PROCEEDINGS
CATALYST RESEARCH PROGRAM
PLATINUM RESEARCH REVIEW
CONFERENCE
Quail Roost Conference Center
Rougemont, North Carolina
December 3-5,1975
U.S. Environmental Protection Agency
Office of Research and Development
Health Effects Research Laboratory
Catalyst Research Program
Research Triangle Park, North Carolina 27711
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PROCEEDINGS
CATALYST RESEARCH PROGRAM
PLATINUM RESEARCH REVIEW CONFERENCE
Quail Roost Conference Center
Rougemont, North Carolina
December 3-5, 1975
Sponsored by:
U.S. Environmental Protection Agency
Office of Research and Development
Health Effects Research Laboratory
Catalyst Research Program
Research Triangle Park, North Carolina 27711
April 1976
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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. Mention of trade names or commercial products does
not constitute endorsement or recommendation for use.
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CONTENTS
I. Introduction
A. Background 1
B. Objectives 4
II. Conference Presentations
A. Listing of Presentations 6
B. Introduction to Conference 10
C. Highlights of Presentations
1. Baseline Studies 10
2. Measurement Technology and
Emission Characterization 13
3. Methylation Chemistry 20
4. Toxicology and Health Effects 20
III. Workshop Summaries and Recommendations
A. State-of-the-Art and Recommendations
of the Technology Workshop 26
B. State-of-the-Art and Recommendations
of the Health Sciences Workshop 28
IV. List of Participants 33
V. Appendices
I. Abstracts of Presentations
II. General Information for Investigators
111
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I. INTRODUCTION
A. Background
The Catalyst Research Program (CRP) was initiated in FY 1974 by the
Environmental Protection Agency (EPA) Administrator Russell Train concurrent
with the Agency's decision (Nov. 1973) to permit the use of the noble metal
oxidation catalyst as an emission control device on 1975 model year passenger
vehicles. Some research had been initiated prior to this specific program
when EPA first learned of the automotive industry's intent to control vehicle
emissions via the oxidation catalyst. Results of this early research
suggested that, though emissions of hydrocarbons, carbon monoxide, and
certain organics would be dramatically lowered by the catalytic converter,
sulfate emissions would increase, and slight emissions of platinum, palladium,.
and alumina might also be expected.
The possibility of emissions of platinum and/or palladium was of
concern because this would likely represent the introduction of a "new"
environmental contaminant. Though both are found in minute quantities in
the earth's crust, known exposure to these elements had been solely
occupational. It was expected that the active components of the catalyst,
platinum (Pt) and palladium (Pd), would be emitted during vehicle aging as
attrition products. Laboratory studies had measured small amounts of
platinum from automotive catalysts. Early studies with prototype catalyst
vehicles indicated measurable platinum quantities were emitted. Since
platinum is the predominant component of noble metal catalysts, EPA's
research program focused on that element, though palladium was also
studied to a lesser degree in some projects.
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The Catalyst Research Program's approach to assessment of health
effects of both sulfate and noble metals emissions from the catalytic
converter can be outlined as follows:
1. identify emissions
2. determine emission factors
3. estimate human exposure
4. assess public health impact
5. evaluate control options
6. provide timely reports to provide basis for Agency
regulatory decisions.
The study of sulfate emissions from catalytic converter-equipped
vehicles was basically one of assessing the impact of an incremental
contribution from these vehicles to an already existing atmospheric
sulfate burden. Research on characterization, formation, transport,
ambient measurement, exposure and health effects of sulfate had been
ongoing within EPA's Office of Research and Development for some time.
The challenge was to identify effects of sulfuric acid as opposed to
sulfate as a class. Study of the health effects of the noble metals,
however, presented an entirely different challenge. Very little data on
the toxicology of noble metals existed in the literature, and limits of
detection through state-of-the-art measurement technology was not suffi-
ciently low to quantify the expected (but undefined) levels in air,
water, soil, and biological tissue.
The Catalyst Research Program's objective as it pertains to the
noble metals then, was to develop an information base on noble metals,
particularly platinum, in many areas. Time was at a premium, as the
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introduction of catalytic converters was scheduled for the 1975 model
year, to be placed on sale in the fall of 1974. In California, all
American made 1975 model year automobiles would be equipped with noble
metal catalytic converters; in the other 49 states, some 60% would be so
equipped. The number of catalyst-equipped vehicles was expected to
increase rapidly over the next ten years, replacing the pre-1975 catalyst
vehicle population at a rate of about ten percent per year.
The areas of study for the Catalyst Research Program's (CRP). platinum
research effort was set up as shown in Table I.
Table I
Areas of Study, Noble Metal Health Effects
EPA Catalyst Research Program
Emission Characterization (Exhaust) Health Effects Assessment
Measurement Methodology Development Carcinogenicity
Mutagenicity
exhaust Cytogenicity
ambient air, water, soil Inhalation Toxicology
animal & human tissue, Sensitization (allergic
feces, urine response)
standard reference material Irritation
development Immunology
Bioenvironmental Impact Body Burden Tissue Analysis
Methylation Chemistry Epidemiology
Population Exposure Studies
These areas were identified as requiring further research after
receipt of a literature survey conducted by an EPA contractor in FY
1974. Most of the 20-odd research tasks in the CRP associated with
noble metals, particularly platinum, were initiated in FY 1975.
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Work on most of the tasks in the CRP's platinum research effort had
been under way for almost a year at the end of fiscal year 1975. At that
time it became apparent that the data being reported by researchers
should be discussed at an appropriate forum for informative exchange and
discussion. The decision was made early that only active researchers
performing work under EPA contracts and grants, their project officers,
CRP staff, and a very few non-EPA active researchers would be invited to
participate. It was decided that the meeting would not be public, as
data presented would eventually be published as EPA reports upon completion
of the work. In this way, informal exchange of information regarding
work in progress would be encouraged and discussion would not be inhibited
by the possibility of "outsiders" drawing premature conclusions from
preliminary research results reported.
B. Objectives
The conference presentations and the workshops were divided into
two broad categories: (1) analytical and measurement technology, and
(2) health sciences. The main objectives of the conference were to bring
together active researchers in the platinum area to:
1. exchange knowledge
2. identify areas for further study and possible changes in
emphasis or direction of current study
3. recommend priorities
The two-day conference program combined plenary sessions with intro-
ductory and keynote presentations, presentations of results of data for
platinum research tasks included in the CRP, workshop discussions, presenta-
tion of workshop reports, and recommendations by all participants.
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The questions to which workshop discussions were addressed were:
1. Based upon the current state-of-the-art, what conclusions
can now be drawn?
2. What research gaps exist which must be addressed to assess
more completely the environmental consequences of platinum?
3. What is the priority in filling these gap areas?
4. If we (EPA) fill these research gap areas, what additional
conclusions can be drawn?
II. Conference Presentations
During the first day of the conference and continuing through the
early part of the second day's session, a total of 25 presentations on
work completed or in progress were made. A following section, "Highlights
of Presentations," provides a brief description of each project upon
which reports were made.
The twenty-five conference presentations were organized as follows:
Baseline Studies - 3 presentations
Measurement Technology & Emission Characterization - 10 presentations
Methylation Chemistry - 1 presentation
Toxicology & Health Effects - 11 presentations
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A. Listing of Presentations
1. Welcome and Introductory Remarks
Mr. John B. Moran
Environmental Protection Agency
2. "Noble Metals: A lexicological Appraisal of Potential New
Environmental Contaminants"
Dr. Paul E. Brubaker
Environmental Protection Agency
3. *"Review of Literature, Health Effects of Noble Metals, and
Pre-Catalyst Metals and Pre-Catalyst Baseline Population Studies
for Determination of Human Body Burden of Platinum and Palladium"
Dr. Donald Johnson
Southwest Research Institute
Contract # 68-02-1274
Final Report: Baseline Levels of Platinum and Palladium in
Human Tissue
EPA-600/1-76-019
4. "Overview of the Los Angeles Catalyst Study"
Mr. Charles Rodes
Environmental Protection Agency
5. "Determination of Human Body Burden Baseline Data of Platinum
Through Autopsy Tissue Analysis"
Ms. Vandy Duffield
Environmental Protection Agency
6. "Measurement of Platinum from Catalyst-Equipped Vehicles,
Combustion and Attrition Products"
Mr. John Sigsby
Environmental Protection Agency
7. "Toxicological Studies of Auto Exhaust Catalyst Components"
Dr. Jerry Stara
Environmental Protection Agency
8. "The Use of Chemical lonization Mass Spectrometry for the
Analysis of Platinum and Palladium"
Dr. Terence Risby
Pennsylvania State University
Grant # R-803651
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9. * "Chemical Transport of Platinum from Automotive
Catalysts -- A Laboratory Study"
Dr. John Hawley
New York State Department of Environmental Conservation
Former Grant # R-801486
Final Report: The Chemical Transport of
Platinum from Automotive Catalysts
10. "EPA's Efforts to Determine the Platinum Content of Ambient Air"
Dr. Richard Thompson
Environmental Protection Agency
11. "Platinum Metals in Air Particulates Near a Catalytic Converter
Test Site as Measured by Isotope Dilution SSMS"
Dr. Joel Carter
Oak Ridge National Laboratories
Interagency Agreement # EPA-IA6-D5-0466
12. "Measurement of Platinum and Palladium in Ambient Air, Soil, and Water'
"Measurement of Platinum and Palladium in Biological Tissues"
Dr. Donald Johnson/Mr. John Prevost
Southwest Research Institute
13. "Platinum Analyses in Animal Tissues and Fluids"
Dr. Andre LeRoy
National Institutes of Health
14. "Spontaneous Deposition Radiochemical Separation for the
Determination of Platinum, Palladium, and Gold in Biological
and Environmental Materials"
Dr. Donald A. Becker
National Bureau of Standards
15. * "Measurement of Platinum in Biological Tissues, Urine,
and Feces:
Dr. Anna Yoakum
Stewart Laboratories
Contract # 68-02-0663
Final Report: Trace Metal Analysis of Maternal/Fetal Tissue Sets
House Dust Specimen, and Human Scalp Hair
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16. * "Methylcobalamin: Methylation of Platinum and Demethylation
With Lead"
Dr. Robert Taylor
Lawrence Livermore Laboratories
Interagency Agreement # EPA-IAG-D4-0439
Final Report: Comparative Methylation Chemistry
of Platinum, Palladium, Lead, and
Manganese" EPA-600/1-76-016
17. "Toxicity of Platinum (IV) Salts for Cells of Pulmonary Origin"
Dr. Mike Waters
Environmental Protection Agency
18. "Compare Pulmonary Carcinogenesis of Platinum Group
Metal Compounds and Lead Compounds in Association
With Polynuclear Aromatics Using In Vitro Hamster System"
Dr. Philip Kane
State University of New York - Stoneybrook
Contract # 68-02-1299
19. "Studies of No-Effect Level of Platinum and Palladium Using
Mouse Pulmonary Infectivity Model"
Dr. Richard Ehrlich
IIT Research Institute
Contract # 68-02-1273
20. "Compare Relative Toxicities of Platinum Compounds
Against Lead Compounds Using In Vitro Macrophage System"
Dr. Donald Gardner, EPA for
Dr. Eula Bingham
University of Cincinnati
Contract # 68-02-1286
21. "Allergic Induction in Experimental Animals Using
PtS04 and PdS04"
Dr. James Taubler
St. Vincent College
Grant # R-803036
22. "The Effects of Platinum Sulfate on Leucocyte
and Platelet Metabolism and on Immunologic Responses
in the Rabbit"
Dr. Kenneth Lunan
Stanford Research Institute
Contract # 68-02-1300
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23. "Cytogenetic Analyses of the Effects of Platinum"
Dr. Ann Mitchell
Stanford Research Institute
Contract # 68-02-1300
24. *"Toxicities of Compounds of Platinum and Palladium as Measured
by Biochemical Parameters"
Dr. David Holbrook
University of North Carolina
Final Report: Assessment of Toxicity of Automotive
Metallic Emissions
EPA-600/1-76-010A Volume I: Assessment of Fuel
Additives Emissions Toxicity via Selected Assays
of Nucleic Acid and Protein Synthesis
(Contract # 68-02-1205)
EPA-600/1-76-010B Volume II: Relative Toxicities
of Automotive Metallic Emissions Against Lead
Compounds Using Biochemical Parameters
(Contract # 68-02-1701)
25. "Evaluation of the Mutagenic Potentials of Platinum Compounds"
Dr. Shabeg Sandhu
North Carolina Central University
Grant # R-803299
26. "Development of Sensitive Biochemical and Behavioral Indicators
o of Trace Substance Exposure"
Dr. Edward Massaro
State University of New York - Buffalo
Contract # 68-02-1768
*These final reports are available and can be obtained from the project
officers and grantees listed. Those reports that have been assigned an
EPA publication number can also be obtained from the National Technical
Information Service, Springfield, Virginia 22161.
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B. Introduction to Conference
Welcoming remarks and an overview of the conference objectives were
presented by Mr. John Moran, Director of the Catalyst Research Program.
Dr. Paul Brubaker, of EPA and former project officer for some CRP platinum
contracts, discussed the potential toxicological hazards of platinum and
palladium in relation to their use in catalytic converters.
C. Highlights of Presentations
1. Baseline Studies
(a) The Southwest Research Institute (SwRI) study of platinum
and palladium was reported by Dr. Donald Johnson. The first phase of
the study was a literature survey performed in early 1974 covering
sources, uses, production, consumption, reported medical cases, and
toxicology of platinum and palladium. The literature survey concluded
that "no data existed by which an estimate can be made of transfer of
platinum and palladium to the environment. Investigations show that
only salts of platinum present human health hazards. Industrial exposure
to these is limited to the mining and refining of platinum ores and the
preparation of catalysts for the chemical and petroleum refining industries."
The second phase of SwRI's study covered several areas:
(1) collection and analysis of biological samples (blood, urine,
hair, and feces) and environmental samples (air, soil, and
water) from populations in southern California -- one in an
urban area near a major freeway, the other in a non-urban
location. The urban population studied was near the Los
Angeles Catalyst Study site which allowed for corroborative
pollutant and meteorology data comparison. The site was discussed
in Mr. Rodes1 presentation.
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(2) Collection and analysis of biological samples and environmental
samples from three locations -- one at a mining and processing
facility, and two at platinum/palladium refineries.
(3) Limited autospy samples were collected from the area near the
area of study discussed in (1) and near the mining area described
in (2).
Preliminary conclusions drawn from the results of the Pt/Pd baseline
studies were reported as follows:
• Baseline levels of Pt and Pd in the environment
(air, soil, water) and in urban populations are extremely
low. Levels in nearly all samples were below the detection
limits of the analytical methods (which, however, were
not defined).
t Examination of ambient air, and blood, urine, and feces
from miners for levels of Pt and Pd did not provide
definitive data on the relationship of ambient air to
body burden, as air levels were nearly always below
detectable levels, which, however, were not defined.
• Data from Pt and Pd refineries indicate that measurable
levels of Pt (0.23.- 2.6 yg/liter) and Pd (0.21 - 6.3
ug/liter) are found in the urine of workers exposed to
o o
air containing 0.02 - 0.26 yg/m Pt and .003 - 0.36 yg/m
of Pd. Measurable levels were not found in the blood of
these workers.
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t Data from the refineries indicate that it is likely that
measurable levels of Pt and Pd could be seen in the
general population when ambient levels of these two
metals approach 0.1 ug/m . The metals would appear
initially in urine rather than blood.
t Additional data are needed on the distribution of Pt and
Pd in human tissue other than blood and urine following
exposure to air containing the 2 metals at the 0.01 - 0.1
ug/rn .levels. Additional data to define the,relationships
between air levels and Pt and Pd and tissue levels in the
general population are needed, but development of data
should be;deferred until there is. more-information available
on quantities and types of the two me;taIs. emitted from
the catalytic mufflers.
(b) The second baseline presentation, made by Ms. Duffield,
reported data resulting from^analysis,by.;Stewart Laboratories of autopsy
samples collec,tedvby EPA in southern California* the same general area
where two of the populations .studied^in the Southwest Research Institute
work were located.. Results of analysis, for Pt ,in 97 autopsy sets were
presented. Analysis .was performed by a specially developed ,emission
spectrochemical method. .Almost half.,(46%) of the .individuals studied
were1 found to have detectable'-amounts of platinum in ,one ,or.more tissue
samples. -Levels of platinum in .13 of-21 tissue types were detected.
The range of Pt concentrations was 0.003 to 1.46. vg/gm,(wet..tissue); the
mean value of detected Pt was 0.16 ug/gm, and the median value of detected
Pt was 0.067 ug/gm. Surprisingly high values were found in subcutaneous
fat, not previously considered to be a target site.
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(c) A brief report of set-up and operations at the Los Angeles
Catalyst Study (LACS) site was presented by Charles Rodes. This monitoring
site, which consists of four instrumented stations located in a perpendicular
line across a major freeway in the Los Angeles area, is an important
support operation to the CRP platinum research effort. It was at this
location that a large particulate sample was collected to determine
baseline ambient Pt. Also, one of the southern California populations
studied in the SwRI human body burden baseline study was located near
downwind of the LACS site, thus presenting corroborative ambient pollutant
and meteorological data for analysis of the SwRI study results. If at
some future time it is determined that automotive emissions of Pt are at
a high enough level to warrant concern for inhalation exposure of this
general population, this site would be an important part of any monitoring
effort initiated by EPA.
2. Measurement Technology and Emission Characterization
(a) A brief summary of EPA/ORD's attempts to identify platinum
in exhaust particulate emitted by catalyst-equipped vehicles was presented
by Mr. John Sigsby. Some 20 cars were included in both inhouse and
extramural emissions characterization studies. No platinum was ever
detected in particulate collected on filters, so that if there is any
platinum in the size range of airborne exhaust particulate, it is below
current detection limits. These detection limits equate to < 10~
gin/mile in the extramural activities. On the basis of these measurements,
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it is EPA's conclusion that there is at present no risk to public health
presented from exposure to respirable range platinum in catalyst-equipped
vehicle exhaust particulate. However, no measurements were made with
vehicles other than those operating properly and with unleaded fuel.
Platinum has been identified in the larger size range of exhaust
particulate (that portion of the exhaust which would normally deposit
upon the roadway or near the roadway). Analysis was performed on tube
sweepings collected from the bottom of dilution tunnels in which the
exhaust is diluted prior to collection of samples for both particulate
and gaseous analysis. The collections were made after many runs, often
with a variety of properly operating catalyst vehicles. One sample was
split and analyzed by a variety of techniques, others were only analyzed
by atomic absorption. The material collected was extremely heterogeneous
consisting of a wide range of sizes including a few whole catalyst
pellets. The range of Pt content found in various samples and fractions
ranged from 0.034 to 635 yg/gm of collected material. The higher values
were related to whole pellets and pellet fragments and corresponds with
the initial catalyst loadings. Such pellets and fragments are usually
only found on initial runs on fresh unused catalysts. Emission spectroscopy
for chemical analysis (ESCA) results showed that some of the platinum
appeared to be in the +4 oxidation state but no quantisation was possible.
A definitive characterization has not yet been performed. There is
question as to whether precise identification is necessary or possible
due to the small amounts emitted and to the catalyst matrix with which
it is associated.
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(b) Dr. Stara described work performed by EPA's Cincinnati
group wherein animals were exposed to both catalytic and non-catalytic
automobile exhaust. In an early exposure experiment with a prototype
catalyst, platinum was found in chamber air near the detection limits of
atomic absorption (AA) only once; in later experiments with production
catalysts, platinum was found once in a filter placed near the vehicle's
tailpipe, again near the limits of detection for AA. Four samples of
large particulate collected in the dilution tunnel were analyzed for
platinum by flame atomic absorption at EPA's North Carolina Environmental
Research Center. Though there were matrix difficulties, valid results
were obtained using standard additives. Platinum was found in the
samples in concentrations ranging from 60 to 145 yg of Pt per gram of
the samples. No attempt was made to identify the platinum compound due
to small amount of samples available for analysis.
(c) Dr. Risby of Pennsylvania State University presented a
method for analysis of volatile mixtures. He described his preparation
of volatile chelates of platinum and palladium. Using a chemical ionization
mass spectrometric method, spectra were established for the noble metal
chelates synthesized. Limits of detection for the compounds are in the
picogram range.
(d) Dr. John Hawley of the State of New York Department of
Environmental Conservation described results of parallel bench laboratory
and engine stand tests of chemical transport of platinum from noble
metals oxidation automotive catalysts. Scaled to a 60 mph cruising
automobile, particulate Pt was collected during some laboratory experiments.
Engine stand tests also showed microgram per mile platinum transport
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under certain conditions. Tests conducted with nitrogen and oxygen as
the carrier gas showed that platinum, in these experiments, was transported
chemically rather than mechanically. Analysis was performed by x-ray
fluorescence.
(e) Dr. Thompson discussed EPA's efforts to determine the
platinum content of ambient air. In 1969 massive samples of suspended
particulate matter collected from ambient air were analyzed for all
elements having stable nuclides, except the noble gases. Results of
analysis for platinum by X-Ray Fluorescence (XRF), Optical Emissions
Spectroscopy (OES), Atomic Absorption Spectrometry (AAS), and Neutron
Activation Analysis (NAA) were negative. In 1975, the recently developed
isotope dilution mass spectrometry technique showed platinum existing at
—fi ^
a level of 2.5 x 10" ug/m (based on an assurned air volume obtained by
considering the particulate to have come from air at, an average TSP
loading).
.(f) Dr. Opel.Carter described the method by which isotope
dilution spark-source mass specjtrpmetry was .employed at Oak Ridge National
Laboratory for analysis for platinum of.the above-mentioned massive
suspended particulate samples.. To obtain sufficient,sensitivity for
obtaining positive values for platinum, it was necessary to dissolve a
large portion, of the,ashed sample for the purpose of equilibrating
1Qfi 10? Qft
stable.isotopes of Pt, u Pt, and_ Ru. .Platinum as a metal prec.ipi.tate,
and gold as a carrier were then added. Gold and the platinum..group were.
concentrated into .the gold precipitate used ,in the.isotope .dilution;
spark-source mass spectrometry .measurements. As mentioned earlier,
platinum levels were reported at 2.5 picograms per cubic meter of.air.
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(g) Dr. Don Johnson of SwRI presented results of analysis of
air, soil, and water samples collected at the various sites included in
the baseline study mentioned above (see "Baseline Studies"). Using a
method involving digestion extraction and preconcentration followed by
atomic absorption, "less-than" values were reported for all samples
except air samples collected inside mining and refining facilities. Air
concentration levels of platinum ranged from 0.021 - 0.259 ug/m for
platinum refinery locations (mean of 0.159 ug/m ); only one positive
value was reported in the mining facility (0.377 ug/m Pt). For a more
detailed description of analytical methods employed and limits of detection
applicable for the various types of samples, see final report.*
Measurement of platinum in biological samples was reported on by
four conference participants. Dr. Johnson reported that the method for
biological samples is similar to that previously discussed for platinum
analysis of environmental samples. No detectable levels of platinum
were found in individual samples of blood, urine, hair, and feces
collected from the California residents (both urban and non-urban)
participating in the study. Analysis of a composite of all blood from
all age groups and both sexes gave a value of 0.049 ug/100 ml for the
group near the Los Angeles freeway and 0.180 ug/100 ml for the high
desert non-urban group.
No detectable levels of Pt were found in blood, urine, hair, and
feces collected from miners at the mining site. At the Pt refinery
*Directions for obtaining copy of final report on page 9.
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location, positive values were detected for platinum in urine in a range
of .23 - 2.5 yg/liter, where minimum detectable Pt concentration was
0.10 yg/1.
Autopsy samples were collected from cases, again, near both the Los
Angeles site and near the mining area. Only "less than" values were
reported from the two groups numbering 10 and 9 cases, respectively,
with the exception of 3 tissue samples from 3 of the nine mining area
cases. Analysis for platinum in these 3 cases gave Pt values for fat,
lung, and muscle samples at 4.5, 3.7, and 25.0 ppb, respectively.
(h) Work in the area of platinum analysis of animal tissue
and fluids performed at the National Institutes of Health was described
*
by Dr. Andre LeRoy. An adaptation of the flameles atomic absorption
spectrophotometry method has given quantitative determination of platinum
in biological samples down to about 30 parts per billion. The NIH
analytical method is similar to the SwRI method for Pt analysis by
atomic absorption, but differs in the digestive and preconcentration
procedures. In the NIH method, tissues are digested first by nitric
acid, then by a mixture of perchloric and nitric acid. The digest is
taken up in HC1, aliquots of which are analyzed by comparison with a
working standard. Aliquots of blood plasma or urine are analyzed directly
for samples containing a minimum Pt content of 2 ng/30ul. Detection
limits are reported to be approximately 30 ppb.
