was 75 ppb (range 15 to 133). Seven samples
from the blacks taken in 1975 yielded a mean of 323 ppb (range 185 to 721).
These results indicate that low-income rural blacks in cotton growing
areas are still highly contaminated with DDT and its metabolites, although
apparently the levels are now decreasing. Due to the failure of most of
the donors to complete a questionnaire, no correlation could be made between
the DDT concentration and diet, age of child, home pesticide use, or distance
of residence from fields.
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SECTION II
RECOMMENDATIONS
Although the general use of DDT has been banned,32 this study indicates
a population that is obviously still highly contaminated with pesticides.
It is necessary to reiterate here that we have found no reported cases in the
literature of breast-fed infants being harmed by the DDT, but these low income
blacks from agricultural areas appear to be the population to study to deter-
mine if infants are in fact harmed by the DDT in human milk. We urge other
researchers to systematically study this group of women (whose milk contained
an average DDT concentration of almost a factor of ten greater than the WHO
limit for cow's milk) and attempt to determine if in fact infants are affected
by DDT in human milk. Our results alone, xv'ithout such clinical studies,
can not be used to justify either more or less stringent regulation of the
use of DDT, nor do they justify advising these black mothers not to nurse
their infants. Our results do justify, however, clinical study of these in-
fants which are being exposed to quantities of DDT greatly in excess of the
maximum admissible limit recommended by the World Health Organization.
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SECTION III
INTRODUCTION
The concentrations of DDT and its metabolites in human milk have been of
interest not only to mothers planning to breast feed their infants but also
to the general public.1 Table I summarizes the DDT concentrations found in
human milk by researchers the world over. The results presented in Table I
indicate that human milk generally contains DDT concentrations in excess of
the maximum set for cow's milk (50 ppb) by the WHO;27 the medical significance
of this is unknown.
The scientific and medical literature contains no confirmed cases of
infants suffering damage from the DDT in milk, but Fahim and coworkers28 have
reported increased mortality of nursing rats whose mothers were fed DDT.
Hayes,29 on the other hand, reported that adult humans given oral DDT doses
550 times greater than the average daily intake for 21.5 months showed no
definite clinical or laboratory evidence of damage for the five years over
which they were examined. Any damage to the human might well be expected to
be found in the susceptible infant rather than the adult, however.
Many low-income black families in the rural southern and southeastern
United States are exposed to somewhat different environmental factors than
other Americans. On numerous occasions, workers have been seen chopping cot-
ton in fields while the crop was being sprayed or dusted with pesticides
from airplanes. Many of the homes inhabited by these black families are
located within the confines of cotton fields which are sprayed. Windows,
doors, and cracks in the walls of these homes presumably allow the spray to
spread within the dwelling, where both food and water may be contaminated.
In 1970 alone, nearly a billion pounds of some 900 registered pesticides
were applied (more than 50 percent for farm use) throughout the United
States. ° Although regional statistics are unavailable, a 1964 survey by the
U.S. Department of Agriculture indicated that the cotton market accounted for
70 percent of the DDT used on farms.31 Even though a ban on the general use
of DDT took effect on January 1, 1973,32 a persistent pesticide such as DDT
would not immediately be removed from the environment.
Blacks in rural poverty areas of the South who have been exposed to high
pesticide concentrations for years would be expected to store above average
amounts of DDT and its metabolites in the adipose tissue due to the high
solubility of these compounds in fats and their relative insolubility in
aqueous solutions. Since DDT and its metabolites are excreted in the milk,
infants nursing black mothers residing in these areas would be logical
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candidates for pesticide related damage. Preliminary data on the DDT concen-
tration in human milk from women residing in these areas are here compared
with data of DDT concentrations in milk from women residing in less exposed
surburban areas.
