Document dO5Vrn9xgmJLGBKQE9zo123e
FOR DU PONT USE ONLY
AR226-2923
cc;
Haskell
E. I. du Pont de Nemours and Co., Inc.
Laboratory for Toxicology and Industrial Elkton Road, P. 0. Box 50,
Newark, Delaware 19714
Medicine
HASKELL LA30RATORY REPORT NO. 110-85
MR NO.
Material Tested
Haskell Nos.
SUMMARY
UlB BiBIHBHHIIIlHHHr'^' Grouo.s.o.^iale Cri :CD(SD)BR rats
ofeitherflBBBp(an aqueous ^usDen^lon_
or fH^sol i d
Different particle size^nstnbil^onsweregenerated
to aerosol atmospheres
a single 4-nour perTod.
to determine the effect
of particle size on the toxicity of these materials. Further, the relation
ship between expected pulmonary deposition (based on particle size) and
mortality was investigated.
)U_J^BRand|U For bo
the ALC increased with
^BUff^9 size. For |
A.l.-C^_inncc.ieased from 42 mg/m at
mg/m at 6..1V um MMD;
^^------theALC increased from
Foll7r^^~l to 360 ing/inn*" at 5.6 um MMlT.
increasing particle
1.6 um MMD to 170 24 mg/m at 1.7 um MMD
For these materials, the fraction of the total test atmosphere expected to deposit in the alveolar I'egion (particles smaller than 3.1 um) was most
closely associated wLti^^^^ity. However, this relationship was
unequivocal only for|PBim|xposures; one exposure to a low concentration
of----krtides smaller than 3.1 um caused deaths which can not be
expTa^edby expected pulmonary deposition. The atmospheric concentration of
total respirable aerosol did not show a clear dose-response.
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HLR 110-85 Page 1 of 14
INTRODUCTION
vas extremely toxic by inhalation when tested FALCof 42 ing/in"; HLR-423-83). The purpose of
.^^--L^f------^^------P ^^^^^^IBBKl ^tI.DhHi1xsSicsbitLtyuUodUyyf--wi--adsii --t.o.^--d_e_--t._e.r--.m...^i-n.H_e^t^.h.---e_
--e.-f.--f_e_c--_tjBo.
f
T
pa
wo
rticle
forms
siZfi^n^heTnhalation
'fmlmBBRfftifteferereieiess^^^^^^^^^^w^aa^^^^f^flffflWl^^eeoius suspension|H------Bandonewas
HH| ^'Thheeppuurrccmniaatteenn'al
For both materials, ApproximateTetnai
Concentrations (ALC^s) wer e determined for various particle size atmospheres.
The ALC was defined as the lowest atmospheric concentration tested which
caused the death of 1 or more rats either on the day of exposure or within 14
days post exposure. Further, the relationship between expected pulmonary
deposition (based on particle size) and mortality was investigated.
MATERIALS AND METHODS
A. Animal Husbandry
Young adult male Cri:CD(SD)BR rats were received from Charles River Breeding Laboratories, Kingston, New York. Each rat was assigned
a um'qi/e 5-digit identification number which corresponded to a numbered card affixed to the cage. Rats' tails and cage cards were color-coded with water-insoluble markers so that rats could be identified after
exposure. Rats were housed singly in 5" x 11" x 7" suspended, steel-mesh cages in rooms targeted to have temperatures of 25 ^ 2C and
50 ^_ 10'i relative humidities on timer-controlled 12 hour/12 hour light/dark cycles. Rats ware quarantined for one week prior to testing, and were weighed and observed twice during the quarantine period. Except dun no exposure, Purina Certified Rodent Chow #5002 and water were available ad libitum.
B. Exposure Protocol
Groups of 6 rats, 8 to 9 weeks old and weighing between 224 and 297 grams, were restrained in perforated, stainless steel cylinders with
either^H^UUfp'n conical nose pieces. Each group was exposed nose-flii^^oj^^^iflal e,
4-hour period to an aerosol atmosphere of
air.
Rats were weighed prior to exposure and observed TOF clinical sTgns
during exposure. Surviving rats were weighed and observed daily for 14
days post exposure, weekends excluded except when deemed necessary by
the rats' condition.
Gompany SanffTzed. Does not copfa.n TSCA CBI
Sge^o^M
C. Test Material
2. TBCU (H-15.219)
Physical Form:
Purity:
Contaminants:
Synonyms:
;hemica1s and Pigments Department Jackson Laboratory
Other Codes:
Stability:
Submitted by:
^"he test material was assumed to be stable
throughout the exposure phase of the study. Based on the supplier's specifications, the test material was stable at the temperatures
nemicals and Pigments Department Jackson Laboratory
D. Atmospheric Generation
ofUHBpre Aemsi^^^BOSpheres
generated by pumping
a Spraying Systems nebulizer. Air introduced
^l^dlimr^0 at thenebuTTzer aerosol ized th:.' test material, and swept the
aerosol stream through a cyclone elutriator and into the exposure
chamber. Particle size distribution'; were shifted toward larger
particles by removing the cyclone, using different sized nebulizers
CosyipsrtVSanr^ei-i D"-^ no, r,^^ TftrA f^m
HLR 110-85 Page 3 of 14
and changing the airflow. During one exposure, the nebulizer and the cyclone were heated to approximately 100C,
2. tfUklhl5^219!