An important feature of the method used is the introduction of a
temperature ramp generator developed and built at NIH which replaces the
step-wise temperature change in the graphite furnace with a ramp. Its
use results in a time interval between absorption peaks due to "smoke"
and molecular absorption from the sample combustion and the atomic
absorption by the ground state platinum atoms.
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(i) Mr. Donald Becker of National Bureau of Standards described
the application of neutron activation analysis coupled with spontaneous
deposition radiochemical separation to determination of platinum and
other noble metals in biological and environmental materials. Sample
preparation involves lypholization and wet-ashing with a mixture of
nitric and perchloric acids on a hot plate, dissolution of the perchlorate
residue in hydrochloric and nitric acids, and addition of "holdback"
carriers prior to the addition of silver powder to effect deposition.
The silver powder is then counted on a germanium gamma-ray detection
system. Results of an analysis of bovine liver and orchard leaves gave
platinum values of <1.6 ng/g and <9 ng/g, respectively. Analysis of
sample splits with Dr. LeRoy of NIH for biological tissues from animals
which had ingested platinum compounds indicated reasonably good agreement
between the two methods.
(j) Stewart Laboratories' literature search for available
methods for analysis of platinum conducted in 1974 and subsequent
development of a method specifically for determination of trace quantities
of platinum, lead, and manganese in biological tissues was presented by
Dr. Anna Yoakum. The sample preparation for the method involves lypholization
and wet-ashing prior to spectrochemical analysis. Samples and standards
are packed into carbon electrodes and analyzed by a total energy burn in
a controlled argon-oxygen atmosphere with dc arc excitation.
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3. Methylation Chemistry
Dr. Robert Taylor of Lawrence Livermore Laboratories described
his work on comparative methylation chemistry of platinum, palladium,
and lead. He reported that at least one platinum compound, potassium
hexachloraplatinate, can be methylated via the same Vitamin B^2 reaction
by which mercury is methylated. By labeling both the carbon and the
hydrogen in the methyl group component of methylcobalamin (Vitamin B^
metabolite), Dr. Taylor established that the platinum compound demethylates
the MeB12 to form a methylated,platinum compound.• This methylated
platinum compound, though slightly light-sensitive, appears to be moderately
stable, but has yet to be fully characterized and evaluated for biological
activity. Characterization will be difficult, as the compound cannot be
lypholized without loss of integrity. Dr. Taylor also reported that
platinic sulfate also demethylates MeB,2, but further work is necessary
to establish whether the methyl, group ;is transferred, to the platinic
sulfate.
Ip. addition, various Pd, Pb, and Mn compounds showed ^significantly r
lower levels of demethylatio.n activity, in,the.^presence of MeB,'? when
compared to plat.inic sulfate.
4. Toxico-logy and Health Effects
, (a) Dr. M-ike,Waters of EPA reported results of ire vitro
studies of acute toxicity of platinum (+4) valence compounds uslnig"
rabbit alveolar macrophages and human lung fibroblasts. Though platinum
dioxide exposure elicited no response, platinum tetrachloride exposure
resulted in significant loss of viability for both macropl^age and fibroblast
cells. Thymidine uptake and incorporation (DNA synthesis) was shown to
be 50% inhibited at a concentration of 10 M PtCl.; a concentration of
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approximately 6 x 10 M was required to produce 50% inhibition of uptake
and incorporation of C uridine (RNA synthesis) and leucine (protein
synthesis).
(b) EPA's Health Effects Research Laboratory in Cincinnati
has performed a number of studies on platinum, involving both biological
fate and toxic response to various compounds of platinum following
various routes of exposure. Dr. Jerry Stara reported on the results of
these studies, some highlights of which were as follows:
• Retention, tissue distribution, and excretion of
191
Pt in rats were determined after intravenous, oral,
and inhalation exposure. A substantial portion of platinum
is excreted rather rapidly in either feces or urine.
Deposition sites included liver, adrenal, spleen, kidney,
lung, or bone. Lung clearance after inhalation exposure
has a half-time of about 8 days. Platinum was found to
cross the "placenta! barrier" in small amounts after
dosing of pregnant rats.
• Platinum salts tested did not appear to affect the
mitochondrial function of the cell in various organs.
• Platinum salts tested were 100 to 1000 times less
active than lead in inhibiting ALA-D enzyme activity.
(c) Dr. Kane described the study being conducted by the State
University of New York at Stoneybrook in which the potential role of platinum
and lead compounds as co-carcinogens in respiratory carcinogenesis
-------
22
is being evaluated. The metals are to be evaluated in conjunction with
the known carcinogen benzo(a)pyrene (BaP). Intratracheal and intragastric
instillation in two groups of hamsters are in progress. Since this is
of necessity a long-term study, no conclusions can yet be drawn as to
the tumor producing properties of platinum compounds.
(d) The studies of no-effect level of Pt and Pd for response
of treated mice to a challenge of influenza virus or Streptococcus sp.
were reported by Dr. Ehrlich of IITRI. Results indicate that inhalation
of platinum and palladium oxide aerosols diminished survival time when
challenged with the infectious agent compared with control mice challenged.
Limited experimentation with response to Staphylococcus aureus administered
via intranasal instillation indicated that inhalation of PtO^ aerosol
dimished lung clearance of the infectious agents, presumably by reduction
of function of the alveolar macrophages.
(e) Dr. Gardner presented results of work done by Dr. Eula
Bingham at the University of Cincinnati. The study involved a comparison
and evaluation of cellular response in the lung (especially alveolar
macrophage) to inhalation of platinum and other metal compounds. Inhalation
exposure to platinum salt did not statistically alter the number of lung
cells, while some other metals administred via inhalation did significantly
alter the number of cells, both positively and negatively.
-------
23
(f) Dr. Taubler reported that studies of immediate and delayed
allergic response to platinum compounds in rabbits, guinea pigs and mice
produced essentially no positive response with metals alone. However,
when complexed to a protein (albumin) the toxicity of both metals was
potentiated to a degree that the majority of animals died when a subsection
was exposed to platinum and a positive allergic response was elicited
from palladium.
(g) Drs. Lunan and Mitchell discussed the several studies
using platinic sulfate being performed at Stanford Research Institute
including effects on leucocyte and platelet metabolism, immunologic
response, and cytogenetic effects in the rabbit. Preliminary data
showed the following:
t Guinea pig skin sensitization studies and rabbit skin
irritation studies gave no positive response.
• Eye irritation experiments with rabbits showed the
PtCSQ^ should be regarded as a moderate to severe eye
irritant.
• In vivo Pt(SQ4)2 treatment only moderately affected
leucocyte metabolism. In the leucocytes from the 1- and
3-week exposure studies DNA and RNA synthesis, respec-
tively, were depressed 50%. In the 3-week exposure plus
1-month recovery study phospholipid turnover was markedly
retarded.
• In vitro experiments indicated that 2-hour exposures to
-4
10 M platinic sulfate caused 50% depression of DNA
synthesis in leucocytes.
-------
24
e Numerous alterations of platelet metabolism were observed
in all experiments involving animals exposed to 15 and 20
mg/kg of Pt(SQ4)2.
• Several whole blood values were affected in recovery
experiments.
Cytogenetic analyses are in progress, but no data are yet
available.
(h) Evaluation of the mutagenic potential of several platinum
compounds in progress at North Carolina Central University was discussed
by Dr. Sandhu. Both an in vitro system involving mouse lymphoma cells
and an In vivo study of multi-generation growth of Phaseolus aurealis
are being employed in this study. Work is now in progress, but data are
not yet available.
(i) Dr. David Holbro.ok of the University of North Carolina
summarized the results of his studies on the -toxicity of platinum com-
pounds as measured by biochemical parameters. Studies of both dietary
intake and drinking water administration of platinum salts showed that
high levels of exposure restricted weight gain., Dietary administration
did not show any significant effect on organ weights.
Intraperitoneal injection of platinum salts at high levels
significantly reduced DNA synthesis in several organs. Also, high
levels of platinum salts administered intraperitoneaMy decreased liver
. -Ji * ' • '
detoxification mechanism activity. Dietary administration showed no
consistent effect on this function.
-------
25
(j) Mr. John Prevost and Dr. Donald Johnson of SwRI discussed
the results of that portion of the baseline studies which examined levels
of platinum in blood, urine, hair, and feces of both groups occupationally
exposed to platinum and those exposed only to general ambient levels.
In the studies of occupationally exposed subjects, it was established
that exposure to levels of .02 - 0.2 ug/m or higher can result in
measurable levels of platinum in urine. No platinum was detected in
blood, hair, or feces of either occupationally on non-occupationally
exposed study participants. A composite blood sample from two non-
occupationally exposed groups showed Pt levels of .049 and .180 yg/100
milliliters respectively, as discussed above.
(k) The last presentation was a report on an experimental
area quite different from its predecessors. Dr. Massaro of the State
University of New York at Buffalo discussed his work in the tissue/organ
distribution and effect of platinum on behavior, as well as levels of
neurotransmitters, glycolysis, and rates of DNA, RNA, and protein synthesis.
Though the results of the experiments are so far preliminary, it appears
that Na2PtClg has an effect on open-field behavior which would be charac-
terized as depressant.
-------
26
III. Workshop Summaries and Recommendations
A. State-of-the-Art and Recommendations of the Technology Workshop
Question 1: Based upon the current state-of-the-art, what conclu-
sions can now be drawn?
1. Pt is emitted from motor vehicles equipped with catalysts.
Emission factors are unknown. Predominant size is non-
respirable. Form is particulate. Composition is unknown.
2. Background exposure to Pt in air, water, and soil is very
low, but data are incomplete because levels are at or
below current measurement capabilities.
3. Occupational exposures at fairly low levels may result in
detectable human burdens. :
4. Autopsy samples from the general population evidence Tow
level burden of Pt, but exposure route is unknown.
Living individual human populations do not evidence Pt
burdens at current limits of detection.
5. Current measurement methods are not adequate to establish
exposure or body burden background.
6. Our current knowledge of environmental burdens of Pt and
distributions thereof are not now adequate to assess
exposures.
Question 2: What research gaps exist which must be addressed to
more completely assess the environmental consequences
of platinum?
-------
27
1. Determine emitted Pt compound(s).
2. Develop adequate analytical techniques focusing upon
concentration and sample preparation step. Once com-
pleted, a number of analytical tools are adequate to do
the job.
3. Ascertain Pt concentrations, reactions, and transport in
the ecosystem (air, soil, sediment, water, localized
exposure, tissue, food, plants).
4. Determine Pt emission factors, size distribution and
form.
5. Develop Pt monitoring analytical capabilities.
6. Develop (EPA) mechanism to assure operational quality
control in multi-task programs such as in the Catalyst
Research Program.
7. Must assure analytical methods intercomparability.
Question 3: What is the priority in filling these gap areas?
1. Determine emitted Pt compound(s).
2. Develop adequate analytical techniques focusing upon
concentration and sample preparation step. Once com-
pleted, a number of analytical tools are adequate to do
the job.
3. Develop Pt monitoring analytical capabilities.
4. Develop (EPA) mechanism to assure operational quality
control in multi-task programs such as in this program.
-------
28
5. Assure analytical method intercomparability.
6. Ascertain Pt concentrations, reactions, and transport, in
the ecosystem (air, soil, sediment, water, localized
exposure, tissue, food, plants).
Question 4: If we fill these research gap areas, what additional
conclusions can be drawn?
1. Determine quantitative environmental Pt distribution.
2. Movement of Pt in the ecosystem.
3. Composition of Pt,exposure.
4. Determine exposures to Pt compounds in the-environment
(basis'for assessment.of pubI.ICLIhealth consequences).
5. Can, in the future, assess quantitative changes in
exposures.
f
B. State-of-the-Art and Recommendations, of the; Health Sciences Workshop
Question 1: Based upon the current state-of-the-art, what conclu-
• sions can now be drawn?
From the literature and papers presented,;it is evident
th'at platinum compounds, a-t some concentrations, can have a
biological effect, i.e., biochemical, immunolpgical, host
defense, and physiological. If such effects;are to be seen in
; the population/they wiImprobably be the result of long-term
low levels of exposure. This necessitates the need for further
testing in the areas of carcinogenesis, mutagenesis, immunological,
behavioral, and biological screening (in vitro test).
-------
29
At the present time, the levels of Pt in air, water, and
soil appear to be very low. It will be important to continue
monitoring atmospheric, soil and water levels in the future.
Increases in levels in these media may likely result in some
biological reaction since the present concentration is so low.
There are many areas that need further investigation. We
know nothing about turn-over-rate nor what form the substance
is in the tissue. Nor do we know what form the Pt is when
emitted from the exhaust. This latter information is vitally
important to the environmental toxicologist in performing
experiments.
Question 2: What research gaps exist which must be addressed to
assess more completely the environmental consequences
of platinum?
The health research needs regarding these potential
environmental toxicants is great and to fulfill all goals
would require a great deal of time and money. There are many
"gaps," but a few essential areas which should be investigated
first are listed below:
1. Most relevant to EPA mission are studies which will
investigate the long-term, low concentration effects
resulting from exposure to platinum compounds.
2. Studies should be conducted to determine deposition and
target site of action of these agents (through different
exposure routes).
-------
30
3. Interaction studies of Pt with other agents that are
being emitted from the exhaust (HpSO.).
4. Characterization of emission/attrition products, and of
ambient samples.
5. Correlation of in vitro test with whole animal studies.
6. Greater involvement of EPA ecological team in following
the biotransformation of Pt in food chain and water.
Coupled with this would be controlled feeding studies.
7. Investigation into the effects on sensitive populations.
8. Continued studies on comparative toxicological effects of
selective Pt compounds.
9. Long-term epidemiological studies of the population at
risk, once defined.
Question 3: What is the priority in filling these gap areas?
In order to obtain the maximum amount of useful information
concerning the potential toxicity of these agents, it is
necessary to study simultaneously the areas of epidemiology,
environmental assessment, and the biological effects. The
group's recommendation for toxicological testing is listed as
follows:
1. Develop information on what concentrations exist in man
(non-occupational). Evidence suggests that Pt is in
tissue, but how did it get there (food chain, H,,0?) In
what form is the Pt?
-------
31
2. Because of the time it takes to obtain the information,
long-term, low-level research studies should begin soon
(carcinogenesis, mutagenesis, host-mediated immunity,
etc.)
3. Development of model systems for study of the sensitive
population (neonates, hypersensitive, developing embryo).
This can be the basis for future epidemiological studies.
4. Research on the interaction of Pt with other exhaust
emissions such as H2S04, NO^.
5. Measurement of Pt in food chain (milk samples, etc.).
6. Study of deposition and target site of Pt once it has
entered the body by various routes of exposure.
It was also indicated that there must be a close working
relationship between the various scientific disciplines as
well as between the contractor and the funding agency. Meetings
such as this one bring together individuals with similar
interest and permit very meaningful scientific dialogue.
Question 4: If we fill these research gap areas, what additional
conclusions can be drawn?
If the above-suggested research is performed, we will be
able to obtain useful information upon which to base decisions
as to the need for any regulatory action. We will be able to
evaluate the potential health hazard(s) resulting from any
predicted increase in levels of exposure to platinum or platinum
compounds. Finally, the data generated in these experiments
-------
32
would provide us with scientific models which could be used in
future toxicological screening of new environmental pollutants.
Evaluation could be made more quickly and accurately of possible
adverse effects on public health.
-------
33
Participants
Platinum Research Review Conference
Mr. Donald A. Becker
Room A 309
Bldg. 222
National Bureau of Standards
Washington, D. C. 20234
Dr. Arthur Bierman
Lawrence Livermore Laboratories
Livermore, California 94550
Dr. Paul E. Brubaker
Environmental Protection Agency (formerly Criteria and Special Studies
RD-683 Office, HERL, EPA, RTP, NC)
Washington, D. C. 20460
Dr. Robert Bruce
Health Effects Research Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Mr. Joseph Bumgarner
Environmental Monitoring and Support Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Dr. Joel Carter
Oak Ridge National Laboratories
Box Y, Building 9735
Oak Ridge, Tennessee 37830
Ms. Vandy Duffield
Health Effects Research Laboratory
Environmental Protection Agency
Research Trianlge Park, North Carolina 27711
Dr. Richard Ehrlich
I IT Research Institute
10 W. 35th Street
Chicago, Illinois 60616
Dr. Edward Faeder
Health Effects Research Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Dr. Donald Gardner
Health Effects Research Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
-------
34
Mr. Herman Gibb
Office of Health and Ecological Effects
Environmental Protection Agency
Washington, D. C. 20460
Dr. George Goldstein
Health Effects Research Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Dr. John Hawley
New York State Department of
Environmental Conservation
50 Wolf Road
Albany, New York 12233
Dr. David Hoi brook
Department of Biochemistry
School of Medicine
University of North Carolina
Chapel Hill, North Carolina 27514
Dr. Donald Johnson
Southwest Research Institute
8500 Culebra Road
San Antonio, Texas 78284
Dr. Philip Kane
State University of New York
Department of Pathology
Health Sciences Center
Stoneybrook, New York 11790
*
Dr. Andre LeRoy
Bldg. 13
Room 3W 13
National Institutes of Health
Bethesda, Maryland 20014
Dr. Kenneth Lunan
Stanford Research Institute
333 Ravenswood Avenue
Menlo Park, California 94025
Dr. Edward J. Massaro
State University of New York - Buffalo
Dept. of Biochemistry
School of Medicine
Buffalo, New York 14212
-------
35
Dr. Robert Miller
Health Effects Research Laboratory
Environmental Protection Agency
Cincinnati, Ohio 45268
Dr. Ann Mitchell
Stanford Research Institute
333 Ravenswood Avenue
Menlo Park, California 94025
Dr. Wellington Moore
Health Effects Research Laboratory
Environmental Protection Agency
Cincinnati, Ohio 45268
Mr. John B. Moran
Director, Monitoring Technology Division
Environmental Protection Agency (formerly Director, Catalyst
RD-680 Research Program, HERL, EPA,
Washington, DC 20460 RTP, NC)
Mr. John Prevost
Southwest Research Institute
8500 Culebra Road
San Antonio, Texas 78284
Dr. Terence Risby
Pennsylvania State University
424 Davey Laboratory
University Park, Pennsylvania 16802
Mr. Charles Rodes
Environmental Monitoring and Support Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Dr. Shabeg Sandhu
North Carolina Central University
P.O. Box 19553
Durham, North Carolina 27707
Mr. John Sigsby
Environmental Sciences Research Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Dr. Jerry Stara
Health Effects Research Laboratory
Environmental Protection Agency
Cincinnati, Ohio 45268
-------
36
Ms. Peggy Stewart
Stewart Laboratories, Inc.
5815 Middlebrook Pike
Knoxville, Tennessee 27921
Dr. James Taubler
St. Vincent1s College
Biology Department
Latrobe, Pennsylvania 15651
Dr. Robert Taylor
Lawrence Livermore Laboratories
Livermore, California 94550
Dr. Richard Thompson
Environmental Monitoring and Support Laboratory
Environmental Proteciton Agency
Research Triangle Park, North Carolina 27711
Dr. Michael Waters
Health Effects Research Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Dr. Anna Yoakum
Stewart Laboratories, INc.
5815 Middlebrook Pike
Knoxville, Tennessee 27921
-------
APPENDICES
I. Abstracts of Presentations
II. General Information for Investigators
A. Platinum Chemistry Pertinent to the Catalyst Program
B. Spectrophtometric Verification of PtCU"
-------
APPENDIX I
Abstracts of Presentations
-------
Environmental Health. Perspectives
Vol. 10. pp. 39-56. 1975
Noble Metals: A Toxicological Appraisal of
Potential New Environmental Contaminants
by Paul E. Brubaker,* John B. Moran,* Kenneth Bridbord,* and F. Gordon
Hueter*
The public health benefits expected by reducing known hazardous emissions from mobile
sources should not be compromised by increasing levels of other potentially hazardous
unregulated emissions. Catalytic converters are going to be used to meet the statutory
requirements on carbon monoxide and hydrocarbon emissions from light duty motor
vehicles. Platinum and palladium metals are the catalytic materials to be used in these
emission control devices. Preliminary experimental evidence and analysis of the impact
of these control devices on the future use and demand for platinum indicates that this
mutal may appear at detectable levels in the environment by the end of this decade. At
the present time, platinum and palladium are not present in the public environment and
represent potentially new environmental contaminants as a consequence of use of this
new abatement control technology. There is relatively little information available to
adequately assess the potential health hazards that may be associated with exposure to
these metals and their compounds. Analysis of the environmental problems and concerns
associated with possible new environmental contaminants arc discussed. Limited esti-
mates are made on community exposure by use of a meteorological dispersion model.
Biodegradatiotr potential and attention is also given to the limited toxicological infor-
mation available.
* Special Studies Staff, Office of the Director,
National Environmental Research Center, Research
Triangle Park, North Carolina 27711.
1-A
-------
Review of Literature, Health Effects of Noble Metals
and Pre-Catalyst Baseline Population Studies for
Determination of Human Body Burden of Pt and Pd.
Dr. Donald E. Johnson
A summary of the literature review of the supply and demand
for platinum and palladium will be presented. This review includes
the prediction of the impact of the catalytic muffler on these markets.
Data on the health effects from the two metals will also be summarized.
The justification for a baseline study and for surveys to help understand
the relationship of exposure to the two metals and body burdens will
be presented.
2-A
-------
OVERVIEW OF THE LOS ANGELES CATALYST STUDY
by
Charles E. Rodes
Environmental Monitoring and Support Laboratory
Environmental Monitoring Branch
Research Triangle Park, North Carolina 27711
Summary of Presentation at
Platinum Research Review Conference
Quail Roost Conference Center
December 4,
3-A
-------
The Los Angeles Catalyst Study (LACS) is a field monitoring
program which was designed to determine the impact of catalyst equipped
cars on the ambient air in the vicinity of a Los Angeles freeway.
The principal pollutants of interest in this study are H^SO. and
sulfate aerosols. Since an acceptable monitoring technique is presently
not available for HpSO* aerosol, associated pollutants and parameters
such as Pb, SCL, CO, and meterology are monitored to allow "surrogate"
predictions of the expected ambient HpSO^ concentrations.
The site selected for the LACS is adjacent to the San Diego
Freeway approximately 0.5 miles north of the Wilshire Blvd interchange.
See Figure 1 for the location in Los Angeles and Figures 2 and 3
for the layout of the sites and the location of the instrumentation.
Site selection criteria included: (1) a traffic flow (180-200,000
cars per day), (2) prevailing cross-freeway winds, (3) lew S02 background
levels, and (4) site accessibility. Sampling sites were established
on a perpindiculer line to the freeway to allow the contributions
from the freeway to be determined by simultaneous up-wind and down-
wind measurements. A system has been recently installed to provide
information on the traffic speed and count by lane.
The LACS has been on-line continuously since June, 1974, to
obtain data before and after the introduction of the catalyst cars.
An annual report of the data collected during the first year was
4-A
-------
prepared in July, 1975. Some typical data for the May, 1975, are
shown in Tables 1 through 9. Tables 1 through 5 show data from the
1500-1900 hour interval (evening rush-hour) when there is not only
peak traffic flow, but also consistent perpindicular cross-freeway
winds. Tables 6 through 9 are hourly data reduced to 24-hour averages,
Figure 4 is a summary of the prevailing wind directions and wind
speeds for May, 1975. Table 10 is a sample of the hourly traffic
count data for November 10, 11, and 12, 1975.
The Pb data in Table 2 shows a monthly average contribution
o
by the freeway of 7.40 vg/m (Site C minus Site A). This can be
compared to a yearly average contribution of 5.83 pg/m .
There is no on-going program to measure Pt in the LACS program,
because of analytical sensitivity limitations. A single large dust
sample was collected from the air conditioning system of a building
near the LACS site during the pre-catalyst period. This sample was
analyzed for Pt by the Analytical Chemistry Branch (ACB) of EMSL/RTP
and a subsequent report is available. There are only two projects
on-site from which the Pt data could be obtained. One is through
analyses of soil and vegatation samples at various distances from
the freeway. The other would be to composite into one large sample
all of the aerosol material collected for at least 6 months by the
Batelle Massive Aerosol Sampler. These samplers shown in Figure
5 are capable of collecting over one gram of size fractioned aerosol
per day. The mass breakdown by stage of a typical collection is
shown in Table 11. The aerosol samples presently collected by the
Battelle Samplers are being characterized by ACB for epidemiological
studies. Since only two of these samplers are available at present,
this approach is impossible at this time.
5-A
-------
cr>
-. '. V.-—, SAMPLING SITES
Figure 1 LACS Site Location in Los Angeles
-------
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7-A
-------
Metj-
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Sampler inlets
3-4' above
-^freeway surfaceN
HT ..... - .....