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SECTION IV
METHODS AND MATERIALS
Samples of human milk were obtained froui black donors from rural poverty
areas in Bolivar County, Mississippi, and Lee County, Arkansas. Samples
were obtained during the period from April, 1974, through September, 1974,
and from June to September, 1975. Samples were also collected during the
earlier period from \\fhite, urban, middle-class donors residing in metropolitan
Nashville, Tennessee. Donors were asked to complete a brief questionnaire
regarding their exposure to pesticides, their food habits, and. their weight
gain or loss. Samples were kept frozen in polyethylene bags or in glass
bottles until analysis.
DDT and its metabolites were extracted from the samples by slightly
modifying the method recommended by the Primate and Pesticides Effects
Laboratory, Environmental Protection Agency, Research Triangle Park, North
Carolina.33 One milliliter of the milk sample (spiked with 5 ng of aldrin)
was extracted three times with 2.5 ml of acetonitrile in a tissue grinder.
The acetonitrile was added to 25 ml of 2% aqueous sodium sulfate and extracted
3 times with 3 ml of hexane. The hexane extract was concentrated to 0.3 ml
and then fractionated on a Florisil column. The first fraction was eluted
by adding 12 ml of hexane followed by 12 ml of 1% methanol in hexane. The
second fraction was eluted with another addition of 12 ml of 1% methanol in
hexane. Each fraction was concentrated to 500 yl for injection into the gas
chromatograph in a few runs;; because of the small (less than 10 ppb) quanti-
ties of ODD found, the second fraction was generally discarded.
The concentrations of DDT and its metabolites were determined with an
F § M 700 gas chromatograph which was equipped with a Tracer nickel-63
high-temperature electron capture detector. Pesticide separation was
accomplished on a 6' x %" o.d. glass column packed \^ith 1.5% OV-17/1.95%
QF-1 on acid-washed, DMCS treated, 80/100 mesh Chromosorb W. Pesticide
identification was verified on another column packed with 4% SE-30/6% OV-210
on Chromosorb W; mass spectral analysis for absolutely positive identification
was not available. The following conditions were used during pesticide
quantitation: volume of injection, 5ul; column temperature, 200°C; column
nitrogen flow rate, 60 ml/min; detector purge flow rate, 20 ml/min.
Standards were prepared from pesticides received from the Primate and
Pesticide Effects Laboratory using the method suggested by them.33 Working
standards were prepared in hexane and contained a mixture of aldrin and the
isomers of DDE, ODD and DDT.
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One standard used contained 10 ng/ml aldrin, 20 ng/ml op'DDE and pp'DDE,
and 40 ng/ml op'DDD, pp'DDD, op'DDT and pp'DDT. A second standard contained
half the concentration of each of these compounds. Samples with excessively
high pesticide concentrations on the first analysis were diluted quantita-
tively with hexane so that a 5 yl injection would deliver a quantity of each
pesticide that would be less than that contained in the more concentrated
standard. The samples were analyzed by injecting one of the standards and
then running four samples; this was followed by injecting the remaining
standard. Each pesticide concentration was determined by measuring peak
heights and then interpolating between the peak heights of the standards
run on either side of the sample. The limit of detection of this method
was found to be 0.5 ppb aldrin, 1.0 ppb DDE and 2.0 ppb ODD and DDT in the
milk.
Analyses of distilled water samples established that reagents were not
introducing spurious results. Preliminary studies using commercial cow's milk
indicated that the pesticide levels present were below the detection limit
of the method. Eight samples of fresh cow's milk were spiked with the
pesticides of interest and analyzed using the same method as for human milk
samples to check the recovery and precision of the method. Table 2 summarizes
the results. The low recoveries found for DDD were expected since only the
first fraction eluted from the Florisil column was analyzed and the DDD is
known to be split between fraction one and two.3 3 Only fraction one was
analyzed when human milk samples were run. Since the DDD recovered on
preliminary runs was less than 10 ppb, it was decided that the extra time
and effort required to collect and analyzed the second fraction would not be
worthwhile. We confirmed our previous result26 that storage of samples in
polyethylene bags for periods of up to three months does not introduce
spurious peaks in the GLC tracings.