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I )
y'
100'
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T
"A" site
CO Analyzer
Total Sulfur Analyzer;
•NO/NO- Analyzer
24-hr S02 Bubbler
24-hr Hi-Vol
4-hr Hi-Vol
4-hr Membrane
24-hr Cascade
Aiibient Temp, and Dewpolnt
Wind Speed
Wind Direction
Massive Aerosol Sample:
24-hr Dichotomous Sampler
0 Analyzer
B" site
""
24 hr,
4 hr,
24 hr,
Hi-Vol
Hi-Vol
Membrane
A
-<&
•f
Sepulveda Blvd
"110'
''C" site
CO Analyzer
Total Sulfur Analyzer
;iO/M02 Analyzer
24-hr S02 Bubbler-
24-hr Hi-Vol
4-hr Hi-Vol
4-hr Membrane
24-hr Cascade
'Massive aerosol sample
24-hr Dichotomous Sampler
83 Analyzer
"D" site
24 hr. Hi-Vol
4 hr. Hi-Vol
24 hr. Membrane
Wind Speed
Wind Direction
Figure 3 LACS Study Site Composition .and Elevation
-------
I HO"»l'(T AI. PNOTLCTJON A&lNCT
LOS ANOIES C*T*LT5T STUOT
Sl'LFATl
~~~I2S039IOI3
UG/CU MCTEH (25 C)
Table-!-
15/01/75
15/02/75
15/03/75
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15/05/75
15/06/75
15/07/75
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15/09/75
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15/1/75
IS/ 2/75
IS/ 3/75
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15 / 5/75
IS/ 6/75
IS/ 7/75
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IS/1 9/75
15/20/75 '
1S/2J/75
JS/22/75
iS/23/75
JS/2S/75
15/25/75
JS/26/75
JS/27/7S
iS/26/75
15/29/75
3S/3U/7S
15/31/75
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7-1
' " ' O.fl " ' '
5. 1
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l.s
4.S
s.7
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2.7
13. B
.22.5
4,79
26
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4.7
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7.6
|S«S*
1 3 « 9 o
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6*1
3*8
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• 2«6
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8
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-21.6
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1,34
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0.97
0,90
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Oi 46
1.17
1.07
0.7U
0.75
0.9|
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O.b1
s:
ss:
IS 3lUN|.rICANT AT P»0.9S
: IS SI&Nlr ICANT AT P-0,99
" ATYPICAL T'TMCY COUlO'BC QUESTIONABLE'
-------
LOS ANCLES CATALTST SluOT
LEAP (I5-I7J
""" ' |2|289201J"~~
UC/CU ritTEH (25 O
Table 2
... ..',:-• — SlIE-A ._ _S1IE B
05/01/75
OS/02/75
05/OJ/75
OS/OM/75
05/05/75
05/06/75
05/07/75
OS/Oo/75
05/09/75
05/10/75
05/1 I /75
05/12/75
05/1 J/75
OS/Js/75
05/ IS/75
05/16/75
C5/I 7/75
CS/18/75
05/19/75
05/20/75
05/2J/75
05/22/75
05/23/75
05/24/75
05/25/75
05/26/75
05/27/75
OS/28/75
05/29/75
05/JO/75
OS/Jl/75
AytHAtt
MAX,
H i N .
510, DtV.
TOTAL OBS
U.IO
0,30
o.io
O.JO
o.io
U.IO
0.60
0 • JO
' '•"* 0.10
O.bO
0.50
U.JO
"u.io "
0.20
0.20
0.2U
0.20
1.70*
"0.20
0.10
• "•" ' o.io
0. 70
O.JO
O.JU
0.10 "
o.io
- - - • • o.20
U.JO
0.20
O.J7
' ""' 1 .70 ""
O.IO
O.JO
. 2V
0.10
1 .00
0.70
0.60
0.50
U.JO
O.IO
I. HO
0.60
,0 . S 0 "
1 >00
o.oo
o.oo
o.oo '
O.IO
O.IO
O.JO
O.IO
1 .50*
0.50
O.JO
o.oo
0 .10
0.20
o.io
O.JO
0-20
0.50
O.JO
0.20'
0-50
1.50 "
0.00
O.B2
30
Sllt.C.
5.60
6.90
". 11,50
V . 10
•*—-•• l 6, JQ
7. JO
",'" 6.10"
. .6.30
V 5.90
JJ.JO*
n » 20
* v.iu
6.20
6.20
6 .00
6.80
J~ V.BO "
8-60
.8.50
6 «5U'
6. JO
6.00
5.6U
IJ.70
" io.«o
8>5U
" 7. JO
6*20
"" 6«VO
6'00
||.OU
7.«6
•--; u.jo -
5.60
2 « 1 V
31
• "'• " B(A "'
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... _siit. o.. ..—DELIA . _OE£I«_.
3.70
1.90
6.30
1.10
3.10
1.60
1.50
: 1.30
-— — -- ,( ..
_ .
6.20
~- " 1.00
3.60
. " 3.VO
1.70
. 5,00
1.30
1.90
I.JO
" 3.80.
1.00
" i.vo
65.80*
" 5.VO
I.JO
'"" 1.70
1.10
1.10 ""
S.JO
6.UO ""
O.UO
• v . ' ~" • '
o.id
c 0.50 ~"
0.20
-u.ov "~
0.00"
~: 0.80 '" ""
U.JO
0,10
0.20
"O.iV
•0.2V
"" -O.J9
-u.uv
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O.IO
-O.QV
2.bO*
O.JO
--O.OV
. -o.jv • —
-0.2V
"O.OV '
-U.|V
"-o.uv "'
. " O.IO
~ ' o. jo —
o.oo
"" ~u.oo
"6.7V'"" 0. I 1
45.80 " 2.80
3< 10 -0.19
~fl«5V 0.5V
28 29 •
5.20
V.|0
A. 20
7,00
5.70
»,20
5,30
|3.ou*
JU,80
6. 30
5.70
5.90
5 ,60
6.60
V.60
b,10
b, 30
1.80
6,10
5,60
5.20
I 1 .00
tO.so
«.20
6,90
6,|0
4.70
5.70
|U.80
7.10
|3.00
i.80
.IB
2V
OiA
C,H O.B O.C
..,.0[tI4......0CLlA _ DELIA _ __ UtLIi
3.30
1.10
3.00
1.30
1.10
1.20 .
... . t—
~ ~L ~t~-m ' •••
5.10
3.50
3.30
• — j.so
1.50
" "" 1.80 "'•'
1.10
~ 1,70 ~~
.2.60
"3.60
3.60
""'""""1.50
65.10*
. 5.60 '"
1.00
1.30
I.JO
" 1.20
S.OO
5.80 " "
• * 6.52
65.10
2.60
" JI.V7
26
5.20 3.30
I0.5Q
8.70
5.70
6.60
5.«0 ' ~
6. Ju
.50
1 .70.
1 .7u
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.iu - -
.20
,UU
6.70
V.7o
8. JO
8 . 10
2.UQ.
S.b'u
5.7u
5,6J
11. 30
10. 60 "•"•
U.IO
7.UO ~
6, DO
6.10
5.>U
10. bQ
7.J9
12.70
2,00
2.J8
JU
5.30
3.70
2. 50
1. |0
1.20
1.20
.
5«20
1.00
3.40
3.90
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• • 1 V
• JO
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65> 10*
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1.20
l.tl)
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3.70 ' •
5*00
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4.J9 .
45. SO
-O.lV
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27
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.1 . V9
• J. IV
.2.69
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-2.1V
• 2.bV
-2. 09
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bl.10*
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-2.5V
• 1.79
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51.10
• 5,1V
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2o
RATJO
CORR COEfF
St DirFERCNCE IS SIQNirJCANT AT P.O.95
ss: oirrcRENce is SIGNIHCANT *T P"0,vv
I. 35
1.08
0.64
2U.V2 '
l«.2lbS
•0,02
J8.07
2.765
0.21
IS. 52
I6.V5SS
•O.UJ
13.10
•0.01
U.b6
• U.1U
U.16
STARRCDI*) VALUES ARE ATYPICAL! THEY COULD BE QUESTlONABLI
-------
O
LOS ANGLES
PRO T t C T I ON «6lNCY
CATA.LTST
SUSPENOEO PART.
..... I | 10171 013
UG/Cu HEUR I2b C)
115-191
a -
T.eb]e»_l_.
O
Oi/OI /7S
OS/02/75
Oi/03/75
OS/CN/7S
OS/OS/ 75
CS/Ci/75
C5/C7/75
OS/OE/75
OS/09/75
OS/ O/.'S
CS/ 1/75
CS/ 2/75
OS/ 3/75
CO/ N./75
0 5 / ->/75
CS/ 6/?5
OS/ 7/75
OS/ 8/75
OS/ It 75
CS/2'j/75
CS/2 1 /75
0 '.• / 1 2 I 7 S
OS/23/75
OS/ZM//5
05/25/75
05/26/75
OS/27/75
OS/28//5
0 S / 2 V / 7 S
OS/ 30/75
05/31/75
.SilC.
97
116
" ' 91
67
112
68
92
••."' * S3
89
129
62
66
83
73
71
36
31
99
V3
88
75
62
80
93
78
106
83
1 13
1.. -SHE 6 ... SJ
" ' 107
135
130
107
52
1 18
77
1J1 •
17
90
• 113
73
83
93
79
79
1 1
• ~ 17
127
99
9b
89 ~
62
83
100
31
136
88
101
SI
— ' 6, A
•• O,A-
"c"»"
U.B
LIE.C - -SilElO— __OCLIA __OtClA OtLIa ULLlA ^OClIi.
151
173
182
192
1 37
112
137
123
169
127
1S9
133
137
113
137
135
96
111
<6B
1 18
|5U
13B
121
136
1 1fl
139
167
161
129
1 1»
|16
1 13 '
1 17
170
— 131
72
._. J22
106 .,
f
• 161 •
132
139
137
136
123
70
9b
1 16
150 '
117
Ib2
101
"ill
131
' ~" ' | 3 I
169
|13
121
- - ,j
10
11
13
-n
6
7
322
-5
1
11
II
17
10
6
.5
5
13
28
6
7
"11
0
3
7
3
30
5
-II
2
' 57
76
1J
15
25
55
• , 77
71
70
85
7|
71
• ' ' • 60
61
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60
80
67
..... ss
62
" '' ' 63
62
56
55
....... 4j
6|
— 76
16
67
H9
51
37
25
• — — jo. •'"
36
" *'
i
• * ~ ....
55
' 70
73
56
65
11 "• 17
51
61
17
" ' "* 57
57
.... . . j y
12 .
31
38
53
63
' — ' 60
• 8 •
11
i>
17
62
3U
6U
l'
16
-211
80
6V
71
60
5**
50
58
56
54
6/
1 1
l7
5*
1*
6*
53
is
5*
31
7i
2&
6>.
• ~ 37
11
10
21
10
' ~ " 1
27
• ~
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11
-- . s,
• 56
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59
11
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5 1
17
5 |
52
~ 63
12
i 1
31
' ' 50
33
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20
. HZ
o,c
uuu». .......
• 7
-27
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2
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1
* -II
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2
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2
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-7
• ii ' "
A v £ -< « 01.
81
100
132
18
61
17
MA* .
KIN.
STO.
TOTAL
*,'»
RATIO
T
COR«
S: D|
ss: o
otv.
06S.
— J
1
cocrr
Tf CRC
irrcR
129 ' •"'
31
23.08 65
27
• .__...._... .
N« IS SIGNIFICANT
ENCC IS SIGNIFICANT
11 - ~"211""
11 96
.32 21. VJ
30 31
-- •• •
AT P.O. 95
AT P-0.77
181 :""322""
90 -11
21.07 S7.18"
2b 27
U2H-
1.70
0.16
65 —
16
~|5.05 ""
27
I.7B "
22.ISSS
0,77
77
a
|7« 16 "
26
1.62 - ~
J1.77SS
0.71
• 60 "
• 2ii
'56. i 8
3-0
-: i.ii •
1.27SS
0.5J
' 63
1
|1.tt4
27
'I.3I
11.JVS3
0.81
•I I
-32
M.57
0.9 I
-b. J6SS
0.87
-------
NITRATE
1230692313
«25 C'l
Table 4.
05/01/75%
05/02/75
05/03/75
05/01/75
' 05/05/75
05/06/75
"Ob/07/75"
Ob/08/75
05/09/75
OS/IU/7S
05/1 1/75
05/12/7b
CS/13/75
OS/li/75
05/15/75
05/16/75.
Ob/ t 7/7b
05/1 8/75,
OS/1 9/75
Ob/20/75
05/21/75
05/22/75
05/23/75 • " .
OS/2H/75
OS/2i>/ >S. " ~"
Ob/26/75 '
05/27/75
Ob/2«/75. .
Ob/29/75
05/30/75
OS/Jl/75
AvERAQt
fl » Jt ,
HIN, .
sru. OEV» ""
T°T*^OBS. ....
RATIO 7* "
T **
CORR C&EFF
..SIZE. A
21.02
3.13
" 2.H8
B.iQ
21.67
10.25
1 1 .73
a, J7
"IB.UO
2 >. v 5
• 8.11
9,88
19.18
IU.6.1
1 o . d |
3. SI
2.10
6.17
""II .06
7.35
" 'l i . "<0
9.B7
1 i,b6
lb.27
' IU.52
16.38
B, V j
9.31
6.!>9
11.21
23.V5
2,10
~ "5,72
29
.....51U.B
23.12
21.70
3,7]
2. VI "•'
7.13
"22.06 "
10.07
ll.bb """"'
8,09
"-" 17.11
•__' 28 . 6 3 •
7 . 7 1
... _
10.1H'
."" " 11.25
3>65
1.87
,\ --
" "" )|.61 " '
7.6|
11-06
V 02SS
0,9«
O.A
.OEUI4...,.
0.90
O.bl
0.35
• 1 ,6B
.0,99
1 ""^
.3,19*
" 0.11
0,88
0,07 ~
.20
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,6b
•' ,16 •"
,bu
,24 ' '"
,15
" 0,65 "
• 3"* b 1 •
"' 0,62
i.i.i
.0.28 •
2,81
' 0,58
~ i *2»
1,19 —
0.15
.3.51
" ' | ,-11 —
26
t,:07 •••"
i .' !''%'& 0 '' ' '
0^9*
C,U O,B
.01118. ... ..DELIA .
"ru»l!5
3.9l»
•0.37
•U.62
0.^6
O.S9
1 «S9
1.36
' U.US
u . 2- 1
•O.I |
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I .08
2,25
0,3)
1*^9
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2.26
1*32
' 2.27 -
1 , *0
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2.2)
2.b7
1 «'9
- 2.27
•
1.24
3.VH
•0.62
I.U6
2?
' 1*13
4,275$
-1*18
2.27
0.30
0. | 1
l>32
•2* 07
•0.81
"
•8. | 7«
"1.11
.
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0.64
0.91
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• (9
•19
• .23
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• >27
• b6
• 10
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.72
0.31
2.27
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2.21
kb
0.79
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o,c -
• 0*>2
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U.6B
.' ~ 0 . 1 1 — ~"
0.36
.2.66 ~~~~
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0.02 —
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U.bB
O.bO ~'
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• U.|3
• 0.»3
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• U. 74
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• 1 . J6
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.U .il
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O.oS •
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i.B3 -;•- ~
'" — ..
u , v ) .— -
0.97
s: DIFFERENCE is SIGNIFICANT AT r
55! D1>FEKE*CE IS 3J&NIF1CANT AT.P*0.99 -" " .,'."
• I'" v"ALOCS"*'HE "ATYPICAL I'lHtT" COULO Bf QUESTIONABLC""7"" T-T—
-------
..->._...;.
CNVjMO.-'nCNTAL
LOS ANGLES
•
AMMONIUM
• • 123019
: UG/CU MLTE1 (25
'
1
05/01/75
_ 09/07/75
05/03/75
05/01/75
05/05/75
05/06/75
05/07/T5
C5/08/75
05/07/75
_ 05/1 0/75
" C5/ 1 1 //S
CS/ 1 2/75
C5/ 1 3/75
_ C5/M/75
C5/I 5/75
,_ 05/16/75
"05/17/75'
_ 05/18/75
C5/ 1 V/75
0 5 / 2 o / f 5
" 05/2J/75
C5/22/ ?5*
Ob'2j//S
05/2^/75
05/25/75
05/26/7S
05/27/75
05/2S/7S
" 03/29/75
05/ 30/75
~05/3|/75
AVt KAGL
" MAX .
• n IN.
570. DEY,
TOTAi. DBS.
XV w
RATIO i
T >
CORR COEFF
:• . . •
..^".^.sixc.t
0.10
0.21
0.21
0-21
0.03
• U.OO
O.UO
O.UO
- • u.oi
l.9|
U, 48
0.29
0,18
0. 30
"~ 0.10
0.06
"~ 0 . 1 2
0.29
O.UU
O.UO
U.OO
0.00
u. u
0>21
0.23
0.39
0.05
0.23
"u.U7't"
0.21
l.9|
u.uu
0.35
29
5HC.B...
o.m
0.9 J»
0>2|
0.23
0> IB
O.UU
O.UO
0.00
O.UO
0.00
• I .H6«
o.oo
_'.. 0'07
0.23
0.18
- O.UO
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•" u.uu
0.21
U,2| •"
0*05
0.19
0.21
. 0.00
0.00
0.12
0.30
0.21 "'
0.23
" 0. I 7
U. |9
1,16
O.UO
0.30
30
PROTECTION
CATALYST ST
1
2013
Cl
..sixt.c...
0.71
0.73
2>03*
0*53
0. t« .
0*10
. U'31
\0«29 .
0.16
0 • 3 1
O.UO
3> 1 2*
0.26
0>32
0>73
0,33
0*51
0.15
" ' 0 • 3 I
• 0> 10
0.39
0,3|
0,*6
0.30
0.3|
O.IH
u.^l
1,13
0.19
0«27
0«27
0>b1
3.12
0.00
0*60
31
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.
t,A
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U.31
U.32
0.27
0,27
.0.2H
0,29
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0.31
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. !•*!•_
.U.OI
0.03
0.25
0.03
0.1| .
0.09
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0.39
0.31
0,26 .
0.30
' 0,20
0,21
0.6tt
0,71
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0.20 -' '
0.28
1.21
-U.|8
U.26
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2,55
&.73SS
0,92
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..OCLXi...
0.31
0.25
0.22
0.13
Oi2B'
_ 0,16
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»U,27> *
.0.28*
0.02
0.02
0,|5
0,|9
0.33 ":
O.OI
0,17
0,29
0,13
0.25
0.18 ~~
0,20
0.31
0,12
0,21
0.03
o.oi '
0.16
0,58
.0.28
0.19
26
' 2.12
• , 7 i e«
0.90
•
C,H
' UELI&
U,l|
|.I2«
0.32
0.^5
0,3Q
0.3|
0.2V
1 U,U
0. J|
o.ou
| .66*
0*26
0./5
U.i>0
0. >b
u.5|
0.15
o.li
•0,13
o. IB
0,26
U.07
0.09
. O.i|
O.IB
0. 79
0.«3
O.^B
U.UI
" o.Jo
0.35
1 .66
• 0.13
0,35
30
2,»3
i'^SS
O.V2
0,b
nf i T i
0,31
1 ,1I«
0>25
0,20
Of |6
U.JI
0, 16
_ 1 tOJ*
• o.uu
• 0.06
U.27
0, |1
U,.2S
0.25
0. tb
' U.06
U. 26
U>21
0.21
U.UI
U.29
0.11
O.ib
' 0.2)
0.05
0.03
. ' -O.Ub
U.27
I'll
• O.UB
"J.3I
27
2.57
1.6ISS
U.92
•)
o,c
'. ' U.U3
O.ul
u.^v
-0.06
.O.oi
• U. 1 3
O.UU
-0. |2
-U.»2»
' -U.«5
• U. J|
• U«<2
O.UO
-U«*b
U.IO
O.ji
u./o
O.ou "" ~
• U.UI
0,17
• U.UI
• U.UI
-0.03
-0.33 ~'
-0.6 t •
• 0.22 ~ '"'
O.UO
" -U. 1 6
-0.08
0.29 •"
• 0.62
" il.il
2o
0.9| - •
-2.J3S
U.V3
5: ojrrcRCNCc is SIGNIFICANT AT P»o.9s
SSJ OJfFEHENCE IS SIGNIFICANT AT P'0.9?
"ST A^ £0 ( .r
A«E"" ATYPICAL! "THEY COULD' BE OUCST (ONAbLC ------
-------
1.
•V?
05/OJ/75
05/02/75
05/03/75
05/OH/75
05/05/75
05/06/75
05/07/75
05/08/75
05/09/75
os/io/;s
05/1 1/75
05/1 2/75
C5/I3/7S .
C5/ M/75
C5/I5/75
05 / I 6/7S
C5/I7/75
Ci'/ J 8/75"
05/19/75 -T
C5/20/75
05/2 1 /7&1
C5/22/75
05/23/7S
: 5 / ~ H / 7 1,
3-3/2 7," 7 5 ~
; 5 / 2 9 / •«
35/JO/75
,5/31/75. .
IV"*G£
tAX.
• TO. OtV.
•\.^.:'
*T>£JJ!"
o«« coirr
*"% ""' SVlf
^ (/ft/CV flCTI
31U 4 51!
e
|B
0
17
It)
30 •
8
0
8
21
0
t
6
2
3
3
18
... 25
It
21
_'_ IB
2
2
1 1
30
_ P
3> 28
• V
^ *
1210JVJ.OJ7 - •
CH I2S' CJ
[i a silt c '•• sue 0'.'
2H
26
6.
21
39
.. 15
16
30 '
26
17
13
• _; . • ._
8
1?
.... «7 ... _.„
31
3?
"•'" * ' ' 38 /' "" " "." "
30
..... . 28,'. ' ""' "" ,
' .. 2 J
;, ,15 .
23
1
4.
J • IU .
27
^ "
. . •
:._.. T.-._ •. . -.. .• ^
Table 6 ••;..'•'
DiA C,A DiA C,t» Ui6 0,C
"»6 ' ' '"•"
10
16 • ......
S
f ^ '
... / >« •_.''.•
.-- . " a -..-_. .._...._.
10
.... 17 .
»o '•.-...
"" It " ' '"• "' ' • •"• •' '
6 • i
16
13 •
M '••''.
^ .. .._........ ..,. .. •-..'- "
9
..•••' . '
: '•.- •"' 11 ' — .. — ' -. .--•.-.-.
;••;.'." . 23 • -. • . ••• • - • • '- ----- ^
1 • . •
. J3 ... . ... . . ....
*
•" " • :: 23 *""• ' -.- • — • •-
•••;•& ' , • .
_A_ <•> •_. "2*1 . . • . . • . _ . _. . .
'. i^'.loss
a,as .
is sicmricANi AT p«o.9s
IS SlONirjCANT »T >»0t»?
•*ftREO(») V*LUES"~/iRL~ATTPICAU"THET COULO 8C OUESTIONA8LI
-------
KONf INTAI. PROMOTION AGlNCY
LOJ ANGLES CATALYST 5TUOT
~ MONOXIDE
-.___. ^ CAR a ON
~" "' S2 101 I 1071
PARTS PER MILLION
MOURLT
Tabe 7
B,A
C,A
O.A
Oib
35/01/75
35/02/75
)5/03/75 '
)5/Ot/75
55/05/75
15/06/75
)5/0;/?5
35/07/75
15/10/75 ;_.-
35/ 1/75
jb/ 2/>5
:=/ 1/75
3S/ 5/75
05/ 7/75 .
05/ 8/75
0^/19/75
05/20/75
CS/21/75
Orj/:2/7S
05/TJ/75
05/2S/75
05/25/75
C5/2A/7S
05/27/75"
OS/23/75
05/27/75 '
05/30/75
05/J1/75
AVL^UE
HA X .
n 1 u .
STO. Otv,
TOT AC OB5.
1.5
2.6
0.7
J.7
3.7*
0.9
.2.2
1 .9
0.7
0.9
U-6
0.7
0*6
0*5
• 1
.8
.1
.5
.0
.0
0.5
0.7
0>6
0.7
0.3
1 >2
'3.7
0.3
0.78
30
3.1
H.S
H.6
2.5*
" 2.9
1.3
6.3*
'• M.9
1.8 -,_
6.1
6«H*
5>6
3.9 '
» 3.8
,S . 8
M.S
• "~ 3.8
•(••J ._
1.7
3.2
• 3.0
S.7
S.O
3.6 '
.-••H
6.S
2.5
0«95
29
j'.e . "
3.4 . V<.
' '•' . " "' '• ' 1.2 "" " .
•..:.,.«,
2'» . . . ' ' ' • .-;.
ill T. . ' •
.
5.0 ' .
3.2 . ,
3.2 .•',,.'
' ' "" '" " 1.H ' • '^
3.5 ;.)' . ' " '
: ' .'" " " '-2.o;-"' "V " " ~
2.8
.'. 3.6 " " ' ..'''.'•'' " . • •- -
3t7 .
' ' " 2.2 - ' ' ' ' - '
2.5
; .; " ;;;; . ' • •
3.6 •
T 3.3 . . " ^
3.0 • •• .
" ' 5.0 '
.0.5.
1.08 ' ' -
".28 . ....
tn
i
3; ClFFERtNCC'lS SIONIFJCANT AT P»0.95
ss: oifTEHCNce is SIONIFICANT AT p»o.99
~5TA^RtOV»T"V»LUtS "AHt." ATYPICAL" I" THEY cbuLb"BC
(I
-------
_ V-
L05 ASULCS CATALTSr STUPT
HOU«UT
. • '.
. .
05/31/75
05/02/75
05/03/-7S .
05/01/75
05/05/75
05/06/75
05/07/75
05/08/75
05/09/75
OS/.|'0/75
05/1 1/75
OS/J2/7b
OS/"l3/75
OS/1 N/75
05/15/75
U5/ 16/75
OV/'J ;/7.5 "
05/10/75 .
DS/Jf/75
35/20/75
15/71 /7S
S/i J/?5 "
J5/2N/7S
15/15/75
,5/26/75
15/27/75
!'s'/2'8/75
.'J/2V/7S
•5/30/75 '
5/3J/75 .
vLRjkGt
IN. '
TU« OEV.
3T*L 06S,
(^^
I T I 0
PARTS
,.S1.C-,..
0.12
O.J1
0.13
O.U7
0.05
0.02
0.02
0.00
0 .00
0.00
O.UO
O.OH
0.10
0.10
0.09
O.OH
0.00
u.oo
O.UO
0.01 .
0.03
u.oo
0,01
0413
u.oo
O.OH
22
1260 II 107 J . . . '"" " " " " ' —
PC* MILLION ; . . . - . . .
• *
..sin. D sm.c-....5iic.o.,
0,22
0.25
0.27 .
0*22
0,17 ' "
0.29
0.28
1 0»27
0.3)
0.27
0,17
0.29
o.32 •....;..
- 0.33 ""' *
0«3?
0.30 '
0.29 -1" """
0.36
0«27
0»38
0>32
' " "" '•" ". o • 2 a ' ' ' "~" ;
0.30
0.35 |
. , 0.30
0.17
6|A t»A 0»A C»k 0,8 P,C
.
. . ' '•
• • •
.." • • U.OB- ' . '^ • . _ v .. r. . _ .
0.20 ' •
Ll . 1 II
t ^ » 1 '~
fl.30 ' .
0.30 --...-.--
""-"•"• OJ33 -." * ~~ ~ " •
0,39
U, «»8
0.35
U.20 ' .
r ' 0.20
•--•--• ' 0.20 •'•' '" ' " "
0.. 3.t •
0.23
O..3b ""• ~ •" "
0.33
0,28 " •."-"- ' . .
0.27
u.35 ; '" •• • ~ j • • '• - ••- - •'•
0..2.B ' ......
0>0«
o.o; O.io • .
:;"- ' ' ': •' ••-••-•-;-- -,••_•-:-,,- -•-•-. 4. 7s •-/•.- -7-v ' • ' -• ••-
cocrr
>u.ss
.15 SIGNIFICANT AT pio.95
FERE Ncc'is SUN iriCANT AT'-.-P»O.?V
i •~>~TAlVC5~'AHE~ATTPiCAL'r THCV "COuLD"BC~QUCST IONABLT
-------
•^
Nftno&tN oioxloC HOURLY I . .
^
" S' 12602110M ....
• - PARTS PCR MILLION • . Table 9 • .....
>/C|/7S
i/02/75
1/03/75
1/01/75
. /O i/ 7 5
./07/75
./on/7s
, .' U ? / 7 5
./ 1/75
/ 7 / 7 c
,/ J/75
,/ 5/75
./ 6/75
i / 3/75
./ v/75
. / 2 0 / 7 5
,/2|/75
./22/75
./2J/75
1/21/75
•/26/75
i/27/75
./2B/75
i / 2 9 / 7 5 " "
>/30/75
. / 3 I / 7 S
(EKAGE
IX . "
rc. DCv.
)TAL OBSt
~^i .
JKR cocrr
: DIFFERENCE is
S ! OlFfEKE''CE I
'"'•-
0.01
0.03
0.01
0 >t)6*
0.01
0.01
0.02
0.02
0.01
0.00
0.02
0.02
O.U2
0.03
0.02
0.03
0.02
U.OI
0.01
U.03
U.03
O.U2
0.02
'0.06
0.00
O.UI
22
SIGN)
S SUN
.. B|A C,» 0»4 C,» U.d U,C
0.05 •
' • 0.05 .. .. -* . — . -
0.05 . • ' •
0«03« . ...'. .__. ... ; _..-,.- . - .......
0«03» '
0*05 , , ,. , -, - - — - .
0.05 ' .
0.05 ' r. — .. ..-. ,' .... — • •
""" " • 0-05 "»0l . .
o.'ai ' 0»oi ^ ... . _ .
0.05 . . • ' 0,01
0.05 0.00*
0.05
0«05 °«01 . . ... ._ . -
0.05 "'•' " ' J}«01 '
o»os- 9'03 -^ ...._' .. .
' " 0.06 """ "" W'01 • , • •
0.01 u«03
.. ^ o .05 U«OS
' -. 0.03 0.02 ... .. ... . _ '
• • •• - " • O.os 0.02
0.01 0.03 .
• — * - ' o.OI " ' • • 0.02
0.05 • _. _ .. y.-u2 ... . ....- ' . - —
1 '" ' ' o»U6 " ^'03
O.US 0.01 . .... .. . .
" 0«06 O.OS . .
0.05 . ' . V'01 _ . . . ._ . . ____
' 0.06 ' V 0.03
0»07« 0.05 ' •
— - - • 0.06 U.oi . ...
' O.U5 0.03 .
..._._ . o>07 .0.05
0.03 0.00 . . .
o.oo - 9«oi
31 22 . . _ .
-.'• — •' '• '1.96
|0.i|SS
u.o? ._. ... .. ... _ _.._ _.
MCANT AT p.o.ts .. '__ - - . - — .'....-.-- --• .-
IF1CAHT AT P»0t9? ~~ " •
— —
—
—
_ —
._ . '- j-
-
...
VALVES- ARE ATfPJCAL rTMET"cOyL'D BT'ouEST 10NABLE
-------
F-igure 4 .
HIND D-IRECTION(XFREQUENCY) RND flVG SPEED
BY SflflPLING INTERVRLtSITE R) >
05/75
07/11/75
6-10
8-11
3-7
C-S3
100.
35
60
o.
30.
SO.
sc.
52.
15.31!
3.0'
9.3 IE.
3.0 r
4.7 2 5.3 * . 4.7 X
3.3
44.
flVG KS
3.S PPH
9.0 r
4.7 S
5.0 HfH
2-0 X 0.7 X
72.3X
14'.; v.
•>. o-o x 0.7 •/. O.T x 0.
5-6 fl?H
. 1
' 0-7
40.
CO.
-*. I;. 4 >.
". „ I PUT
. 1 >.
d_^i_^
4-t-'
N , lit
9.7
.-) X
SF . . S , " SH
•-NI, ' '
-------
Table 10
HOURLY VALUE LOGOUT.ROUTINE
YEAR
1975
1975
1975
1975
1975
19:7^1-
1 97-5^ c
1975
19<75-e
1975
1975
1975
1975
1975
i95H:tc~
1975
1975
1975
1975
1975
1975
1975
DAY
314
314
3 1 4
3.1.4
3 :l 4
314
314
314
314
315
315
315
315
315
- "315
-nctf'15
315
315
315
315
315
31'5
315
315
TIME
1 5 : 00
16: 00
1 7 : oo
IB: oo
1.9:00
20:00
21 :oo
. 22:00.
23:00,,
00:00
01 :oo
,* .02:00 f
. V>3:ob"
04:00
^ti5-{^)~0v-
l67YooJ
< 108*00 (
09:00
10:00
1 1 tOO
12:00
13:00
14:00
NORTH BOUND
05815
07308
07461
07064
06110
04244
03094
03218
02846
02314
01338
00797
004J.O
00309 •'•-
-r^-.c 00352" "
''""' U'"03:tV6
-;..--•: 7 :0509 7 ri^.ss)
05163. '
O53T49.-.1
05204
04992
05153
SOUTH BOUND
04625
05060
. 04850
04925
04184
03294
02081
02265
02194
01370
00797
00399
00281 v.
00281
- * •-' v -00433* ,. :«.
, ...... .01781 ^ . . -
~ H '''\ '06891 •-•-•«'--•-•
~ •'« i ;,.-:, 1078' IS -'• --^ '.:.
07631
06635 5.655 f.
05675
04868
04149 ""
04795
TEN MINUTE VALUES PER LANE
.*.)
HOURLY VALUE LOGOUT ROUTINE
r
r
YEAR
1975
1975
1975
'1973'
1975
1975
1375
1975
1975
1975
1775
1975
1975
1775
1975
1975
1975
1975
1975
1975
1975
1975
1975
1975
DAY
315
315
315
'315
315
315
315
315
315
315
315
315
315
315
316
316
316
316
316
316
31.6
" 316
316
31'6
.TIME
10:00
ll'.OO
12:00
1 1:00
15:00
16: 00
17:00
IB: oo
19:00
20.: oo
21:00
22:00
23: oo
00:00
01 :oo
02:00
03: oo
04:00
6 s: oo
.06: oo
-------
; \ -
Figure 5
Scolping stage,
cut off * 20/i
JlJUXlU
••:7I;
3.5/1 stager
stage
Electrostatic
precipitotor
20-A
-------
Table 11
PARTICULATE COLLECTION AT L. A. SITE
Site Location Si
Site A
3.
i.
< 1.
zc Fraction
5-20
7-3.
7 urn
p.m (1st
5 |itn ( 2 nd
stage)
stage)
(electrostatic
plates)
Tine I'criod
~ A
~ A
- A
days (100
days (100
days (100
hr)
hr)
hr)
Total
Site C
(downwind)
3.
I.
< 1.
5-20
7-3.
7 urn
p,m (1st
5 \im (2nd
stage)
stage)
(electrostatic
plates)
~ A
~ A
~ A
days (100
days (100
days (100
hr)
hr)
hr)
Mass
1
0
3
6
l
0
A
.AAO
.8A5
.730
.015
.2A5
.9A5
.A95
g
g
g
g
g
g
Total 6.685 g
21-A
-------
DETERMINATION OF HUMAN BODY BURDEN BASELINE
DATA OF PLATINUM THROUGH AUTOPSY TISSUE ANALYSIS
by F. Vandiver P. Duffield,* Anna Yoakum,** Joseph Bumgarner,* and
John B. Moran*
Results of analysis for platinum in 97 autopsy sets are presented.
Analysis was performed by specially developed emission spectrochemical
method. Almost half of individuals studied were found to have detected
platinum in one or more tissue samples. Deposition of platinum in 13
of 21 tissue types is reported. Surprisingly high values are observed
in subcutaneous fat, previously not considered to be a target site for
platinum deposition. These data will serve as an estimated human tissue
burden baseline for platinum in EPA's Catalyst Research Program.
*Environmental Research Center, Environmental Protection Agency, Research
Triangle Park, N.C.
**Stewart Laboratories, Inc., 5815 Middlebrook Pike, Knoxville, Tennessee
-------
Measurement of Platinum from Catalyst-Equipped Vehicles, Combustion
and Attrition Products
Mr. John Sigsby
Environmental Sciences Research Laboratory
Environmental Protection Agency
Research Triangle Park, North Carolina 27711
Notes used in presentation (attached).
24-A
-------
PRODUCTION CATALYSTS 1975-1976 CARS
ro
en
i
MONOLITHS
BASE MATERIAL
CORDIERITE ( 4 C M G F E 1 0 ' 4 AL2 0 3'10 Si 02 ' H20)
R I L A R I T E (K20 '4CAO'2AL203'24Si02'H20)
WASH COAT Y - A L U M I N A
CHLOROPLATINIC ACID (H 2 P T C L 6) ADDED
PACKED BED
BEAD AND EXTRUDED PELLETS
Y ALUM I NA-.H I GH PURITY
-------
CATALYSTS
PACKED BED - GM - AMC
PO
CTi
TWO SI.ZES 160 C LI, IN, - 1 8 " L 0 N G - V E G A AND ANC (6CYL.)
2 6 0 C U , IN,* 24 "LONG-ALL G M BUT VEGA
%
B.E.A D, S, . AND PELLETS INT E0R C H A NrG E A B L E
* 3 KG OF PACK I NG
A M C. D I F F E R E f! T F 0 R H U L A T I OJ! .THAN 6 M
.,0.5 TROY OZ , ( 1 , 56 GRAMS )
-------
CATALYSTS
ro
ONOLITHS
FORD-ENGELHARDT ( 6 0 % ) - P T / P D AND PT
f-1 A T H E Y BISHOP ( '4 0 I ) - P T / R H - 1 9 7 5
PT/PD - 1976
-0,05 TROY 0 Z , ( 1 , 5 6 G M s ) ME T A L
CYLINDER 5 " D I A , - 1 3 " LONG
ALSO USED BY MANY FOREIGN M A N U F A C T U R E R S ( V W E T c, )
CHRYSLER - UOP
V A R I A B L E - C 0 MP 0 S I T I 0 N AND LOADINGS
OVAL 8 " WIDE -15-18" LONG (MUCH LARGER
THAN ABOVE FORD)
-------
ANALYSIS OF TUBE SWEEPINGS
CO
I
Sample
A
B-E
F
F
F
Laboratory
EPA-ESRL
EPA-EMSL
SWRI
Stewart
Livermore
Method
Wet AA
Wet AA
Flame! ess AA
Activation Analysis
Emission etc.
AA
ESCA
Pt vg/gm
«
245 - 635
60 - 145
0.034
2.44
0.165
100
West Pt+4
-------
F i Iter
Air
in
Engine
C y n G ~prr,e t e r
f\j
vo
cng i
Figure 2
Flow Dioqrom for Engine Exhoust
Porticulote Collection
Air
out
I ns trimen t
and
Control Room
•„ . . ^
?/ i x i n g
i fe
Part iculote
rav i r,2 tr i c Fallout
Flow —>-
Control
i ! i >
4
i
O
^i V i. /
\
$
\ /
\ 1 /
Scmpl ing Slits
- — ' oil Pipe
N
\,
X
~s
\
\
^>>
"X
Anderson _^
Seporctor
"•»•
•'•—S tender d Muffler
/^
i
19 . S°0tt R=seorch ins, Killinor^
•v
^
•^
™ ond ,'02 F.lter
Anclys i s
Ai
| Pu
i
y
l_
F isher Gas Port it i oner
CO, C02, N2, 02
Beckmcn 109A
Total Hydrocarbon
Analyzer
Exhaust P ipe
Flow Meter —>
Vacuum
• Pump
n
•o
O
I
Kenoneter
-------
SNAIL. P ARTICULATE EMISSIONS
SOURCE
EXXON
EPA-RTP
DETECTION LIMIT
< 1 0 " 7 G M / M I L E - < 1 0 ' 5 M G R / c M -
X-RAY AND CHEMICAL
•
< 1 0 ~ 5 G M / M I L E - < 1 C " LI u G R / c M - X-RAY
oo
o
1 , 5 6 • G R A M S TOTAL LOADING - 50,000 MILES OPERATION
1,56 G M
50,000 MILES
= 3,1 x 1 0"5 G M/M I L E
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lexicological Studies of Auto Exhaust Catalyst Components
Dr. Jerry Stara
Environmental Protection Agency
Cincinnati, Ohio
The attached reports were used as the basis for Dr. Stara's
presentation.
31-
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B.4
Comparative Effects of Intratracheally Administered
Sulfuric Acid ar.d Palladium Sulfate on Pulmonary Free Cells
K. I. Campbell
E. L. George
T. Grumbles, and
Y. Y. Yang
SUMMARY
The comparative acute toxicity of sulfuric acid (H^SCM) and palladium
sulfate (PdS04) to pulmonary free cells was investigated in rats using the
intratracheal route of administration. These materials were of interest
because of their potential emission as constituents of exhaust from
automobiles using high-sulfur fuel and noble-metal catalytic devices. The
pulmonary free cells (PFC), normally primarily alveolar rr.acrophages, are an
important component of pulmonary defense and clearance functions.
H2S04 solution was administered in a dose of 0.45 mg [S04~2j/kg body
weight to one test group (5 rats each group) and an aqueous solution of
hydrous PdSG/i was given to the other test group at a [SO/j-Zjdose rate
equivalent to that of the H2S04 group and [Pd^jdose of 0.5 mg/kg. The control
group received deionized water (H^O) only. Dose volumes ranged from 0.1 to
0.14 ml per ret, and body v/eights averaged 188 gm. One day later the PFC
werg collected by saline endobronchial lavage. Total and differential cell
counts, cell viability, phagocytic activity, and lung and body weights were
determined.
With respect to several criteria, there were marked and significant treat-
ment effects in the FdS04-exposed animals but not in those exposed to H2S04.
Noted effects included the following: increased total numbers of free cells;
increased sub-populations of both viable and nonviable nucleated cells,
phagocytic and nonphagocytic macrophages, polymorphonuclear leukocytes, and
red blood cells; increased numbers of phagocytized test-spheres; increased
dried lung mass. There were no apparent effects on proportions of cells
nonviable, on proportional phagocytic activity (spheres per cell), or on
cell size (diameter).
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The results indicate definite effects by PdSO/; on the pulmonary free
cell system suggestive of a recruitment of alveolar macrophagr:S and/or
retarded cellular clearance activity, of leukocytic infiltration, and of
hemorrhage and perhaps cdc-r^a. These effects are regarded as .'having toxic
implication for palladium compounds (especially soluble and ionizable)
gaining access to the lungs' terminal air spaces, and 'being associated
with the cation moiety rather than the anion [SO/j'^Jor acidity of the material.
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B.5
P75-31.6
EFFECTS OF VARIOUS METAL SULFATES on SUCCINATE-DEPENDENT RESPIRATION
S. D. LEE, M. RICHARDS, L. MCMILLAN
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI/ OHIO
R, SHELLEY, V. N, FINELLI
UNIVERSITY OF CINCINNATI
CINCINNATI, OHIO
For Presentation at the 68th Annual Meeting of the
Air Pollution Control Association
Boston, Massachusetts June 15-20,1975
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75-31.6
UTECrs OF VARIOUS MJ7TAI, SULFATES ON SlA-acn Monitor wis used to measure .the
O2~uptake. V.'c tested the effects of various sulfatcs, such as cadriur1.,
palladiin;., rar.grmeeo, r.?.crnesiur.,- calciii:', sodium, and iuncniurr. on the system.
The results indicated that the sulfate icn in tissue slices or in hcnocenatc
did not affect the respiratory chain. However, ancr.g the cations, Cd4^
appeared to be the most potent inhibitor. Cacri'-m inlubited the 02-uptaXe by
approyjurately 501 at 2 x 10~7 H and 100; at 3.3 x 10~5 M. Othsr catioj^.s did
not shov: iriiibitorj' effects at sini laj- concentrations. To achieve a JC*
inhibition by Pdr/D^ noire than 10"^ K w?.s reruircd. Cadmirn ion, a kno-.n
potent inhibitor of the nitcchonJrial rcs:;iratcry chain, was utilized in this
opprijncnt as a reference to>:icr.nt. CdSO^j V.TIS found to ho manyfold rore
toxic to the respiratory chain than KSO4. Other cations, such as KnJ~1', ivq"t~f,
Ca4^, were not inhibitory at oor.centraticns up '_o 10-3 M, but appeared to have
a slight stinulatcry effect. Ce4"*"1", r:a+, ?r.d 13'.^ did not cause any effect.
Intragastric acninistration cf a single and nultiple dose cf CdSC^, PttS^Jj.
,£ind PdSO4 at 3 ?jnd 60 tr-ole per kg body v.vi^ht, did not affect succir.ate-
depcr.dont respiration in any of the organ tissues tested (kicr.ey, liver, heart,
and lung). It is probable that, under these experimental conditions, these
rrotals were not absorbed in significant arotmts to cause any effect en the
mitochonirial respiration of soft tissue orcans. The in vitro o^rirL-nent
suggests tJxat at least Cd^"1", the nost potent inhibitor tested, should have
impaired this cnzyne systen if absorbed in significant quantity and accumulated
in the tested organs.
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INTERACTION OF METAL BINDING AGENTS FROM
COMBUSTION PRODUCTS V.'ITH TRACE METALS
L. V7. Michael, V. N. Finelli, V. J. Elia, H. G. Petering
Department of Environmental Health
Kettering Laboratory, University of Cincinnati
Cincinnati, Ohio 45267
and
S. D. Lee and K. I. Campbell
National Environmental Research Center
Environmental Protection Agency
Cincinnati, Ohio 45268
ABSTRACT
Metal binding agents present in the environment may react with trace
metals thus altering essential biological functions. Combustion pro-
ducts from tobacco, gasoline and coal as well as from other pyrolytic
processes are ubiquitous sources of such material. We have demon-
strated the potential for such interactions in biological systems by
comparing the copper binding activity of condensates from selected
sources.
The reactivity with other metals was also examined. The isolated
metal binding fractions have been tested for effects on metalloenzyraes
and mitochondrial respiratory activity. In addition, the efficacy of
*
this concept was tested in vivo using a chelating agent, 3-ethoxy-2~
oxobutyraldehyde • bis(thiosemicarbazone), (H-KTS). H-KTS has been
found to alter cholesterol metabolism In a manner similar to that ob-
served in copper deficient rats.
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CHEMICAL CHARACTERIZATION AND BIOCHEMICAL TOXICITY
OF ORGANIC AND INORGANIC COMPONENTS OF AUTO EXHAUST
Contract No. 68-03-2011, Project No. Cl-7^-C217
Annual Progress Report - June 20, 197*4 to June 20, 1975
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A. INTRODUCTION
This is the first annual report on research progress
achieved betv;een June 20, 197^ to June 20, 1975 carried out
under contract No. 68-03-2011, Project No. CI-7^-0217 of NERC/EPA,
Cincinnati. The research data reported herein'^are the result of a
coordinated and collaborative effort betv;een this laboratory and
ETRL project Officers, Si Due Lee, Ph.D., and Kirby I. Campbell,
D.V.M.
During this year we have achieved substantial progress in
the following studies:
1. We, have characterized the auto exhaust gas emissions
(AEGC) from engines with and without catalytic converter as to
titratable acidity, copper binding activity and total organic
carbon.
2. We have isolated from the whole exhaust condensates the
protonated and non-protonated ligand fractions and tested these
metal-binding fractions as inhibitors of tyrosinase, a copper
containing metalloenzyme.
3« We have assessed certain .biological effects in rats
exposed to irradiated and non-irradiated auto exhaust emissions
and to single components of auto exhaust, such as carbon monoxide
and sulfuric acid mist. Lipid metabolism, serum and liver lipids,
hematologic effects and organ damage were the parameters studied
in the exposed animals.
4. We have investigated the effects of noble metals, plati-
num and palladium, and of other heavy metals, cadmium and mercury,
on 6-aminolevulinate dehydratase (ALA-D) of rat erythrocytes, a
zinc and -SH containing enzyme, and on succinate-dependent respir-
ation of rat lung, kidney, heart and liver homogenates. The effects
of the above toxicants have been evaluated and compared in these
metal sensitive systems.
5. We have initiated nutritional studies to evaluate the
effects of components of exhaust emissions on essential trace
metal metabolism and to determine the protective roles that cer-
tain micro-nutrients may play against the toxicity of these agents.
on A
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H. BIOCHEMICAL STUDIES
Exposure studies were conducted at the ETRL/NERC facilities in
Cincinnati in collaboration with Dr» S.D. Lee.