Multiple runs of three human milk samples were made to check the reprodu-
cibility of the method. Table 3 indicates a reproducibility of about ±101 for
analyses over a four-month period, during which all the analyses of the study
were carried out.
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TABLE 2 PESTICIDE RECOVERY STUDIES
Mean amt. Standard
recovered deviation
(%) (8 samples) of
recovery (%)
Aldrin
op1 DDE
pp' DDE
op' ODD
pp' DDD
op1 DDT
pp' DDT
80
96
92
61
28
91
96
± 8
7
6
12
15
5
6
One ml cow's milk-samples were spiked with the pesticides and the samples
were analyzed in the same manner as the human milk samples.
TABLE 3 PESTICIDE RECOVERY REPRODUCIBILITY
Sample
number
5
7
10
Number
analyses
5
5
5
OP' DDE
(ppb)
6 ± 1
8+1
3 ± 0.5
PP' DDE
(ppb)
301 ± 22
1399 ± 188
78 ± 8
OP' DDT
(ppb)
7 ± 1
22 ± 5
3 ± 0.5
PP' DDT
(ppb)
63 ± 13
324 ± 27
36 ± 2
Total DDT*
(ppb)
411 ± 36
1907 ± 230
130 ± 11
*Total DDT is 1.11 x the concentration of both isomers of DDE plus the
concentration of both isomers of DDT. Figures are means ± standard
deviations.
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SECTION V
RESULTS AND DISCUSSION
A total of 38 samples from black donors in rural poverty areas and 14
samples from white, urban, middle-class donors in Nashville in the summer
of 1974 were analyzed. The mean total DDT concentration in the samples
from blacks was 447 ppb with a standard deviation of 465 ppb. The range
of concentrations was from 59 to 1900 ppb (Figure 1). The samples from
the Nashvillians contained a mean total DDT concentration of 75 ppb with
a standard deviation of 40 ppb. The range of concentrations was from 15
to 133 ppb (Figure 2). We had found a mean level of about 170 ppb with
this population 3 years ago.25 An additional 7 samples from the black
donors taken in the summer of 1975 contained a mean total DDT concentration
of 323 ppb \\fith a standard deviation of 176 ppb (range 185-721 ppb).
The mean of the DDT concentrations found in the poverty areas in
1974 was compared with that of the samples from the Nashville area; it was
found that the probability that the means differ due to chance was
8 x 10~5. In all probability, most of this difference can be ascribed
to the greater pesticide exposure of the blacks. However, other factors
may be involved as well. Davies et al.31* and Hoffman et al.35 found
significantly higher DDT concentrations in the adipose tissue of blacks
than in that of whites. The results were obtained by analyzing specimens
of adipose tissue from persons accidentally or violently killed. Although
there were not as many samples from blacks analyzed, the samples from
blacks contained over 50 percent more DDT than the samples from whites.
In our study, all the samples from the rural poverty areas were from
black donors, while those samples from the Nashville area were from white
middle-class donors. A racial comparison of DDT concentrations in human
milk could be made if samples were analysed from middle-class blacks in
Nashville and whites of the same socioeconomic group as the blacks from the
poverty areas; we did not have access to such samples. The probability
that the difference in the means of the tivo sets of samples taken from the
black donors in 1974 and 1975 is due to chance is 0.11. Actually, one
would expect these people to exhibit DDT levels decreasing with time due
to recent reductions in exposure to DDT, and this is in fact observed.
There appeared to be no correlation (correlation coefficient -0.2)
between the age of the black donor and the DDT concentration. It was not
possible to correlate the DDT concentrations with other variables such as
diet, age of child home pesticide use, or distance of residence from
farming fields because many of the questionnaires were not filled out
completely.