1. Biological Effects of Auto Emissions
V7e have participated in a series of studies designed by ETRL,
"Toxicity Assessment of Mobile Emissions" (TAME). The purpose of ,
these experiments was to determine the efficiency of oxidizing
catalysts in reducing certain pollutants. The qualitative and
quantitative alterations of emissions due to the introduction of
an oxidizing catalytic converter to the automobile exhaust system,
along with the possibility of introducing nev; pollutants such as
platinum and palladium in the environment, called for toxicity
studies in animals exposed to engine exhaust emissions. A series
of exposure experiments, whose characteristics and conditions are
reported in Table I, were conducted to evaluate the toxicity of-
irradiated (I) and non-irradiated (N-I) exhaust emissions from
engines with and without converter. Other variables in these
experiments were the length of exposure, the sulfur content of.
the fuel and the type of engine. The animals used in the first
experiments(TAME1s I,J, and K) were Sprague-Dawley lactating fe-
male rats. In the remaining experiments, adult male Sprague-
Dawley rats were used.
To assess the effects of a catalytic converter on the toxicity
of engine exhaust emissions, two pairs of exposure experiments can
be utilized: TAME I vs.. TAME J and TAME 0 vs. TAME N. The first
pair of experiments was conducted to determine the effects in
lactating female rats of short term exposure (7 days, 2^ hours/day)
to emissions from an engine with and without converter, TAME J
and TAME I, respectively. The other, pair of experiments, TAME N
and TAME 0 were designed to compare the effects in adult male rats
exposed to the emissions of an engine (1975 Ford, UOO CID)
equipped with a converter and tuned as specified by the manufacturer.
The conditions in both experiments were kept the same with the exception
that in TAME 0 the converter was removed from the exhaust train.
In these two latter experiments the animals were fed a semipurified
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diet with controlled levels of essential trace minerals,
especially copper and zinc (Table II), to study the impact of engine
emissions on certain aspects of the metabolism of these trace
metals. The other experimental animals v;ere fed commercial chow.
The data collected from the various experiments are reported
in Tables III through XVII. An overall analysis of the results
suggests that various biological parameters have been significantly
altered in the animals exposed to non-catalytic emissions (TAME I
and TAME 0 experiments). However, some slight alterations and
a trend of higher serum cholesterol were also noticed in some
of the rats exposed to catalytic emissions. In summary, we have
observed the following changes:
a. As shown in Table III, TAME I animals (lactating female
rats), both I and N-I groups, showed a dramatic loss of body
weight. This effect was more severe in the I group. In TAME J.
the exposed animals also showed a loss in body weight, although
the effect was less pronounced in this study than in the preced-
ing one. In TAME K the animals exposed to the engine emissions
gained more weight than the control animals. No significant
effects on growth were observed in the other TAME studies, except
for the rats exposed to the non-catalytic emissions in TAME 0
where a significantly slower growth was evident in N-I and I groups.
The growth patterns of the animals in the first 3 TAME studies
cannot be compared with those of the remaining experiments since
two different animal models were used; lactating rats were utilized
in the earlier studies while male rats were chosen thereafter.
Rats used in TAME N and TAME 0 seemed to show better growth
patterns than the others. This observation may be explained by the
fact that the animals in these studies were fed a semipurified diet
which met all the NRG requirements in both macro and micro dietary
constituents (Table II).
b. Elevation of serum lactate dehydrogenase (LDH), which
may indicate possible tissue damage, was observed in I and N-I
groups of TAME I. No effects were present in the experiments
where a catalytic converter was used (Table IV).
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c. In all the experiments serum glutamate-oxaloacetate
transaminase (GOT) was normal in the exposed animals. This obser-
vation, along with the elevation in serum LDH in TAME I animals,
may suggest that the target organs of toxic components of.non-
catalytic emissions may be the lung and/or trie, kidney, probably
excluding the heart and the liver.
d. Serum lysozyme was measured only in TAME J and TAME "K.
An elevation of serum lysozyme activity would have indicated a
possible effect on the lungs. However, the exposed animals in
these experiments did not show any alteration of this parameter.
Analysis of urine lysozyme in the same groups of rats were not
conclusive, since the values obtained for controls and exposed
animals we.re scattered and yielded a large' standard deviation.
A significant increase of lysozyme .activity in the urine of -the
exposed animals would have suggested kidney damage .•
e. Blood analyses showed that hematocrit and hemoglobin-
fables VIII and IX) were significantly-elevated in the animals
exposed to non-catalytic emissions (.TAME I and TAME 0). A "poly-
cytemic response (Erythrocytosis^) >may have occurred in these
animals since the levels of CO in the exposure chambers were
elevated. The increase in hematocrit was not due to'dehydration
since serum albumin was found to'be^normal-in the exposed animals.
f. The effects of engine emissions on lipid metabolism, as
shown in Tables X through XV, can be summarized as follows: N-I
and I groups of TAME .L sho.wed 'an- elevation of serum triglycerides
which may be statistically significant.
Animals in TAME N and TAME 0 had higher serum triglycerides
than those in TAME M. This effect may be due to the dietary change
(semipurified diet vs. laboratory chow).
There seems to be a trend of higher serum cholesterol in animals
exposed to either catalytic or to non-catalytic emissions. How'ever,
this effect is not. statistically significant except in TAME N where
N-I and I groups are statistically different from control, p<0.01.
There is no difference in total liver lipids and liver phospho-
lipids between exposed and control animals. -' However, 'liver 'chol-
esterol seemed to be affected in TAME 0 where the value for the
I group is significantly lower than that for control.
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The in vivo incorporation of acetate-1- C into liver lipids
van not affected in any of the exposure studies.
g. Serum zinc and copper v;ere analyzed in the experimental-
animals of TAME N and TAME 0. In the animals exposed to non-
cntalytic emissions serum zinc was elevated ^'-concurrently serum
f
copper decreased in the sarne animals. No alterations of these
parameters were noted in the animals exposed to catalytic emissions.
h. Erythrocyte 6-aminolevulinate dehydratase (ALA-D) was
substantially elevated in the I and N-I groups of TAME 0. ALA-D
is a zinc dependent enzyme involved in the synthesis of heme. This
elevation may be due to an alteration in the metabolism of zinc
or to the 'presence in the exposed rats of more immature red blood
cells which may ccntain a higher concentration of ALA-D.
2. Effects of Carbon Monoxide Exposure
Four groups of 20 adult male Sprague-Dawley rats were
exposed to atmospheres'containing various concentrations of carbon
monoxide (CO) for periods of 2 .and 4 weeks. The carbon monoxide
levels in the four exposure chambers were 0, 57-5» 172.5* 517-5
mG/m (0,50,150, and U50 ppm). The following parameters were
measured: body weight, lung, liver, kidney and heart weight,
serum enzymes (GOT, LDH, and HBD), hematocrit, hemoglobin,
serum albumin, serum cholesterol and triglycerides.
a. Gross biological effects — In the animals exposed to
the various levels of CO for 2 and 4 weeks there was no significant
effect in body weight gain when compared to the control rats.
Serum GOT, LDH and HBD were not affected in any of the groups, thus
confirming that CO exposure, at these levels and conditions, did
not cause cellular damages in rat organs such as liver, lung,
kidney and heart (see Appendices 1 and 2). From the organ weight
measurements, we found that the heart was the only organ affected
by CO exposure. There was a significant increase in the heart
weight of animals exposed to CO (Figure 1). Our results agree
with those reported by Theodor et al. (J. Occup. Med. _13:2U2, 1971)
These authors attributed the cardiac hypertrophy to an increase
in cardiac work due to an elevation in blood volume and viscosity
(High hematocrit).
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b. Effects on hematocrit. hemoglobin and serum albumin-- An
elevation in hematocrit and hemoglobin levels was observed after
?. weeks of CO exposure (Figures 2 and 3). A follow-up study on-
hcnatocrit was performed after termination of CO exposure. Figure
*rshows a drastic increase in hematocrit during CO exposure and a
•f
p.radual return to normality during the following recovery period
of 'I weeks. Serum albumin was not altered in the exposed rats.
( c. Effects on serum cholesterol and triqlycerides — There
nppcars to be a trend to lower serum cholesterol values in the
CO exposed animals which may be significant only in the group
exposed to the highest level of carbon monoxide (Table XVIII).
Ho significant effects were evident on serum triglycerides in
the animals exposed to any concentration of CO (Table XIX).
Furthermore, during TAME 0 experiments we exposed a group of
six rats fed semipurified diet to CO alone. In this study the '.CO
levels were kept similar to those observed in the whole emission
i o
exposure chambers, that is, 230 - 46 mg/m (see TAME 0 aerometry
data). The results reported in Table XX show that hematocrit and
hemoglobin in the CO exposed animals are substantially higher than
control. Other parameters affected in the exposed animals were
ALA-D activity, serum zinc and liver cholesterol. Again, as in
the previous experiment, we noticed that the CO exposed rats
showed a slight reduction in serum and liver cholesterol.
Elevations in hematocrit and hemoglobin (erythrocytosis) were
reconfirmed in this study. An increase of ALA-D activity in the CO
exposed animals may be explained either by the alteration of zinc
metabolism, since this enzyme is Zn-dependent (Finelli et al.,
Biochem. Biophys. Res. Comm. 60:1^18, 197^5 see Appendix 3),
or by a possible increase in number of immature red blood cells
which may contain a higher concentration of ALA-D.
3. Effects of Sulfuric Acid Exposure
In TAME M and TAME N studies, run concurrently with exhaust
emissions exposures, two groups of adult male Sprague-Dawley rats
o
were exposed to H0SO.. mist, 6.59 mg/m° for 90 days and of 6.35
o d. "i
mg/m for 28 days, respectively. Growth rates, hematocrit and
hemoglobin levels were not altered in the animals exposed to
HgSOjj (Table XXX). Tables XXII, XXIII, and XXIV show the effects
of fUSO^ exposure on the lipid metabolism. Of the various para-
meters measured only the level of phospholipids seemed to be
significantly altered. The liver phospholipids in the H
-------
exposed croup of TAME N v/ere 6 6$ higher than control. However,
no significant alterations v/ere noticed in the in vivo incorpor-
ation of acetate into liver lipids. •
A. Effects of Intraqastric Administration of Pt, Pd and Cd salts
on SuccJnate-Stiniulated 0 -uptake
Adult Sprague-Dawley' rats (300 to 400 g) were exposed to
ulnglc and multiple intragastric injections of platinum, palla-
dium and cadmium chlorides and sulfates. In the single dose exper-''•
Incnt v;e used two concentration of the chlorides (40 and 80 jamols/kg
body weight), and the animals were killed 24, 48 and 72 hours after
exposure. Succinate-stimulated Op-uptake was tested in the homo-
gcnntes from liver, heart, lung, and kidney (Table XXIV). No pro-
nounced effects were observed in the rats exposed to 40 and 80
/imols PtKpClg after 24 and 48 hours post administration. Cadmium
chloride seems to enhance Op-uptake in liver, heart and kidney-
nt both concentrations, 72 hours post-injection. This effect
could be explained by a possible uncoupling of the mitochondrial
oxidative phosphorylation. A depression of Op-uptake in lung homogena
wns observed with CdClp 24 hours post-exposure; however, this effect
decreases at 48 and 72 hours, incubation time. Further studies
using isolated mitochondria may be required to confirm these
findings arid elucidate the mechanism by which Cd affects the
mitochondria electron transport. In vitro studies conducted by
other investigators and by us have shown Cd to be one of the most
toxic metals affecting the succinate dependent respiration.
In the multiple-dose experiments groups of rats received
daily 8 and 80 juunols of the compounds per Kg body weight for
4 consecutive days. The animals were sacrificed 50 hours after
•
the last exposure. Succinate-dependent respiration in the homo-
genates of the various organs was not significantly altered by
any of the compounds used (Table XXVI). These findings suggest
that intragastric administration of Pt and Pd salts do not affect
the mitochondrial function of the cell in various organs. The
reason for the lack of detectable effects may lie in the low
intestinal absorbtion of these compounds (see W. Moore, D.
Hysell, W. Crocker and J. Stara in Toxological Studies of
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I'al Indium and Platinum Document,
5. Effect of PdClp Injected Intravenously into Rats on
* t_
Succinate-stimulated 0 -uptake in Tissues
*~~ "~ t_
We measured the succinate stimulated respiration in homo-
eenales of'liver and heart tissues of rats which had been in-
jected bi-weekly with various doses of PdCl?. The total dose, of •
PdCl in saline solution received by five experimental rats
consisted of 6.5 mg/kg body weight injected in 7 portions (3
Injections of 0.5 mg, 3 of 1.0 mg and 1 of 2 mg/kg of body weight).
The control group (5 rats) received at the same time injecticns
of saline solution. Four weeks after the last injection the
animals were..sacrificed', and the liver and heart were homogenized.
The O-up'.take-values for the experimental and control groups
are, reported in .Table'XXVIl1. l There "was no significant -effect
ofiRdClp exposure on either heart or Driver 0 -uptake.
6. 'In-.Vitro Effects of Various^ Metal's on (S-Aminolevulinate
Dehydra ta'se">-'.Act-i'Vi-ty '-
:The. following^compounds were tested in vitro on the erythro-
' .. i ....-..•..'. ' • ' '
cyto - 6-amlnolevuMrr.a-te de'hytrata!se '(ALA-D) activity: HgCl2,
ClUHgGl', PtgCLjij, "HgPtcTg-1, CuCl'2,"znCl2", and PbClg. Lead ion is
a known inhibitor of this enzyme; thereforev we compared the
effects of all the"1 other metals to the inhibitory capacity of Pb
At concentration'" 6-f ' 10" M PbClp there was-approximately 50$
....*- • '.-
inhibition of ALA-D activity. Le'ss inhibitory were CuClp, HgCl ,
•- -. •> • ' > •'•.«: • 2
and CdC'l0 which produced the same extent" of inhibition at concen-
^ -c; -h . "i" •• '
trations:>ra-1ging from 10 '^ to 10 M. These metals v;ere 10, to 100
times .less inhibitory than lead. The least ALA-D inhibition was
r - ..•••'
produced by :.the( following'compounds : 'PtK Cl^, H PtCl^-, PdCl_,
and CHQHgCl. This group of compounds 'produced 50^ ALA-D inhibition
J . -l\ _^j
at concentrations ranging from 10 to 10 M; that is, they.are
100 to 1,000. times less active than lead. " Zinc ion has been found
to activate ALA-D.' The in vitro effects Of various metals on
ALA-D are reported in Figures 5 and' 6.
We had previously shown that ALA-D is a zinc dependent enzyme
(Table X%VII) and now have observed both in vivo and in vitro
I i »^ . .,•.'• •
interaction of Zn _, and .rPb .; on the activity of this enzyme
(see Appendix ^).
•\-»'
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We have plotted erythrocytc ALA-D activity versus time
7) for two groups of rats fed a semipurified diet low
In zinc aMd supplemented with Zn in deionized drinking water • '
(?.5 and ^40 ppm, respectively); from the seventh to the ninth
week one half of the animals from each group deceived 200 ppm
j 1 *
Pb in drinking water.
In all animals there was a dramatic loss of erythrocyte 'ALA-D
activity with aging. The animals in group II, which received
a lov/ zinc diet, showed considerable lower enzymatic activity
than those in group I, (statistically significant, p^0.05)»
The addition of lead to the drinking water caused a reduction
In enzymatic activity to zero in the low zinc group and almost
to zero in the zinc-fortified group. Following the removal of
the lead exposure, ALA-D activity in rats fed the higher zinc
level returned to normal values more quickly than in those en '.
• X
lower zinc diet. Five weeks after withdrawal of lead the enzyme
Activity in the lead-treated low- zinc group was still significantly
lower than in the control group, whereas there appeared to be no
significant difference between lead-treated and control groups in
the, animals fed a higher zinc diet.
The effects on erythrocyte ALA-D activity by in vitro addi-
tion of ZnClp to blood obtained from rats fed low and high zinc
diet with and without lead exposure are reported in Figure 8.
In all cases zinc reactivates the lead inhibited enzyme; at
10~ M ZnClp ALA-D values are similar in lead-exposed and control
animals.
In another experiment, rats were fed for one month laboratory
chow containing 500 ppm lead (as lead acetate). Two months after
withdrawal of dietary lead the average level of lead in the blood
of these animals was about 50 jug/lOO ml and ALA-D activity ranged
between 10$'and 20^ of normal. Figure 9 shows the activation of
ALA-D in the blood of one of these animals by the in vitro addition
-U
of ZnClpj again we noticed that ALA-D activity peaked at 10 M ZnClp,
Flcure 10 show the ALA-D activation by ZnCl0 in adult rat blood
-S
with and without the in vitro addition of 10 -'M PbCl0. Between
_h _c - 2
10 and 10 -'M. ZnClp, the enzyme activity reached about 150$
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i.
[ of control in both incubation mixtures.
i The data lend further support to the reports that ALA-D
IP A zinc-dependent enzyme. The return of ALA-D activity to
: f.orr.nl after termination of lead exposure o'ccured more slowly
i
In tlio low zinc group. The inhibition of ALA-D by lead may
therefore be due to an antagonistic effect of lead on zinc
rotnbolism resulting in a functional impairment of this enzyme. .-,
These findings may be of great importance as a means of
Improving plumbism therapy and in determining the exposure level
of the toxicant in relation to the nutritional state of the
exposed population.
C. Chemical Studies
1. Collection
a.) Sources
Chemical studies of auto exhaust were conducted on
materials condensed from a 1972 Chevrolet V-8 350 CID engine
operated at 30 MPH steady state (collection sequences I through
IV), and a 1975 Ford V-8 400 CID production "California" engine
(all subsequent collections). The latter engine was equipped with
an Engelhard monolith catalytic converter on American Lava substrate.
Runs V and VII through X were conducted at 20 mph and run VI at
25 niph all at steady state. Engine performance data for the
collection series are shown in Table XXIX. Collections were con-
ducted at ETRL/NERC facilities in Cincinnati under the direction
of Dr. Campbell.
b) Traps
A system of two consecutive traps (A and B) were
employed for trapping auto exhaust gas condensate (AEGC). The
first trap (A) was cooled with water and ice and the second trap
(B) was cooled with a slurry of acetone and dry ice. A third
acetone scrubber (C) was used in some collections. During run I
and part of run VI traps A and B contained acetone.
Samples were collected adjacent to the converter on the pre-
treatment side (engine side of the catalytic converter) for non-
catalytic exhaust (NC) and the post treatment side for catalytic
exhaust (C).
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2. Chemical Parameters
a) Total Organic Carbon
Total organic carbon was determined with a Dohrmann
Rnvirotech Organic Analyzer, model DC-50, on AEGC as shown in
t
Tnble XXX, as a measure of material collected.'. In general, the
cnrbon content of AEGC was greater in NC-AEGC than C-AEGC as
anticipated. Large variations were observed between collections.
Results in this report are reported relative to the volume of
exhaust passed through the collection system as reported by
ETRL/EPA Cincinnati in Table XXIX.
b) Apidity
The acidity of AEGC was measured by titration with sodium
hydroxide and by pH measurements of the condensate as received.
These data are shown in Table XXXI. The titration showed two
readily identifiable inflections. These were indicated by differ-
ential plots of the titration curve, pH/ vol. NaOH versus
volume of NaOH. Catalytic AEGC was approximately three times as
acidic as NC-AEGC. The slope of the titration curve is indicative
of 'a buffering capacity in addition to a high concentration of
acidic capacity functional groups vrhich is consistent with the
low pH of the condensate.
c) Copper Binding Activity
1. Extractable Copper
The ability of AEGC to effect the distribution of copper
between aqueous and organic phases (Michael, et al. Environ.
Sci. Tech. 5:2^9, 1971) was used as an assay for determining the
efficiency of exhaust collection. The range of values reported
in Table XXXII suggests that comparisons between collection periods
or other collection variables must be made with caution.
Wet and dry trapping systems shown in Table XXXII, refer to
the presence or absence of an acetone scrubbing solutions in the
sample collection system. The presence of a scrubber increased the
trapping efficiency as determined by extraction.
The results of extraction studies show in general that more
copper was extracted by NC-AEGC than by C-AEGC. These results are
based on liters of exhaust passed through the collection system.
49-A
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The extraction procedure is a simple or one stage equil-
ibration between the immiscible solvents in the presence of
excess copper. Thus, this technique does not reflect the amount
of material v:ith potential total dynamic interaction in a system
w»»ore the subject material is in continuous coatact v;ith fresh
reitcrial, which is. the usual case in vivo. What has been
demonstrated is the formation of complexes which are capable "of crossing
A phase barrier. Even if these are ion pairs, the transport
across the phase boundary shovis the potential for forming lipid
soluble metal binding complexes or neutral complexes which might
be able to cross a cell membrane.
«
ii-. Total Copper Binding Activity
Total copper binding activity was determined,by .measuring
the copper stripped from a carboxymethylcellulo,se .ehromatographic
columns in the Cu (II) form,... (Finelli et,al., Environ. Sci. Tech.,
6:7^0, 1972). The.total activity represents the copper eluted by
both ethanol and ammoniac a,! eth.an.pl. In general the-catalytic
exhaust bound, more copper.,. The variations between, runs was
greater than the difference between the total Cu. binding-..capacity
of O and, NC-AEGC as shown in Table XXXIII.
iii. Fractionation of AEGC
Condensates vrere transferred to A-meithyl-2.r-p.entanone (MIBK)
solutions by treating with 2,2-dimethoxypropane and expess>fIBK
and reducing the.rvolume ,of,aquepus solution. The MIBK.solutions
of AEGC were fractionated ,on the. CMC-Cu columns, which were . -.
described, in the above section. • Ethanol had previously .been .shown
to elute copper binding compounds with:displaceable protons. -•
Non-protonated ligands which fornucharged complexes with Cu (II)
were displaced by anunoriiacal-'e.thanol eluent. Fractionation data
are., summarized in. Table , XXXIV,.
Complexing agents with, displaceable protons predominated in
both NC- and C-AEGC.. Catalytic AEGC's greater total copper rbinding
activity is attributable to complex-ing1-agents with displ-aceable.
protons: The level-of neutral complexing^agents-was about the
same in .paired collections of C- and NC-AEGC.
d. Enzyme Inhibition Studies
The effect of fractionated and non-fractionated C- and NC-
AEGC, of run vn,on the activity of tyrosinase (Finelli, et al.,
50-A
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7ox. Applied Pharm., 27:*415, 197*0, a copper enzyme, has been
Investigated. The fractions tested include the non-metal
binding fraction, F, , the complexes eluted from CMC-Cu columns
with ethanol, F , and those eluted by ammonial ethanol, F,..
• - » . O
Ihccc fractions v;ere stripped of copper on CMd-,HT column .to
yield metal free fractions, F^, F^, and Fg respectively. The
Kinetic studies of tyrosinase yielded typical noncompetitive
inhibition graphs when reciprocal velocity was plotted versus
reciprocal substrate concentration. The activity of fractions
and nonfractionated AEGC' is shown in Table XXXV.
The differences in activity between the activities of copper
complexes of run VII C-AEGC follows that anticipated for fraction
F_ followed by the metal free fraction F(-J namely, the inhibition
of the free complex, FJ-, is greater than the metal bound complex^
5 •
F?. The unanticipated inhibitory activity of the F^ fractions -of
both C- and NC-AEGC is attributed in part to the same trend observed
in blanks; namely, an inhibitor is being eluted from the CMC-Cu
column.
The differences in activity of nonfractionated AEGC and the
fractionated material suggests additional interactions which are
unknown. A material balance is required to elucidate this problem.
c. AEGC Stability
Attempts to correlate collection, fractionation or enzyme
activity have been frustrated by the lack of an adequate monitor
or assay for a material balance. Copper binding activity varies
with time. Although samples could be stored under refrigeration,
when samples were maintained at room temperature the copper
binding activity increased. Both total and extractable copper
binding activity increased. Therefore, the interpretations and
interrelationships between collection sequences must be made
«
with caution.
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! D. Slr.nl Ticr-nce of Results
1. Biological Studies
The in vivo studies on the biological effects of engine
enlr.cions have shown that an oxidizing catalytic converter was
very effective in preventing in exposed rats loss of body weight,
or^nn damage and hematologic changes all of which were observed
In rats exposed to non-catalytic emissions. Some of these
adverse effects, such as elevation in hematocrit and hemoglobin
levels and cardiac hypertrophy, were due solely to the high
concentrations of carbon monoxide in the exposure chambers,
while other, effects, such as impaired growth and organ damage
no shown by elevation of LDH in the serum, must be attributed
to other components of automobile engine exhaust' emissions. In
conclusion these results indicate that the introduction of a
xatalytic converter in the engine exhaust train not only elimin-
ated the deleterious effects of carbon-monoxide, but also the
effects of other toxic constituents of engine emissions not
presently identified.
The in vitro studies have shown that platinum and palladium
i
compounds, important components of catalytic converters and
possibly new pollutants, seem to be less toxic than other heavy
metals such as cadmium and lead when tested on succinate-dependent
respiration and ALA-D activity. However, these studies alone
t , ; * ' *
are insufficient for concluding that platinum and palladium are
relatively non-toxic and therefore not a threat to the
environment. .