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FIGURE 1. FREQUENCY DISTRIBUTION OF TOTAL DDT LEVELS
IN THE MILK OF RURAL BLACK WOMEN (UNSHADED)
AND URBAN WHITE WOMEN (SHADED), 1974
15r-
S!
o
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Total DDT Concentration, ppb
12.
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2. Siyali DS: Polychorinated biphenyls, hexachlorobenzene, and other organ -
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14
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non-target species. Executive Office of the President, Office of Science
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31. Report of the Secretary's Commission on Pesticides and their Relationship^
to Environmental Health-Part I and II, U. S. Department of Health, Educa-7
tion, and Welfare, 1969.
32. Ruckelshaus WD: Order Banning General Use of DDT. Washington, D. C.,
Environmental Protection Agency, June "14, 1972.
33. Thompson JF: Analysis of Pesticide Residues in Human and Environmental
Samples. Perrine Primate and Pesticides Effects^ Laboratory, Environmental
Protection Agency, 1972.
34. Davies JE, Edmundson WF, Schneider NJ, Cassady JC: Problems of prevalence
of pesticide residues in humans, in Davies JE and Edmundson WF (eds):
Epidemiology of DDT. Mount Kisco, N.Y., Future Publishing Co., 1972.
35. Hoffman WS, Adler H, Fishbein WI, Bauer FC: Relation of pesticide con-
centrations in fat to pathological changes in tissues. Arch Environ
Health 15: 758-765, 1967.
15
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TECHNICAL REPORT DATA
'Please read Instructions on the reverse before completing)
1 REPORT NO.
EPA-600/1-76-032
3. RECIPIENT'S ACCESSI ON-NO.
4 TITLE AND SUBTITLE
DDT LEVELS IN MILK OF RURAL INDIGENT BLACKS
6. PERFORMING ORGANIZATION CODE
5. REPORT DATE
September 1976
7 AUTHOR(S)
Bennie T. Woodard, Bruce B. Ferguson and David J.
Wilson
8. PERFORMING ORGANIZATION REPORT NO.
9 PERFORMING ORGANIZATION NAMf AND ADDRESS
Meharry Medical College
MCH/FP Center
Box 69A
Nashville, TN 37208
10. PROGRAM ELEMENT NO.
1EA615
11. CONTRACT/GRANT NO.
R802500
12. SPONSORING AGENCY NAME AND ADDRESS
Health Effects Research Laboratory
Office of Research and Development
U.S.Environmental Protection Agency
Research Triangle Park, N.C. 27711
13. TYPE OF REPORT AND PERIOD COVERED
14. SPONSORING AGENCY CODE
EPA-ORD
15 SUPPLEMENTARY NOTES
16. ABSTRACT
Human milk samples from low-income blacks residing in rural Mississippi and
Arkansas and middle-class whites residing in metrolopitan Nashville, Tennessee,
were analyzed for DDT and its metabolites. The mean total DDT (DDE + DDT) whole
milk concentration of 38 samples from indigent blacks between April and September,
1974 was 447 ppb (range 59 to 1900 ppb) and the mean of the 14 samples from
Nashvillians during the same period was 75 ppb (range 15 to 133 ppb). Seven
samples from the black population in June-September 1975 contained a mean total
DDT of 323 ppb (range 185-721 ppb).
This statistically significant difference in the DDT concentrations in the
black and white populations indicates that the indigent blacks are still highly
contaminated with pesticides even though the general use of DDT has been banned.
Due to the limited amount of information from the donors, no correlation could
be made between the DDT concentration and any factors other than race or socioeconomic
group.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS c. COSATI Field/Group
DDT
Females
Milk
Specimens (statistics)
Population (statistics)
is. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19 SECURITY CLASS (This Report)
UNCLASSIFIED
20 SECURITY CLASS (This page)
UNCLASSIFIED
06, F, T
21 NO OF PAGES
21
22. PRICE
EPA Form 2220-1 (9-73)
16
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