Dietary zinc has been found to'be protective against the
in vivo inhibition of ALA-D by lead. This Pb-Zn interaction has
been shown also in vitro. These findings may"be of great import-
ance in assessing the exposure-level of the toxicant in'felation
to the nutritional state of the exposed population. Furthermore,
zinc, administered with the chelating agents now used in plumbism
therapy, may be more beneficial in lead detoxification treatment.
Serum zinc and copper have been found altered in the animals
exposed to nori-c'atalytic emissionsi therefore an in vivo inter-
action between these essential metals and engine exhaust
52-A
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pollutants has been established. Carefully planned nutritional
•tudics may be an aid in the studies of subtle effects of
pollutants.
2. Chemical Studies
An interaction betv;een AEGC and copper has been shown by
tho formation of copper complexes. A portion of these complexes
could be distributed between iraniscible solvents indicating
thnt neutral complexes were formed. The in vitro demonstration
of synthesis and mobility suggests the potential for biological
Interaction.
Fractipnation of C- and NC-AEGC showed that compounds with
dir.placeable protons comprised the major portion of the ligands
which formed copper complexes. This is consistent with the
presence of an appreciable level of titratable protons in
AEGC samples.
Studies of tyrosinase activity show greater inhibition by
C-AEGC than NC-AEGC. The copper binding fractions of C-AEGC
nlco showed greater inhibition of tyrosinase activity than
NC-AEGC.
. 53-A
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Biological Fate of a Single Administration
of 1!"Pt in Rats Following Different '
Routes of Exposure
W. MOORE, JR.. D. HYSELL.
W. CROCKER. AND J. STARA
U. S. l-Mvirniiiiienltil Protection Agency. Office nf Research IJeveliipiiieni.
National l-'.nvironnicnial Research Center. Environmental To.ricnlti.vy Research Laboratory-.
Cincinnati. Ohio
Received May 6. 1974
The rclcnliim. tissue clislribulion. and excrelion of l!"Pt in ;idull rats was determined fol-
lowing oral, intravenous (IV). and intratracheal administration. The highest retention was
obtained following IV dosing, and lowest retention (less than O.S^f-) occurred after oral
dosing. Tissues containing the highest concentrations of ""Pi following IV administration
wore I he kidney, adrenal, spleen, and liver. Following a single oral dose: almost all of the
""I1! was excreted in the fcccs due to nonahsorplion. whereas after IV dosing, similar quan-
tities were excreted in both the urine and feces. Following IV dosing of pregnant rats. '"Pi
was found in all the fetuses: however, (he amount was small.
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Preliminary Studies on the Toxicity and
Metabolism of Palladium and Platinum
*.
by W. Moore, D. Hysell, L Hall, K. Campbell, and J. Stara
Preliminary data a IT given on the Ml-- of PdCI- following different routes of exposure
and on (lie 1,1) of PlCI, following inlra*cuoiis exposure.
The retention, (issue distribution, and excretion of '"'Td and "'PI in rats was deter-
mined following oral, in! ravenous, int ratraclu'al, and inhalation exposure. The highest
retention for holh '"'I'd nnd "''Pt was ohtainrd following intravenous dosing, and the
lowest retention occurred after oral dosing. Following n single oral dose, almost all of the
'''I'd and '"'Pt was excreted in the feces due to nonabsorplion, whereas after intravenous
dosing, similar <|iianlilies were excreted in holh the urine and feces. Tissues containing
the highest concentrations of these metals were Hie kidney, spleen and liver. Following
inlrateiioiis dosing of pregnant rats, a small amount of '"Td and ""Pi was found in the
fetuses.
* U.S. Environmental Protection Ajrency, National
Environmental Research Center, Environmental
Toxicology Research Laboratory, Cincinnati, Ohio
46268.
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B.I
WHOLE BODY RETENTION IN RATS OF DIFFERENT 191Pt COMPOUNDS
FOLLOWING INHALATION EXPOSURE
W. Moore, Jr.,* M. Malanchuk,* W. Crocker,*
D. Hysell,* A. Cohen,* and J. F. Stara*
(This manuscript has been submitted for publication in
Environmental Health Prospectives)
* U. S. Environmental Protection Agency, National Environmental
Research Center, Environmental Toxicology Research Laboratory,
Cincinnati, Ohio 45268
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The whole body retention, excretion, lung clearance, distribution
<
and concentration of 191Pt in other tissues was determined in rats
following a single inhalation exposure to different chemical forms of
191Pt. The chemical forms of 191Pt used in study were 191PtCl4, 191Pt(S04)2i
191PtO and 191Pt metal. Immediately after exposure most of the 191Pt
was found in the gastrointestinal and respiratory tract. Movement of
the 191Pt through the gastrointestinal tract was rapid with most of the
l9^Pt being eliminated within 24 hours after exposure. Lung clearance
was much slower.with a clearance .half-time of about 8 days. In addition
to the lungs, kidney and bone contained the highest concentrations of
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B.2
INHIBITION OF HYDROXYPROL1NE SYNTHESIS BY PALLADIUM IONS
Rao S. Rapalca,* Keith R. Sorcnsen, Si Duk Lee** and
Rajcnclra S. Bhatnagar***
Laboratory of Connective Tissue Biochemistry
Scliool of Dentistry, 630 Sciences
University of California
San Francisco, California 94143
^Present address: Albert Einstein Medical Center
Northern Division
York and Tabor Roads
Philadelphia, Pa. 19141
**Dr. Lee's Address is: Health Effects Research Laboratory
Environmental Research Center
•—•"" ' Environmental Protection Agency
Cincinnati, Ohio 45268
***Author to whom reprint requests should be sent.
61-
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SENARY
Palladium ions, administered as PclSO^ markedly affect, tlic
incorporation of L- (3,^-^IL) -proline into non-dialyzablc fractions
in 10-day chick embryo cartilage explants with a 55-65% reduction
in the concentration range 0.06-0.6 jnM. Under these conditions
the synthesis of 3H-hydroxyproline was nearly completely inhibited.
Experiments with prolyl hydroxylase indicated a strong irreversible
inhibition of the enzyme with a.competition between Fe2+ and, Pd"+.
The Ki for the inhibit ion-was 0.02 mM.. , Tliese studies suggest tliat
Pd?+ may inhibit, collagen synthesis by replacing Fe2+ in the
active site of prolyl hydraxylase. .Tliese studies., also point to a
potential mechanism of Pd2t toxicity.
i 62-A
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13.3
PALI ADI IJM LNTF.RFERL7S WITH CONNECTIVE
TISSUE mCRQMOLECULAR SYNTHESIS
63-A
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ABSTRACT
Palladium is a component of catalytic converters installed in auto-
mobiles to reduce hydrocarbon coicentrations. There is a liklihood that
Pd may enter the environment as a result of attrition and disposal of
such devices, however, little information is available on its toxicity.
Studies reported here suggest tliat Pd^* ions may be highly toxic to con-
nective tissues and cause inhibition of macromolecular synthesis, if
these ions reach the target systems.
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B.4
Comparative Effects of Intratracheally Administered
Sulfuric Acid and Palladium Sulfate on Pulmonary Free Cells
K. I, Campbell
E. L. George
T. Grumbles, and
Y. Y. Yang
SUMMARY
The comparative acute toxicity of sulfuric acid (H2S04) and palladium
sulfate (PdS04) to pulmonary free cells was investigated in rats using the
intratracheal route of administration. These materials v;ere of interest
because of their potential emission as constituents of exhaust from
automobiles using high-sulfur fuel and noble-metal catalytic devices. The
pulmonary free cells (PFC), normally primarily alveolar macrophages, are an
important component of pulmonary defense and clearance functions.
H2S04 solution was administered in a dose of 0.45 mg [S04~2J/kg body
weight to one test group (5 rats each group) and an aqueous solution of
hydrous PdSC4 was given to the other test group at a [$04-2]dose rate
equivalent to that of the H2S04 group and [Pd+2] dose of 0.5 mg/kg. The control
group received deionized water (^O) only. Dose volumes ranged from 0.1 to
0.14 ml per ret, and body weights, averaged 188 gm. One day later the PFC
were1 collected by saline endobronchial lavage. Total and differential cell
counts, cell viability, phagocytic activity, and lung and body weights were
determined.
With respect to several criteria, there were marked and significant treat-
ment effects in the PdS04-exposed animals but not in those exposed to H2S04.
Noted effects included the following: increased total numbers of free cells;
increased sub-populations of both viable and nonviable nucleated cells,
phagocytic and nonphagocytic macrophages, polymorphonuclear leukocytes, and
red blood cells; increased numbers of phagocytized test-spheres; increased
dried lung mass. There were no apparent effects on proportions of cells
nonviable, on proportional phagocytic activity (spheres per cell), or on
cell size (diameter).
65-A
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The results indicate definite effects by PdS04 on the pulmonary free
cell system suggestive of a recruitment of alveolar macrophages and/or
retarded cellular clearance activity, of leukocytic infiltration, and of
hemorrhage and perhaps ederra. These effects are regarded as .'having toxic
implication for palladium compounds (especially soluble and iohizable)
gaining access to the lungs' terminal air spaces, end being associated
with the cation moiety rather than the anion [SO^Jor acidity of the material
66-A-
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B.5
EFFECTS OF VARIOUS METAL SULFATES ON SUCCINATE-BEPENDENT RESPIRATION
S. D. LEE, M. RICHARDS, L. MCMILLAN
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO
R. SHELLEY, V, N. FINELLI
UNIVERSITY OF CINCINNATI
CINCINNATI, OHIO
For Presentation at the 68th Annual Meeting of the
Air Pollution Control Association
Boston, Massachusetts June 15-20,1975
67-A
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75-31.6
ITTECTS OF VARIOUS MCTAL SULFATES ON SIKXBJATE-DEPE.'DRTT RESPIRATION
S. D. Lee, H. Richards, I,. 1-fcMillan; FPA, Cincinnati, Ohio
K. Karaffa and V. Finelli; University of Cincinnati, Cincinnati, Ohio
The use of catalytic converters to control hydrocarbons and carbon
monoxide in the autcmobile exhaust emissions resulted in an increased output
of sulfatc(s). This, along with possible oiussions of noble metals from the
converter prcrpted us to test conparative toxicity of the various sulfates in
an enzyne system. Succinate-depcndent respiration w.is tested in rat liver
slices incubated in Krebs-Rinqer phosphate buffer and in liver horogenate
incubated in 0.25M pliopphatc buffer, pi! 1.4, A Clark-type oxygen electrode
attached to a YSI Model 53 Biological Oxygon Monitor was used to measure .the
O2-uptake. Ke tested the effects of various sulfatcs, such as cadmium,
palladium, rar.gnnese, ragnesium, calcium, sodium, and arnonium on the system.
The results indicated, that the sulfate ion in tissue slices or in hcrogenate
did not affect the respiratory chain. However, anor.g the cations, Cd++
appeared to bt the most potent inhibitor. Cadr-ium inliibited the O2~uptake by
approximately 50% at 2 x 10~7 M and 100% at 3.3 x 10~5 M. Othsr cations did
not show inhibitor^' effects at sinilar ccnccntrationE. To achieve a 505
inhibition by PdSO^ more than 10"^ M was rerruircd. Cadnium ion, a kna-.n
potent inhibitor ct the nitcchondrial respiratory chain, was utilized in this
operiment as a reference toxic.-'.nt. CdSOfl was found to he manyfold more
toxic to the respiratory chain than PdSO*). Other cations, such as Mn4"1", t-'q**,
Ca"1"*;, were not inhibitory at concentrations up to 10-3 M, but appeared to have
a slight stinulatory effect. Ce+^f, Na+, and 13'.^ did not cause any effect.
Intragastric administration of a single and mltiple dose of CdSO^, Pt(SOjj)2,
.and PdSO^ at 0 and 60 unole per kg body weight, did not affect succinate-
dependent respiration in any of the organ tissues tested (kidney, liver, heart,
and lung). It is probable that, under these experimental conditions, these
metals were not absorbed in significant amounts to cause any effect on the
mitochondria! respiration of soft tissue organs. The in vitro experiment
suggests that at least Cd4"1", the most potent inhibitor tested, should have
impaired this enzyme system if absorbed in significant quantity and accumulated
in the tested organs.
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INTERACTION OF METAL BINDING AGENTS FROM
COMBUSTION PRODUCTS WITH TRACE METALS
L. W. Michael, V. N. Finelli, V. J. Elia, H..G. Petering
Department of Environmental Health
Kettering Laboratory, University of Cincinnati
Cincinnati, Ohio 45267 " .'
and
S. D. Lee and K. I. Campbell
National Environmental Research Center
Environmental Protection Agency
Cincinnati, Ohio 45268
ABSTRACT
Metal binding agents present in the environment may react with trace
metals thus altering essential biological functions. Combustion pro-
i
ducts from tobacco, gasoline and coal as well as from other pyrolytic
processes are ubiquitous sources of such material. We have demon-
strated the potential for such interactions in biological systems by
comparing the copper binding activity of condensates from selected
sources.
The reactivity with other metals was also examined. The isolated
metal binding fractions have been tested for effects on metalloenzymes
and mitochondrial respiratory activity. In addition, the efficacy of
•
this concept was tested in vivo using a chelating agent, 3~ethoxy-2-
oxobutyraldehyde • bis(thiosemicarbazone), (H-KTS). H-KTS has been
found to alter cholesterol metabolism In a manner similar to that ob-
•
served in copper deficient rats.
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B.7
PUU-DNAKY BIOCHEMICAL 7\LTERATIONS RESULTING FROM OZONE EXPOSURE
Mohaivniad G. Mustafa
Center for the Health Sciences
University of California School of Medicajie
Los Angeles, California 90024
Si Duk Lee
National Environmental Research Center
United States Environmental Protection Agency
Cincinnati, Ohio 45268
ABSTRACT. 1-iatabolic response of lung tissue to ozone was studied in rats
and monkeys after exposure of animals to various levels of ozone (0.1 to 0.8
ppm) for 1 to 30 days. In rats, 0.8 ppm ozone exposure resulted in a 40-50%
augmentation of oxygen utilization in lung homogenate in the presence of an
added substrate (e.g. succinate or 2-oxoglutarate). Activities of marker
enzymes, viz., mitochondria! succinate-cytoclirome c reductase, microsomal
NADPH-cytochrome £ reductase and cytosolic glucose-6-phosphate dehydrogenase,
increased maximally (40-70% over control) after 3 to 4 days of exposure, and
remained elevated throughout the continuous exposure for 30 days. In monkeys,
the observations were the same except that the magnitude of biochemical
changes was relatively smaller. Exposure of animals to lower levels of ozone
resulted in proportionately smaller biochemical changes in the lung, and
ozone effects were detectable for up to 0.2 ppm level. While 0.1 ppm ozone
exposure was ineffective, dietary deficiency of vitamin E, a natural anti-
oxidant, increased the sensitivity of rat lungs to this concentration of ozone.
The results suggest that low-level ozone exposures may cause metabolic
alterations in the lung, and that dietary supplementation of vitamin E may
offer protection against oxidant stress.
71-A
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The Use of Chemical lonization Mass Spectrometry for the Analysis
of Platinum and Palladium.
Terence H. Risby, Department of Chemistry, Pennsylvania State
University, University Park, Pennsylvania, 16802.
The preparation of the l,l,l-trifluoro-2,4-pentanedionates of
platinum and palladium will be presented. This procedure can be
used for the direct analysis of Pt or Pd in unknown samples.
The chemical ionization mass spectra for these complexes will
also be reported together with the predicted mass spectra based on
the natural isotropic abundancies of the metals, 0 , C13 and H2.
The C.I. mass spectrometer used in this study has been modified
so that solutions of the chelates can be injected directly into the
mass spectrometer.
Preliminary sensitivity studies suggest that the limit of
detection for these complexes is of the order of 1 x 10~12 g.
73-A
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Chemical Transport of Platinum
from Automotive Catalysts
A Laboratory Study*
John Hawley
New York State
Department of Environmental Conservation
Laboratory tests with a noble metal beaded oxidative automotive catalyst
indicate chemical transport of platinum. Samples from the exhaust gases
of a furnace employed to house and heat the catalytic reactor were
filtered through glass and quartz fiber mats which were then subjected
to x-ray fluorescence analysis. Scaled to a 60 mph cruising automobile,
particulate platinum in microgram per mile quantities was collected
during some experiments. Ethylene dibromide (5 ppm) and carbon monoxide
(3%) as additives and air and nitrogen as carrier gases were studied in
all combinations in the final experiments. No platinum transport was
induced, even at elevated temperatures, with pure nitrogen carrier gas,
whatever the additive. Otherwise, oxygen was the most important
component in determining platinum transport from aged catalyst samples;
bromine enhanced transport in the presence of CO, but depressed trans-
port when alone at higher temperatures.
Parallel engine test stand experiments with exhaust converters containing
the same catalyst material showed microgram per mile platinum transport
with ethylene dibromide added to the fuel and reactor temperatures
raised via modified timing and carburetion. Earlier engine experiments
at lower temperatures showed no transport either with or without ethylene
dibromide.
This report was submitted in fulfillment of EPA Grant No. R-801486 by
the Division of Air Resources of the New York State Department of
Environmental Conservation under the partial sponsorship of the
Environmental Protection Agency. Work was completed as of July 1, 1975.
Based upon the Final Report on EPA Grant No. R-801486 by R. D. Harris, R. E. Gibbs
and N. Kolak.
75-/1
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NOV 2 4 i9/b
Platinum Metals in Air Particulates Near A Catalytic Converter Test
Site As Measured by Isotope Dilution SSMS*
J. A. Carter and W. R. Musick
Composite dust samples weighing about one kilogram were supplied by
Dr. R. Thompson, EPA-RTP, for Pt, Pd, and Ru analyses. The origin of the mate-
rial was airborne dust particulates collected by filter banks which were located
about 1.4 km from a catalytic converter test facility. The sample was a blackish-
gray mish-mash of particulate matter. The major constituents of the ashed com-
posite (50% wt loss) were Si, Al, Pb, Ca, Fe, Na, Zn, K, Mg, and Ti. Many minor
constituents, including Pt, Pd, and Ru may be determined by excitation with radio-
frequency spark followed by spectrometric measurement of the mass resolved ion
beams. This technique of spark-source mass spectrometry when applied to the
composite was not sufficiently sensitive; only Au was detected at the 1 ppm level.
Therefore, to obtain adequate sensitive and quantitative Pt, Pd, and Ru results,
we dissolved a large sample so we could equilibrate enriched stable isotopes of
Pt, Pd and Ru, with each of these isotopically unaltered elements present
in the samples prior to platinum metal enrichment as a metal precipitate. Gold
was added as a carrier. Gold and the platinum group elements were concentrated
into the Au precipitate which was subsequently used in the isotope dilution spark-
source mass spectrometry measurements.
Recovery of the Au by weight was near theoretical for blank acid samples,
but was always less than 50 percent for the sample solutions. This fact did not
alter the validity of the results, however, since equilibrium was established prior
to the precipitation process.
The results by isotope dilution spark-source mass spectrometry in picograms
per cubic meter of air are as follow: Pt-2.5, Pd-0.5, and Ru-<0.1. These results
assume an average air burden of 50 yg M .
*Work supported by the Environmental Protection Agency under EPA Interagency Agree-
ment D5-0466 with ERDA. ORNL is operated by the l.friion Carbide Corporation for ERDA.
By occeptDnco ol thio nrtlclo, tho publl .FIT or
recipient Pcknowl.-oVos iho U.S. Government's
rlcht to retain a non - exclusive, royalty - (rao
license In and to any copyrleht covering the
article.
-------
Measurement of Pt and Pd in Ambient Air, Soil and Water
Dr. Donald E. Johnson
Methods for collection and storage of these samples will be
presented along with the analytical methods. The assay methods
emphasize atomic absorption with a graphite furnace.
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Measurement of Pt and Pd in biological Tissues
Dr. Donald E. Johnson
Methods for collection, storage, sample preparation and
analysis will be presented. The limits of sensitivity for each type of
sample will be given.
80-A
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Platinum Analyses in Animal Tissues and Fluids
* * . **
Andre F. LeRoy , Walter S. Friauf , Charles L. Litterst ,
** ** *
Theodore E. Gram , Anthony M. Guarino , and Robert L. Dedrick
Introduction
Interaction of platinum and its complexes with biological systems
has become a matter of increasing interest and importance. Individuals
may be exposed for therapeutic purposes in the treatment of cancer and
through environmental contamination from vehicular emission control
devices or some industrial processes.
We have adapted flameless atomic absorption spectrophotometry
(AAS) to the quantitative determination of Pt in biological tissues
and fluids down to ~30 ppb. The methodology has been .applied to
observation of the distribution and disposition of the anti-cancer
drug cis-^PtC^CNH^^J > DDP, in several animal species following intra-
venous administration.
Analytical Considerations
Therapeutic doses of DDP are of the order of 1 mg/kg; so, analytical
methods must be capable of quantitating platinum at concentrations less
than 1 ppm. Samples from animals subjected only to environmental
exposure would be expected to contain much less, e.g., <2 ppb .
*Biomedical Engineering and Instrumentation Branch, Division of
Research Services, National Institutes of Health, Bethesda, Maryland,
**
U.S. Laboratory of Toxicology, National Cancer Institute, Bethesda,
Maryland, U.S. Decker, D.A., LaFleur, P.O., LeRoy, A.F. "Spontan-
eous Deposition Radiochemical Separation for Platinum Determination in
Biological Materials." Presented at the 19th Annual Meeting on Analytical
Chemistry and Nuclear Technology, October 14-16, 1975 Gatlinburg,
Tennessee, U.S.
81-A
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2.
Radioisotopes of platinum can be incorporated in the drug to
trace its distribution after administration, but the practical
considerations of isotopic abundance, decay energy, half-life,
purification of labeled preparation and cost limit such applications.
Reported analytical methods were generally developed for ore
and alloy assays; such materials are usually more concentrated and
less complicated matrices than biological materials. Of the
available analytical methods, AAS offers a relative simplicity of
equipment and lends itself to routine application to a variety of
biological specimens. Ease of application of the technique and
accuracy of the measurements depend on the tissue and concentrations
obtaining.
Equipment and Methods
Samples are atomized flamelessly at approximately 2700° in a
heated graphite atomizer (Perkin-Elmer, HGA-2000). Absorbance is
measured at 265.9 nm against a deuterium arc reference beam with a
Perkin-Elmer 303 Atomic Absorption Spectrophotometer and displayed on
an Omniscribe strip chart recorder (Model 5213-12). Aliquots of
samples requiring no pretreatment (urine, blood plasma) can be analyzed
by direct combustion in the furnace so long as the volume (typically
30 ul) introduced contains a minimum of 2 ng platinum. Tissues requiring
pretreatment are weighed, freeze-dried, and triturated to yield
homogeneous samples. Each sample is liquified and partially digested by
heating with 60% HNO, in a Teflon beaker. Digestion is completed with
addition of a mixture of 3 parts HC104 (70%) to 2 parts HN03 (60%)
(approximately 20 ml per gram of tissue). Appropriate safety measures
should be taken when using HCIO^ to oxidize organic material. When
the digest is almost dry, the residue is taken up in 0.1 M HC1,
evaporated and resolubilized in a known volume of the HC1. Aliquots
of this digest are then analyzed by comparison with working standards
prepared by dilution of a 1000 ppm hexachloroplatinic acid standard.
The method of additions has been applied successfully for analysis of
platinum in some biological materials.
82-A
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3.
The deuterium arc reference serves as a basis of correction for
broad band absorption but may be insufficient for tissues or digests
that produce profuse smoke near the atomization temperature. A modi-
fication of the HGA-2000 Temperature Controller, designed and built
at NIH, permits control of the rate of temperature rise between the
plateaus for drying, charring, and atomizing the sample. Use of the
temperature ramp generator produces a time interval between the
•appearance of the smoke peak and the platinum peak.
Results
Precision and accuracy of flameless AAS are satisfactory for most
tissues at the levels observed in the distribution studies (8 - 0.03 ppm,
wet weight basis). Immediately after drug administration in the dog,
the platinum concentration rises sharply in the kidney, liver, skin,
muscle and urine. Platinum levels in the brain remain low during the-
entire interval studied. Rapid urinary clearance of about two-hour
duration is followed by a low clearance rate measured for twelve more
days. A pharmacokinetic model is being developed based on the data
obtained.
Flameless atomic absorption spectrophotometry results obtained
using the temperature ramp generator compare favorably with preliminary
data by the more sensitive neutron activation analysis. Biological
Standard Reference Materials certified for platinum are not yet
available from the National Bureau of Standards. Their availability
may permit modification of sample preparation and the AAS procedure to
increase its sensitivity further and render it suitable for determination
of the low levels in the environment.
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Spontaneous Deposition Radiochemical Separation
for the Determination of Platinum, Palladium and Gold
in Biological and Environmental Materials
D. A. Becker, P. D. LaFleur, and A. LeRoy*
Analytical Chemistry Division
National Bureau of Standards
Washington, DC 20234
The spontaneous deposition technique has be-en used a number
of times for radiochemical separations in activation analysis
(1, 2). It has also been used as a preirradiation chemical
separation method (3). Although this technique has been used
for the analysis of one or more noble metals from inorganic
materials (2) and metals (3) , it apparently has not been
applied to the determination of noble metals in biological
and environmental materials.
The use of platinum, palladium and possibly other noble metals
in motor vehicle emission control systems has recently been made
public. It thus becomes imperative that accurate and sensitive
methods for the determination of these elements be available
for the evaluation of environmental baseline levels and any real
or imagined increases in these levels.
In addition, there is currently medical research on the use
of platinum compounds as chemotherapeutic agents in the treatment
for certain types of malignant growth. In order to adequately
evaluate the metabolic pathways and rate of discharge of these
compounds from laboratory test animals, it is necessary to have
an analytical method which is both accurate and precise.
*Guest worker at NBS; from the National Institutes of Health,
Biomedical Engineering and Instrumentation Branch, Division
of Research Services.
85-A
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The neutron activation analysis method developed and described
here is capable of quantitatively determining gold, platinum
and palladium in a wide variety of biological and environmental
matrices. The nuclear characteristics of the radioisotopes
used in this procedure are found in Table 1. As shown, the
isotope used for the determination of platinum is the gold-199
daughter from platinum-199.
In addition to the matrices described here, this method is
currently being applied also to the analysis of several new
biological standard materials currently undergoing certification,
a fresh water sediment being evaluated as a potential reference
material, and an urban air particulate material.
Experimental
All samples were prepared for analysis by lyophilization. Dry
samples weighing from twenty to two-hundred milligrams each were
placed into precleaned, polyethylene snap cap vials for neutron
irradiation. The facility used was the NBS nuclear reactor,
with irradiati.on periods of up to four hours at a thermal
neutron flux of 5.6 X 1013 n-cnr2 sec-1.
The chemical separation technique employed involved wet-ashing
the irradiated biological or environmental sample with approxi-
mately ten milliliters of a three to one mixture of concentrated
nitric and perchloric acids. This sample dissolution took
place in the presence of about one milligram each of platinum,
gold and palladium carriers, and about five milligrams of copper
holdback carrier. The samples were slowly wet-ashed on a hot
plate to fumes of perchloric acid, then taken almost to dryness.
After.cooling, they were taken.up in about three milliliters
of a three to one mixture of concentrated hydrochloric and nitric
acids until complete dissolution occurs. An additional five
86-A
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milligrams of copper holdback carrier was added at this time,
along with several milligrams of scandium and phosphate holdback
carriers. Samples were then diluted to approximately twenty
milliliters, and quantitatively transferred into appropriately
sized disposable plastic containers (e.g. centrifuge tubes,
bottles, etc.). At this point, 200 milligrams of finely
divided silver metal powder was added, and the solution-powder
mixture stirred for approximately five minutes using a magnetic
stirring bar with high agitation. The silver powder should
swirl throughout the entire solution during this period.
i
After spontaneous deposition onto the silver powder is concluded,
the solution with silver powder is quantitatively transferred
to a standard radiochemical filter chimney with a filter capable
of retaining fine particle size material. In this work, glass
fiber filter pads were used. The silver powder is washed
several times with distilled water, and great effort is taken
to ensure that all of the silver particles have oeen transferred
to the filter. At this point, the silver powder on the filter
paper is removed from the filter chimney and carefully transferred
to a plastic disposable petri dish and mounted for subsequent
counting. Separation of the silver by centrifugation was also
investigated, but filtration was found to be more effective
in removing the silver powder fines from the solution.
Aliquots of the dissolved standards are processed in exactly
the same way in order to check radiochemical recovery. In
addition, duplicate aliquots are added directly to filter paper
in the disposable petri dishes producing a geometry similar
to that of the processed sample.
Results and Discussion
Tracer studies made using this radiochemical separation
procedure showed quantitative recovery (greater than 99.5%) for
/
the gold-198, gold-199, and palladium-109.
87-A
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This analytical system has been evaluated through application
to the determination of gold, platinum, and palladium in
several available biological and environmental materials.
Results on the analysis of a bovine animal tissue (liver) gave
values of 1.8 ng/g for gold, less than 1.6 ng/g for platinum,
and less than 1 ng/g for palladium. Results for the analysis
of a powdered botanical material (leaves) gave results of 0.2
ng/g for gold, less than 9 ng/g for platinum, and less than
1 ng/g'for palladium. These analyses were made using a neutron
flux of 5.6 X 10l3 n-cm-2sec-1 for four hours, and counting
both on a 6% efficient Ge(Li) detector (gold-198, gold-199)
or LEPS System (109-palladium). For the botanical material,
the less sensitive 208 keV gamma ray peak was used due to an
interference with the 158 keV gamma ray. This interference
was apparently due to the scandium-47 daughter from neutron
irradiation of natura1ly stable calcium-47. Later determinations
using scandium holdback carrier eliminated this interference
problem.
In conclusion, the spontaneous deposition technique has been
demonstrated for the first time to be an effective radidchemical
separation method for biological and environmental samples,
and neutron activation combined with this separation technique
appears to be a very useful method for the determination of
platinum, palladium, gold, and possibly other noble metals
in biological and environmental materials.
88-A
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References
1. W. D. Kinard, D. A. Becker, and P. D. LaFleur, "The
Determination of Indium and Copper in SRM High
Purity Gold Wire by Neutron Activation Analysis,"
in Activation Analysis Section: Summary of Activi-
ties, edited by P. D. LaFleur.and D. A. Becker, NBS
TecTT. Note 548 (1970) pp. 66-69.
2. D. A. Becker, "Trace Analysis for Platinum in Glasses
by Neutron Activation," Anal. Chem. Acta 61, 1 (1972)
3. K. S. Park, R. Gijbels, and J. Hoste, "Neutron Activa-
tion Analysis of Palladium, Platinum, and Rhodium in
Lead Foam," J. Radioanal. Chem. 5, 31 (1970).
Table 1. Radioisotopes Used for the Analyses
Element
Determined
Gold
Platinum
Palladium
Radioisotope
Used
198
199
Au
Photon Energy
412 keV
Au (daughter) 158 keV (208 keV)
109
Pd
88 koV
Half-Life
2.69 d
3.14 d
13.S h
89-A
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PRESENTATION SUMMARY: ANNA M. YOAKUM
Topic: Measurement Technology and Emmission Characterization.
Sub-topic: Measurement of Pt in Biological Tissues, Urine, and Feces.
A historical presentation of the involvement of Stewart Laboratories, Inc,
in the topic under consideration will be presented. An evaluation of
methodology currently employed in autopsy tissue analysis (1) will be
discussed as it relates to future research goals.
(1) Yoakum, A. M., Stewart, P. L., and Sterrett, J. E., "Method
Development and Subsequent Survey Analysis of Biological Tissues for
Pt, Pb, and Mn Content." Env. Health Prospectives, Vol. 10, pp 85-93,
1975.
91 -A
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Methylcobalamin: Methylation of Platinum
and Demethylation with Lead
R.T. Taylor
Biomedical and Environmental Research Division
Lawrence Livermore Laboratory
University of California
Livermore, California 94550
A study was carried out to evaluate the potential for platinum,
palladium, lead, and maganese salts to be biochemically methylated.
Methylation is an important well -recognized determinant of metal toxicity,
examples being the increased hazard of methyl mercury over mercuric salts
and the apparently reduced toxicity of methyl arsenic acid relative to
arsenite and arsenate. The above metals are associated with emissions
arising from the use of automotive fuels, fuel additives, and catalytic
control devices and are therefore of special concern to the Environmental
Protection Agency. In this study methyl cobal ami n (MeB-12), a functional
derivative of vitamin B-12, was used as a biological alkylating agent.
+ Salts of Pt, Pd, and Pb and oxides of Pb all containing the metal in
a 4 valence state were observed to demethylate MeB-12 under weakly acidic
conditions. Pt(SO^)2 was unique in that it rapidly demethylated MeB-12 at
pH 7.0. Inorganic salts and oxides of manganese were unreactive at all
pHs tested (2.0-7.0); soluble Pb2+ salts, suspensions of PtOp, and KpPtC
alone were likewise inactive. Demethylation of [Me-'4C]MeB-T2 with Pb4"1"
oxides was accompanied by a proportionate volatilization of the label and
only unreacted radioactive MeB-12 was detected upon paper electrophoresis.
In contrast, acidic demethylation with KpPtClg and Pt(SO/i)2 occurs with a
significant recovery of the Me-'4C label, 75-85% and 37% respectively.
The reaction between MeB-12 and H^PtCls has been studied in more detail as
described below.
Incubation of yM levels of I^PtCls and methyl cobal ami n (MeB-12) results
in the complete conversion of MeB-12 to aquoB-12. Demethylation is optimal
at approximately pH 2.0 and is accelerated by the addition of K^PtCld. The
reaction is stoichiometric between MeB-12 and the K2?tCl6 added (1:1).
Isosbestic points at 492 nm, 367 nm, 335 nm during the course of the
reaction indicate that MeB-12 is demethylated to aquoB-12 with no
accumulation of corrinoid intermediates. Higher alkyl cobal ami ns and
93-A
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-2-
Mecobinamide react at much slower rates compared to MeB-12. Incubation of
40 yM K2PtCl6 with either 40 yM [Me-14c]MeB-12 or [Me-3H]MeB-12 followed by
lyophilization converts 70% of the label to a stable form which is associated
with Pt upon subsequent paper chromatography and electrophoresis. There is
no preferential loss of -fy relative to ^C in the reaction product. Difference
spectra indicated that the Pt reaction product had a 260 nm absorption
of [Me-'4C]MeB-12 and I
maximum. When 50 ymoles each of [Me-C]MeB-12 and I^PtCls were reacted
and subjected to Sephadex G-15 chromatography the l^c-label eluted with 260 nm
absorbing material. Further chromatography on Sephadex G-15 and CM-cellulose
yields a labeled UV-absorbing product with a 14C/Pt ratio of 0,9-1.2. Our
overall recovery was 36-42% based on the '^C. The l^C-Pt product has
absorption maxima at 260 nm and 208 nm with a minimum at 240 nm
(A24Q nm/A260 nm = 0-5)- Proton NMR spectroscopy confirmed the presence
of an H-C-Pt covalent bonding pattern (J for "H, 195pt = 78.2 Hz; T for
194pt-Me + 196pt-Me = 6.956). This Me-Pt compound is somewhat light-
sensitive; however, its moderate stability in aqueous solutions to NaCl ,
temperature, and pH suggest that it could have biological activity.
94-A
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TOXICITY OF PLATINUM (IV) SALTS FOR CELLS OF PULMONARY ORIGIN
M. D. Waters, T. 0. Vaughan, D. R. Abernethy, H. R. Garland, C. C. Cox, and
D. L. Coffin, Environmental Protection Agency, Environmental Research Center
Research Triangle Park NC 27711
The acute toxicity of tetravalent platinum was studied j_n vitro using rabbit
alveolar macrophages and human lung fibroblasts (Strain WI-38). Alveolar
macrophages obtained by saline lung lavage were exposed in tissue culture for
a period of 20 hours to platinum dioxide or platinum tetrachloride. There was
no evidence of dissolution of platinum dioxide in the test system and, although
the particles were actively phagocytized, no cellular toxicity was observed at
concentrations as high as 500 yg per ml of culture medium. Platinum tetra-
chloride, on the other hand, was soluble in the test system and after a 20-hour
exposure, resulted in loss of viability (failure to exclude trypan blue) in 50%
-4
of the cells at a concentration of approximately 3 x 10 M. Additional tests
indicated that death of macrophages was preceded by a decrease in total cellular
ATP and a reduction in phagocytic activity as compared to controls. After a 20-
hour exposure, human lung fibroblasts were rendered nonviable by platinum tetra-
chloride at concentrations comparable to those which affected rabbit alveolar
macrophages. Additional studies in the fibroblast system demonstrated that
platinum tetrachloride produced a rapid concentration-dependent decrease in
'C-thymidine uptake and incorporation into acid precipitable material. After
20 hours, these activities were 50% inhibited as compared to controls at a
platinum tetrachloride concentration of approximately 10~ M. A higher concentration
(approximately 6 x 10~ M) was required to inhibit the uptake and incorporation
of C-uridine and leucine by 50%. Studies on the ability of human lung
fibroblasts to recover following acute exposure to and removal of platinum
tetrachloride indicated the reversibility of the inhibitory effects of this
compound on DNA, RNA and protein biosynthesis.
9S-A
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Compare Pulmonary Carcinogenesis of Platinum Group Metal Compounds
and Lead Compounds in Association with Polynuclear Aromatics Using
In vitro Hamster System
Or. Philip Kane
State University of New York - Stoneybrook
The purpose of this contract is to evaluate the potential role
of platinum and lead compounds as cofactors in respiratory carcinogenesis.
The data derived from this investigation will be useful to EPA in evaluating
the hitman health hazard associated with platinum compounds emitted into the
environment in automotive exhaust. Such emissions have been suggested to be
a potential consequence of catalyst attrition in automobiles equipped with
platinum based catalytic exhaust converters. Results will aid in assessing
the potential hazards of airborne particulate lead derived from combustion
of gasoline containing lead compounds as anti-knock additives. These metals
are to be evaluated for their potential cofactor role in the induction of
neoplasia, particularly of the respiratory tract, in conjunction with the
ubiquitous environmental contaminant, and known carcinogen, benzo(a)pyrene.
The contract effort is designed to permit an assessment of the relative
hazard of platinum compounds in comparison to lead compounds as environmental
contaminants derived from automotive exhaust.
The scope of work for this contract effort entails repetitive admini-
stration by intratracheal instillation of the metal compounds under study
to Syrian Golden Hamsters. Each metal compound is to be administered alone
and in combination with benzo(a)pyrene as a saline suspension. Instillations
of benzoCa)pyrene alone, and of the combination of benzo(a)pyrene with iron
oxide (Fe203) are to be administered. The latter combination has been
extensively characterized in the past, and is expected to be a potent inducer
of pulmonary neoplasia when administered via the endotracheal route. Test
compounds are to be administered to animals once weekly for at least 16
weeks, after which animals will be maintained for their natural lifetime.
97-A
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>ross and hlstopathologic evaluation of lesions produced will provide the
indpoint of the study.
Additional studies to be conducted during the next 24 months are
mtlined in the continuation of the contract dated January, 1976. These
Include: 1) evaluation of transpulmonary absorption of lead or platinum
-------
STUDIES OF NO-EFFECT LEVEL OF Pt and Pd USING
MOUSE PULMONARY INFECTIVITY MODEL
Richard Ehrlich
IIT Research Institute
Chicago, IL 60616
SUMMARY
Mice were exposed to aerosols of platinum dioxide (Pt02) and
palladium oxide (PdO) in respirable particle size, and challenged
with airborne influenza A/2/Taiwan virus or Streptococcus sp.,
Lancefield Group C. Parameters investigated were mortality and
survival rates, pulmonary consolidation, lung edema, histopathological
changes in lungs by light and scanning electron microscopy, and
phagocytic activity of alveolar macrophages.
Mice challenged with Streptococcus at 1 or 5 hr after exposure
to Pt02 or PdO aerosol showed significantly increased mortality rates
and decreased survival time, compared to control mice challenged only
with the infectious agent. These effects were not seen when
influenza virus was used as the infectious agent. Aerosol concen-
trations to which the mice were exposed ranged from 181 to 316 mg/m3
of Pt02 and from 156 to 287 mg/m3 of PdO. This resulted in estimated
total inhaled doses ranging from 650 to 2500 yg. Atomic absorption
analyses indicated that between 75 to 90% of the metals was retained
in lungs for at least 14 days after inhalation of Pt02 or PdO aerosols
Results of limited experiments indicated that inhalation of
Pt02 aerosol, markedly diminished the ability of lungs to clear intra-
nasally instilled Staphylococcus aureus. This suggests that pulmonary
inactivation of bacteria in alveolar macrophages was significantly
altered by exposure to Pt02.
Studies supported by EPA Contract No. 68-02-1273. Presented at
Catalyst Research Program; Platinum Research Review Conference.
Rougemont, NC, December 3-5, 1975.
99-A
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":^""-y~^^i^-v«.J^ri^-\r«*.--'*
•j^t'S^^TtSS ;'~~ *• ^- f -•- LI
iU.v, DIOXIDE
100-A
-------
3
60
50
" AO
u
« 30
4J
I 20
w
u
10
1 hr
PdO »- Strep
5 hr
PdO'
•Strep
690 1100 1800
690 870 1160 1800
Inhaled Dose, yg
* p <0.05
** p <0.01
EXCESS MORTALITY AMONG MICE EXPOSED TO 'PdO AND CHALLENGE WITH STREPTOCOCCUS
1
n>
-4:
-------
o
ro
100
90
80
70
60
c
o
«H
g 50
j
40
30
20
10
0
Intratracheal
in Hamsters
PtO,
24 A8
Hours
72
Aerosol
in Mice
PtO,
PdO
14 days
RETENTION OF PLATINUM AND PALLADIUM IN LUNGS OF. ANIMALS
EXPOSED TO PtO, AND PdO
u.
£
^
3\
»*r
I
r»
(J
r '
i
-------
o
co
n
05
3
I
60
50
30
20
10
1 hr
PtO,
•Strep
*** '***
650 950 1450 1950
5 hr
PtO
650 950 1450 1950
* p^ 0.1
** p ^ 0.05
*** p .£ 0.01
Inhaled Dose,
EXCESS MORTALITY AMONG MICE EXPOSED TO Pt02 AND CHALLENGED WITH STREPTOCOCCUS
-------
TABLE 1
BACTERICIDAL ACTIVITY IN LUNGS OF MICE AFTER
INHALATION OF METAL COMPOUND AEROSOLS
BACTERIA RETAINED (%)
f COMPOUND
CONTROL
CONTROL
MID
HS_
2090
0
1935
0
GEOMETRIC MEAN
39,6
13,4
31,8
46,2
SE
8,0
2,3
4,8
5,6
.NO, LUNGS
5
4
12
6
-------
COMPARE RELATIVE TOXICITIES OF Pt COMPOUNDS AGAINST Pb
COMPOUNDS USING IN VITRO MACROPHAGE SYSTEM
Dr. Donald Gardner
Environmental Protection Agency
Health Effects Research Laboratory
(Summary of work performed by Dr. Eula Bingham, University of Cincinnati)
This study compared and evaluated the cellular response of the lung,
especially the pulmonary alveolar macrophage, to the effect of inhalation
of Pt and other metal compounds using the alveolar macrophage (AM) as a
biological indicator of toxic effects. In this study young rats were
exposed to an aerosol (150 ug/m ) of either PtCl, NiCl, NiO, Pb-O.,,
PbCl or CdO for 6 hours/day, 5 days/week. At various time intervals, the
animals were removed from the chamber, sacrificed and the lungs lavaged
so as to isolate the pulmonary cells (AM). The amount of metal deposited
in the lung was determined by atomic absorption technique. Some of these
substances that were tested (CdO, NiCl2> PdCl, and PtCl) did not statis-
tically alter the number of lung cells while NiO caused an increase and
Pb203 caused a depression in number of lavable pulmonary cells. This
indicates that one cannot assume that the inhalation of particles per se
induces a non-specific increase in pulmonary cells. In these experiments
the number of isolated cells were dependent on the specific metallic parti-
cle inhaled. The reaction of the lung to these particulates may depend
upon a number of factors such as solubility, physical properties, on
specific metals. Also determined in these studies were the enzymatic activity
i.e. acid phosphatase, alkaline phosphatase, and lysozyme in alveolar macro-
phages exposed to platinum. Preliminary analysis indicates an increase in
cellular lysozyme and acid phosphatase and no apparent effect on alkaline
phosphatase.
/
-------
Dr. James H. Taubler
St. Vincent's College
Allergic Induction in Experimental
Animals Using PtSO^ and PdSO^
Various routes and metal concentrations were used to induce
an allergic state in rabbits, guinea pigs and mice. Subcutaneous
and intravenous injections of 0.3, 0.1 and 0.05 mg/ml of platinum
or palladium sulfate three times per week for four weeks failed
to induce either immediate or delayed allergy as determined by
skin test in rabbits and guinea pigs or by footpad test in mice.
Skin exposure to 0.25g and O.lg paste applied once a week for five
weeks also failed to induce the allergic state. Intravenous
injection of a palladium-albumin complex failed to induce allergy
also. However, subcutaneous injection of the palladium-albumin
complex induced a delayed type allergy in guinea pigs. Passive
transfer via spleen cells was effected.
Additionally, the blood, urine and splenic tissues were
assayed by Atomic Absorption Spectrophotometry for platinum and
palladium. When the metals were injected intravenously no trace
of the metals was found. In rabbits and guinea pigs subcutaneously
injected with platinum, no metal was found in serum or urine.
However, in a similar experiment using palladium, guinea pigs
showed significant levels in the urine. Regarding splenic uptake
of the metal, only platinum was found in the spleens of rabbits,
guinea pigs and mice subcutaneously injected with PtSO^.
To date the following conclusions can be made:
1) There is no allergic induction in rabbits, guinea pigs or
mice injected subcutaneously or intravenously with platinum or
palladium sulfate.
107-A
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2) In animals injected intravenously with PtSOjL or PdSO^, no
levels of metals can be found in the serum, urine or spleen.
3) In animals injected subcutaneously with PtSO^ or PdSO/^ only
palladium enters urine of rabbits and guinea pigs.
*O In guinea pigs subcutaneously injected with a palladium-
albumin complex, a delayed type allergy develops which can be
passively transferred via lymphoid cells.
108-A
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THE EFFECTS OF PLATINUM SULFATE ON LEUCOCYTE AND PLATELET METABOLISM
AND ON IMMUNOLOGIC RESPONSES IN THE RABBIT
Kenneth D. Lunan, Ph.D.
Senior Biochemist and Program Manager
Environmental Biochemistry Program
Biomedical Research Department
and
Ted A. Jorgenson, M.B.A.
Toxicologist
Toxicology Department
Life Sciences Division
Stanford Research Institute
This research is a comprehensive study of biochemical, physiologi-
cal, immunological, and mutagenic effects induced in blood leucocytes,
blood platelets, and major organs of the mammalian body by exposure to
platinum sulfate. This material may be a major attrition product of
catalytic converters, and large segments of the population may be
exposed to it in the future. In addition, this research may serve to
establish the feasibility of using formed elements of the blood, an
easily accessible body tissue, as monitors of the body burden of heavy
metals.
Rabbits were injected intraperitoneally daily with 2, 15, or 20
mg/kg of Pt(804)2 f°r 1 or 3 weeks and then sacrificed. Others were
allowed to recover for 1 week, 1 month, or 3 months before sacrifice.
Control animals treated with saline were included. Leucocytes and
platelets were isolated and examined by numerous criteria for effects
due to Pt(SO4)2 treatment. The ip LD50 for Pt(SO4)2 in rabbits was
found to be 210 mg/kg, with a range of 134 to 322 at the top 95% con-
fidence limits.
109-A
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Skin and eye irritation studies in rabbits and skin sensitization
studies in guinea -pigs were also performed. Pt(SO4)2 was applied at a
concentration of 0.5 g/sq in. to abraded and unabraded skin patches
for up to 72 hours. No effect of Pt(S04)2 was noted. Guinea pigs
were injected subdermally with a 0.1% solution of Pt(SO4)2 on alter-
nate days for 20 days. Two weeks after the last injection, an addi-
tional challenge injection was given. Examination of the injection
site 24 hours after each injection indicated that Pt(S04)2 did not
cause skin sensitization in the guinea pig- Eye irritation studies
were performed by instilling 0.1 g of Pt(SO4)2 into the conjunctival
sac of the eye of the rabbit. The material was then washed out after
30 seconds or 5 minutes. In other rabbits, the material was not
washed out. The cornea, iris, and conjunctiva were examined after
1 and 4 hours and then daily for 14 days. The rabbits whose eyes were
washed 30 seconds after instillation of the test material showed a
transitory effect and were generally normal after 9 days. However,
the eyes of rabbits in the 5-minute wash and no-wash groups were more
severely damaged and had not returned to normal by 14 days. In many
animals, the amount of conjunctival swelling was so great that either
the eye surface could not be observed at all or only a small portion
of the cornea and iris could be viewed for scoring. Thus, Pt(S04)2
should be regarded as a moderate to severe eye irritant, depending on
wash conditions.
In the leucocyte experiments, the majority of the effects were
observed in the exposed animals from two treatment groups—the one
110-A
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subjected to 3 weeks of treatment with Pt(SO4)2 at 20 mg/kg and the
one allowed 1 month to recover after 3 weeks of treatment with Pt(SO4)2
at 15 mg/kg. In the former, blood values were not affected. RNA syn-
thesis was depressed (DNA synthesis was depressed after 1 week of
treatment). Also, protein turnover after 24 hours was retarded. In
the latter group, only one labeling parameter was affected—the phos-
pholipid turnover, which was markedly retarded after 24 hours. The
other values affected in this experiment were blood parameters. The
mean corpuscular hemoglobin concentration and the lymphocyte count
were low, whereas the eosinophile count was markedly high. In general,
in vivo Pt(SO4)2 treatment had only modest effects on leucocyte
metabolism.
Several preliminary iji vitro exposure experiments indicate that a
2-hour exposure to 10~4 M Pt(SO4)2 caused a 50% depression of DNA
synthesis.
Numerous alterations of platelet metabolism were observed in all
experiments involving animals exposed to 15 and 20-mg/kg of Pt(SO )2.
5-Hydroxytryptamine uptake was stimulated by 1 and 3 weeks of Pt(S04)2
treatment. The stimulation remained after 1 week of recovery but was
gone after 1 month of recovery. 0-Glucuronidase activity was elevated
in the platelets from these two treatment groups and had returned to
normal after 1 week of recovery. Aryl sulfatase was elevated in the
animals treated 1 week and after 1 week and 1 month of recovery.
Finally, phospholipid synthesis was depressed after 1 week of treat-
ment, stimulated after 3 weeks of treatment, and again depressed after
1 month and 3 months of recovery.
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Several whole blood values were affected in the 1-week, 1-month,
and 3-month recovery experiments. Generally red blood cell and plate-
let counts were elevated, and hemoglobin and hematocrits were low.
Also, the volume, hemoglobin content, and hemoglobin concentration of
the average red blood cells were low. Thus, Pt(S04)2 produced a func-
tional anemia that did not appear until 1 week after treatment had
ceased and that persisted for at least 3 months-.
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Cytogenetic Analyses of the Effects of Platinum
Ann D. Mitchell
Biochemical Cytogenetics Program
Biomedical Research Department
Stanford Research Institute
Menlo Park, California 94025
Cytogenetic analysis is one of the most widely used methods for
assessing the cellular damage caused by environmental pollutants.
The examination of chromosomal aberrations in cells after in vivo
exposure to environmental pollutants can reflect both the direct
action of mutagenic agents and the indirect effects of an altered
metabolism. The observation of Cytogenetic damage occurring in a
controlled in vitro environment can be of value for determining the
mechanisms of possible mutagenic action.
This study was proposed because: A search of available litera-
ture had failed to reveal any previous testing of platinum salts for
mutagenic activity, although certain other metals had been shown to
induce chromosome breaks and aberrations and/or mutations; some
researchers indicated that certain platinum complexes having a
marked antitumor activity may induce a primary lesion in DMA through
the formation of an intrastrand purine dimer; and some investigators
suggested that metal ions may enhance the aberration frequency of
clastogenic agents by complexing with the broken chromosomes,
thereby preventing the repair of the breaks.
We are conducting the following seven experiments to determine
whether platinum induces chromosomal aberrations or prevents the
repair of Cytogenetic damage induced by a known mutagenic agent,
ethylmethanesulfonate (EMS): (l) Examination of chromosomes from
rabbit leucocytes exposed in vitro to Pt(SO4)2, with one-half of the
samples also exposed to EMS; (2) a similar ±t\ vitro experiment using
human leucocytes; (3) examination of chromosomes from leucocytes of
rabbits exposed ii\ vivo to Pt(SO4)2 for 1 and 3 weeks, with EMS
added ±r\ vitro to one-half of the samples; (4) a similar experiment
using leucocytes from rabbits recovering for various lengths of time
from a high dosage of Pt(SO4)2; (5) examination of chromosomes from
bone marrow cells of rats exposed to acute and chronic dosages of
Pt(SO4)2; (6) chromosome banding analysis of the more significant
samples from the previous studies; and (?) use of an assay system
developed at SRI to evaluate the effects of Pt(SO4)2 in inducing
and/or inhibiting repair of DNA damage.
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Toxicities of Compounds of Platinum and Palladium
as Measured by Biochemical Parameters
David J. Holbrook, Jr.
University of North Carolina
Studies have been conducted on the effect of the dietary administration
of salts of lead, manganese, platinum and palladium on the following:
the growth rate of male rats, the organ weight of five tissues (liver,
kidney, spleen, heart and testis), and the tissue content of DNA, RNA and
protein. In general, dietary levels of PbCl2, PdCl2, PdO and PdSO^ greater
than 10 mmoles/kg feed were necessary to restrict the weight gain of the
rats. Soluble salts of Pt*+, namely PtCl4 and Pt(804)2-4H20, at levels of
approximately 2 mmoles/liter drinking fluid, were sufficient to restrict
the weight gain of treated rats. Dietary PbCl2 markedly increased the
size of kidneys in treated rats. In almost all other studies, however,
the dietary administration of salts of lead, manganese, platinum or
palladium did not markedly or consisently alter the organ weights of the
metal-treated rats. The dietary administration of PbCl2, PtCl4 or Pt(S04>2
4H20 for 4 weeks did not alter the content of DNA, RNA or protein in
liver, kidney or spleen (when the content is expressed per gram of wet
tissue).
The intraperitoneal administration of PtCl^ or Pd(N03>2 at levels
of 28 or 56 ymoles/kg body weight decreased the thymidine incorporation
into DNA of spleen, liver, kidney and testis. Spleen was most sensitive
to both the platinum and the palladium salt. In liver, DNA syntheses in
parenchymal cells and stromal cells were about equally sensitive to PtCl4.
The administration of PtCl4 or Pd(N03)2 did not decrease the entrance of
the labeled precursor into the tissue, and PtCl4 did not alter the relative
amounts of thymidine and its phosphates in liver or spleen.
The effects of various salts of platinum or palladium were determined
on the parameters of the microsomal mixed-function oxidase system from
rat liver. The intraperitoneal injection of PtCl^ or Pd(N03)2 at 56
umoles/kg, Increased the hexobarbital-induced sleeping time in vivo and
generally decreased the aminopyrine demethylase in vitro and the microsomal
content of cytochrome P-450. The dietary administration of various salts
of Ft or Pd for one week generally decreased or had no effect on the para-
meters of drug metabolism by isolated microsomes and after 4 or more weeks
generally had no effect on, or increased, the parameters. The addition of
0.15-0.2 mM PtCl4 or 0.2-0.3 mM Pd(N03)2 to the incubation medium (containing
5 mM MgCl2> inhibited the aminopyrine demethylase of isolated hepatic
microsomes by approximately 50%.
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Studies of Platinum and Palladium in Human Tissue
Using Selected Populations Including Occupationally
Exposed Individuals
Mr. John Prevost and Dr. Donald Johnson
Mr. Prevost will present information on the selection of sites
for the baseline and occupationally exposed populations, on the
recruitment of study participants and on the description of the study
participants. Dr. Johnson will present a summary of the results for
Pt, Pd and Pb in these populations.
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Evaluation of the Mutagenio Potentials of Platinum Compounds
Shahbeg Sandhu
Department of Biology
N. C. Central University
Durham, North Carolina
This study was initiated to screen a number of platinum com-
pounds for their possible mutagenic effects. An established cell
line, L5178Y mouse lymphoma cells, is employed as a test system.
These cells are known to be diploid at thymidine fcinase (TK)
locus. The selective media for isolating TK competent cells
(TK •••/- and TK -:/') and TK deficient cells (TK -/-) has also
been developed. The compound under scrutiny is considered to
be mutagenic if it has the ability to change the TK competent
cells to TK deficient cells and virj? vprsa^ with frequency com-
parable to that of the known mutagens.
In addition to the jai vitro testing, presoaked and dry seeds
of PhaspoluR atirnalja are exposed to different concentrations of
platinum compounds. The data is recorded on germination, charact
teristics, growth and various morphological characters in M^, Mg
and Mg generations. Currently the following compounds are under
study in our laboratory:
Platinum diammino dichloride, Ammonium chloroplatinite,
Platinum dinitrodichloride (cis and trans isomers), Ammonium
chloroplatinite, and Platinum chloride.
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DEVELOPMENT OF SENSITIVE BIOCHEMICAL AND
BEHAVIORAL INDICATORS OF TRACE SUBSTANCE EXPOSURE
Dr. Edward J. Massaro
Department of Biochemistry
State University of New York at Buffalo
Buffalo, New York 14214
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In adult male mice (Swiss-Webster CFW strain: Carsworth Farms, New
York, New York), the tissue/organ distribution and effects of platinum on:
(i) levels of selected neurotransmitters; (ii) energy production through
glycolysis and; (iii) rates of DNA, RNA and protein synthesis are being
investigated. Simultaneously, and coordinated with these studies, behav-
ioral analysis, including open-field, passive avoidance, maternal and social
behavior are being undertaken.
Three routes of administration are being investigated: intragastric -
because of its possible environmental relevance; and intravenous and sub-
cutaneous - for monitoring the effects of rapid and slow sytemic distri-
bution, respectively. The latter two routes also will avoid confounding due
to possible binding of the compound to food materials; microfloral inter-
actions; etc. inherent in the IG route of administration. Seven day LD,-0
data are being established to serve as toxicity parameters.
Originally, Ft as Na_PtCl, was being investigated. LDc0 levels for
this compound have only been estimated (90% confidence intervals by probit
analysis). However, ultra-accurately defined toxicity levels are not nec-
essary for the rough definition of experimental parameters and pilot studies
of open-field behavioral effects and tissue/organ distribution at the ap-
proximate LD1f. and LD levels have been initiated. Reasonable estimations
of the LD,_ and LD-j. levels for each compound for each route of administra-
tion were made by simple linear regression of % responding on dose.
The open-field behavior (ambulations, rearings) of the animals was ob-
served at four time periods postadministration (4 hours, 1 day, 3 days and
seven days). Tissue/organ distribution of Pt is undergoing analysis.
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Ten animals were assigned to each treatment group. Injection solu-
tions were prepared from a 50 ppm Pt stock solution, which was adjusted
to pH 7.4 with NaOH, and diluted with phosphate buffered saline (PBS:
0.14 M NaCl, 0.01 M NaPO,, pH 7.4) to the appropriate concentrations.
For purposes of simplification, each time group was run in a block starting
with the four hour groups and preceding in order to the seven day groups.
When the three day and seven day groups were observed; inordinately high
death rates were encountered compared with the originally estimated LD
levels. Concievably, the LDin and LD_,. dose levels could have been mis-
calculated; but no miscalculations were discovered. Nevertheless, the LD
level was reestimated taking into account the new lethality data and another
open-field study was undertaken employing the reestimated LD9t-' Again,
there were excessive deaths at 3 and 7 days. Rechecking each step of the
methodology revealed that the pH of the injection solutions had drifted
with time from 7.4 to approximately 4.0. This down-shift was confirmed in
a separate experiment. The drift probably results from a slow hydrolysis
2- 2-
of the PtCl, to PtC1..0H liberating HC1. Ex post facto, it was learned
0 J
that such an hydrolysis effect occurs with other inorganic chlorides. Al-
though we have uncovered no information in the literature on this point,
it seems reasonable to assume that pH drift was responsible for the time-
dependent alterations in toxicity of our injecting solution.
In view of this technical difficulty, interpretations of the results
of the pilot open-field study are confounded; but the data were subjected
to analyses of variance.
Four behavioral response variables were measured during the 5 minute
trial in the open field: ambulations (automatically recorded); rearings;
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-3-
initial latency (in the six inch center circle); and the number of recros-
sings of the center circle during the trial. The analysis was, therefore,
of a 3 (route) by 4 (dose) by 4 (time) multivariate design and The Multi-
variance (J.D. Finn, 1972)* computer program was used to perform the analysis.
Since certain cells had less than 10 animals due to deaths, the design was
nonorthogonal which precludes exact significance tests of main effects in
the presence of significant interactions. Similar two way Anovas also were
conducted for each time since the experimental conditions were more homo-
geneous within each time.
From the overall analyses, most of the possible significant** effects
were seen in the ambulations and rearings variables. For the route effects,
IG had the highest scores across variables; IV had the lowest; and SC was
intermediate. For the dose effect, ambulations and rearings were depressed
at all dose levels compared to saline controls. The time effect was not
"significant". The route by dose interaction was the only "significant"
interaction. In the two way analyses at each time, the dose effect was
"significant" throughout while the route effect was "significant" only at
the first three times and diminished with time as would be expected. The
route by time interaction also was "significant" only at the first three
times.
In spite of the experimental and statistical ambiguity of this pilot
study, we believe the results and our own observations of the animals
strongly suggest that Na.PtCl, has an effect on behavior which would be
characterized as depressant. The effect appears most strongly shortly
*Finn, J.D. (1972) "Multivariance", National Educational Resources,
Inc., Ann Arbor, Michigan,
**Not certainly significant because of the experimental and analytic
difficulties mentioned above.
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-4-
after administration and diminishes with time. The effect also depends
on route of administration: IV, the route of most rapid uptake, exhibits
the greatest effects; IG, the route of slowest uptake, exhibits the smallest
effects.
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APPENDIX II
Platinum Chemistry Pertinent to the Catalyst Program
During general discussion at the conference, the need was expressed
for a short summary of useful information on platinum chemistry. Rather
than provide the requested information separately to the investigators
who expressed interest, it is hereby presented for general use. Questions
should be directed to the author, Dr. Richard Thompson, Environmental
Monitoring and Support Laboratory, Environmental Protection Agency, MD-78,
Research Triangle Park, North Carolina 27711.
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Appendix IIA
Platinum Chemistry Pertinent to the Catalyst Program
The catalyst program must necessarily be concerned in part with the
transition metals, since one of their characteristic properties (not
shared with representative metals) 1s catalytic activity. The use of
platinum palladium and other transition metals 1s based primarily on
catalytic activity, an empirical property. Because of the known chemical
nature of these elements and their compounds, we may make estimations of
expected chemical behavior in the environment with some confidence in
most cases. It 1s of Interest that prior to the use of platinum as a
catalyst in automobiles the environmental levels 1n matrices such as
atmospheric particulate matter were so low as to defy detection by all
methods attempted other than Isotope dilution mass spectrometry, even
when a kilogram sample was taken.
Elemental platinum is Inert to simple acids, but will react with
aqua regia (3:1 concentrated HC1:HNO,), chlorine, bromine, sulfur and
fused alkali and cyanide. None of these substances exist naturally.
The oxides are not thermally stable, decomposing below 300°C. (A temper-
ature of about 650°C is attained in automotive catalytic converters).
Biological activity is not expected to result in formation of compounds
from elemental platinum.
One of the notable properties of the transition metals 1s that they
form many and stable coordination complexes. These can be visualized as
being formed by the coordination of Ions or molecules with metal Ions.
The mechanism of this coordination is the donation of an electron pair
by tne ligand (1on or molecule) to the bare metal ion. As an illustration,
consider "Pt(SO,)V which 1s said to be deliquescent. This can be
+ B
pictured as a [Pt(H00)g]4 species accompanied by two SO^ . The hexa-
coordinated platinum ion can be shown 1n crystalline species to be
octahedral, and all six of the Ugand positions are equivalent. If one
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dissolves Pt(SO.)2 in water and allows the solution to stand, In time,
the [Pt(H20)6]4 Ion will yield Pt(OH)4 ' nH20 + 4H* which can be expressed
as Pt02 * nH20; heating the solution will accelerate the process.
In the catalyst program, we are Interested In the environmental
changes that will ensue from the introduction of platinum and its compounds
(and other catalyst metals) into the environment. Unfortunately, the
exact nature of the species emitted from a converter 1s not knowny but
Is most likely to be the element, or oxides and halides. These can be
used in inhalation studies, but for other in vivo experiments a soluble
form is desirable.
At the Quail Roost Conference, it became painfully obvious that
commercial samples of "platinum sulfate" and "platinum chloride" were
most likely hydrolytic end products resulting from the reaction of these
deliquescent substances with water from the air.
A possible approach for in vivo studies could utilize fresh solutions
of M2PtCl6 (where H is K*, Na+, NH4* etc.) The (PtClg)* 1on 1s stable
1n cold water for some time and the addition of M Cl (if not objectionable)
will further stabilize the complex In solution by forming a pool of
chloride Ions to compete with water (and Its constituent ions) as a
Ugand. Furthermore, the ion Is colored, and absorbs light very effec-
tively 1n the near ultraviolet. From the literature :
[PtCl6]"
Wave length (nm) e 6" 6
453
353
262 24,540 2,250 2,300
where e is the molar absorptivity and 6~ and 6 are the half-band widths
towards Increasing and decreasing values.
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50
790
24,540
1,900
1,700
2,250
-------
Therefore absorbance 9 (cone) (path length) (absorptivity) and 1n a
1 cm cell, a 4 x 10 M solution of hPtCl would have an absorbance of
0.1 at 262 run. One could thus measure the absorption at 262 ran prior to
administration to test animals to ascertain whether or not the solution
1s *
1. C. K. Jorgensen, Acta Ch1m. Scand. JjD, 518 (1956).
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Appendix IIB
Spectrophotometric Verification of PtC1g~
1. Obtain a supply of sodium hexachloroplatinate(IV) (Na^PtClg) or
chloroplatlnic acid (H^PtClg). Standard solutions typically used
for calibration of instruments are available. Be certain from the
supplier (got a guarantee) that the contents are the chemical form
cited above. [A SPEX Industries standard available in our labora-
tory was suitable.] Such solutions typically contain 5000 yg Pt/ml .
2. Obtain clean glassware of the proper si^e. Make up only as much
solution as will be used within 4 hours or solution preparation.
These solutions will deteriorate. Therefore do not store them.
If for exerr.ple one liter of final solution were desired, the standard
were 5000 ug/ml and the final solution concentration desired v/as
5 yg/ml a one nil pipette and a one liter volumetric flask are needed.
3. Prepare the solvent ty be used for diluting the standard. (It is
assumed here that -isotonic saline will be used.) Do not use plain
water.
4. Prepare solution. Pipette the correct number of ml of standard
solution into the volumetric flask and dilute to volume with isotonic
sarine. Mix by inverting the flask about 10 times.
5. I.ocate0a spectrophotometer capable of measuring absorbance at 262 nn
(2620 A) such as a Gary Model 14 or a Beckman DLL Th? 262 nm wave-
length is below the range obtainable on inexpensive colorimeters
such as the Spectronics , Co lemons, and Turners.
Instructions which follow are for a Gary 14.
6. While "power" switch is off, install a 0-1 absorbance slide-wire
cartridge being careful not to scratch the wire spindle.
7. Turn on coolant watr.-r supply to UV lamp before tiTnin-j on instrument
"power" switch and UV lamp switch. Allow lainp and irisvrunieni tc
warm-up for 1/2 hour.
8. Set "Source" ar.d '"Detector", selectors to "UV," "IR det" to "out,"
and pull-out phototube shutter.
9. Set wavelength to 262 nm (2620 ft) with dial approaching 262 from
higher numbers.
10. Fill two 1-cm (10 run) optical cuvette cells about 3/4 full with
same solvent to be used for diluting Pt standard. T?p cells to
dislodge any air hubbies, and v.-'ipe optical surfaces clean with
soft tissue.
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11. With "master" switch off (but "power" switch on), position the two
optical cells (both containing solvent) in the two optical beams
such that light will pass through the unfrosted sides of the
cuvettes.
12. With lids on the optic chambers, turn on "master" control and with
"slit" program and "pen" systems in "on" position set pen at 0.000
absorbance 'level on chart recorder using "balance" dial.
13. If pen movement is more than 0.005 absorb?r,ce, pen dampening can
be set higher (on Gary 14 the dampening control knob is located
on back panel behind the swing-out recorder).
14. With "master" switch off ("power" switch left on), remove cell from
"sample" optical chamber, rinse out cell repeatedly with deionizcd
water and finally with acetone to dry it.
15. Pour 5 ug/nil Pt solution into clean, dry cuvette cell (used earlier
with solvent to set zero absorbance). Tap cuvette to dislodge any
visible air bubbles, and wipe outer optical surfaces clean with
soft tissue.
16. Place cuvette with Pt solution in "sample" optical chamber and
close chamber tightly with lid.
17. Turn "master" switch on and record position of recorder pen to
determine absorbance value.
18,
Observe pen position as wavelength dial is turned slowly in both
directions (50 rim) to locate maximum absorbance value.
19. If maximum absorbance is located at different wavelength setting
than used in step £14 above, rinse "sample" cuvette repeatedly
in dc.-ionized >..'nter and refill with solvent (after drying cell
with acetone) to verify that ebsorbance v.ith solvent in both
cells reads 0.000 at this other wavelength setting. If not 0.000
absorbance add or subtract reading (as appropriate) to maximum
found in #15.
20. UV scan for 5 yg/ml Pt in water (from Spex Indust. Std.) is
attached. NaCl (0.9%) or HC1 (2N) did not alter this scan when
measurements were conducted within 1/2 hour after preparation.
The absorbance at 2400 fl and at 2850 fl should both be very nearly
equal and be equal to 1/2 of the maximum absorbance which should
occur at 2620 .
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21. The expected absorbance at 2620 $ for an expected concentration should
scale linearly as follows for a 1 cm path length cell.
Cone. Absorbance
1.0 yg/ml 0.125
2.5 yg/ml 0.313
5.0 pg/'ral 0.625
7.5 i:g/inl .938
Values shoi'ld be within about 15%'of the above. 1 cm path length cells
are standard. However, if path lengths differ values for absorbance
scale linearly, [e.g., for a 1/2 cm path length cell 8 ug/ml solutions
have an absorbsnce of 0.50. A 2.5 y.g/ml solution in a 2 cm cell should
have an absorbance of .625.]
22. If solutions more concentrated than 7.5 yg/ml are to be utilized
prepare then at the desired concentration. Take a small sample, of
known vclurr.e e.rd dilute it in a volumetric flask with normal saline
so that the f'iral concentration is less than 7.5 yg/ml. Calculate
the conceivLration expected in this diluted sample as
sample) = conc> Qf di]uted ^
or
V W I U i: I *i- V I
Examine this diluted sample in the spectrophotometer. Absorbance
should cicree with that expected for the calculated concentration.
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— '1
0-7
o.c
.- 1
t -
0.0
•o./.
0.
\.."-"•;
.._ .1 j
!_.
Cj
r»
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TECHNICAL REPORT DATA
(/'lease read /nUnictioris on the reverse before completing)
I. REPORT NO.
4. TITLE AND SUBTITLE
3. RECIPIENT'S ACCESSION>NO.
Proceedings - Platinum Research Review Conference
5. REPORT DATE
April 1976
6. PERFORMING ORGANIZATION CODE
7. AUTHORIS)
8. PERFORMING ORGANIZATION REPORT NO.
p PERFORMING ORGANIZATION NAME AND ADDRESS
Catalyst Research Program
Criteria and Special Studies Office
Health Effects Research Laboratory
Research Triangle Park, N.C. 27711
10. PROGRAM ELEMENT NO.
1AA601
1 1. CONTRACT/GRANT NO.
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
This is an unpublished report.
16. ABSTRACT
The Catalyst Research Program was initiated in FY 1974 by the EPA
Administrator, Russell E. Train, concurrent with the Agency's Nov. 1973 decision
to permit the use of noble metal oxidation catalyst as an emission control device
on 1975 model year passenger vehicles. Early research suggested that slight
emissions of platinum and palladium might be emitted.
The CRP's objective as it pertains to the noble metals, then, was to develop
an information base on noble metals, particularly platinum, in many areas.
The areas of study for the CRP's platinum research effort include: emissions
characterization, measurement methodology development, bioenvironmental impact,
health effects assessment, body burden tissue analysis, and epidemiology.
After the platinum research effort had been under way
apparent that the data being reported by researchers should
appropriate forum for informative exchange and discussion.
The conference presentations, areas for further study
emphasis or direction of current study, and priorities for
are briefly discussed in this report.
for a year, it became
be discussed at an
and possible changes in
these recommendations
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
catalytic converters
precious metals
platinum
palladium
h.IDENTIFIERS/OPEN ENDED TERMS
c. COSATI Hclil/Cioup
06 B
13. DISTRIBUTION STATEMENT
UNPUBLISHED
19. SECURITY CLASS fflm Heporl)
UNCLASSIFIED
21. NO. OF PAGES
154
20. SECURITY CLASS (This page)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
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