offBrcre Aerosol atmospheres
generated by pumping melted
test material into a Spraying^ystems nebulizer. The test material was heated to 60-132C during generation. Air introduced at the
nebulizer aerosolized the test material, and swept the aerosol
stream into the exposure chamber. The air was preheated in a furnace heated to 120-278C during generation. Particle size
distribution was shifted toward larger particles by changing the
test material and air temperatures, airflow and nebulizer size.
E. Analytical
The atmospheric concentration of particulate was determined at approximately 15- to 30-minute intervals by drawing known volumes of
chamber atmosphere through pre-weighed, glass fiber filters. Filters
wera weighed on a Cahn Model 26 Automatic Electrobalance. Atmospheric
concentration of particulate was determined from the filter weight
differential before and after sampling.
During each exposure, the particle size distribution was determined with a Sierra cascade impactor. In addition, for each exposure, the estimated atmospheric concentrations of particles smaller than 3.1 and 13 urn were calculated from the total atmospheric concentration and particle size data. Chamber temperature was monitored with a mercury
thermometer during each exposure.
F. Records Retention
All raw data and th; final report will be stored in the archives of
Haskell Laboratory for Toxicology and Industrial Medicine, Newark,
Delaware, or in the DuPont Hall of Records, E. I. DuPont de Nemours and Co., Inc., Wilmington, Delaware.
RESULTS
A. Exposure Conditions and Associated Mortality
tofl|Bp Tiber temperature ranged between 18-34C dunnojexposures to
^jand between 27-35C during exposures
Wide temperature
ranges are not expected to affect.mortality in nose-only exposures.
Each test atmosphere contained a distribution of particles of
various sizes, including both small (smaller than 1 urn) and large
I
(larger than 10 urn) particles. The average geometric standard deviation
for each particle size distribution was approximately 2. Table I shows
j
total atmospheric concentration, particli- size distribution and
associated rat mortality for each exposure. Data are grouped from
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Page 4 of 14
exposures with similar particle size distributions. The increasing mass median aerodynamic diameters indicate a shift in the particle size
distribution from smaller to larger particles.
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HLR 110-85
^S6 5 of 14
Table I
At;mospheric
1ot al
Part 1C ulate Cone en tration
mg/m3)
djBHIE^JH Charac:terization and Assijciated IRat L>iposures
% b^' Weight. of Partic1<'s with Aercdynamic; Diameter 1(iss than 1, 13 U1Ti 5.2 urn 3.1 urn 1.1 urn
Mortali ty from
MMID" Deaths
A. ^^^^^^1
^16 + 6.6
99
92
73
21
1.7 urn 0/6
s42 7 ' 7.8
99
95
79
30
1.6 urn 3/6
7 25
98
93
77
21
1.9 urn 6/6
330 + 11G
95
75
44
8.1
3.4 urn 6/6
58 + 42
76
45
18
3.0
6.6 ijm 0.'6
77 7 5.3
86
52
20
4.9
5.8 urn 0/6
170 7 40
80
51
22
2.3
6.0 urn 4/6
i 1 B.
>
9.4 + 1.9
24 7 21
66 7 48
110 7 72
110 7 43
140 7 34
48 + 10
72 7 8.7
110 7 56
.190 7 88 '^320 7 56
390 7 100 520 7 140
57 + 18
84 -*- 31 190 7 33
<360 7 33
610 '7 82 520 "7 42
87
76
68
98
93
81
98
93
77
99
96
83
92
-6
53
98
89
62
67
50
34
78
52
32
73
50
29
88
60
26
89
56
27
77
48
22
79
54
28
57
29
10
70
43
23
63
39
19
74
46
22
71
41
18
75
42
17
56
1.1 urn 0/6
37
1.7 urn 6/6
26
2.1 urn 6/6
34
1.7 urn 6/6
20
2.9 urn 6/6
13
2.7 urn 6/6
18
5.2 urn 0/6
\7
5.1 urn 0/6
13
5.5 urn 0/6
5.8
4.7 urn 0/6
3.9
4.9 urn 5/6
1.6
5.4 urn 4/6
2.9
5.5 urn 4/6
5.0
9.7 urn 0/6
7.4
6.6 urn 0/6
4.8
6.9 urn 0/6
2.5
5.6 urn 3/6
1.1
7.1 urn 6/6
1.3
6.1 urn 6/6
a
Mass median aerodynamic diameter. Nebulizer and cyclone were heated during generation.
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HLR 110-85 Page 6 of 14
?
B. Estimated Lung Deposition and Associated Animal Mortality
The fractional deposition of particles within the respiratory tract depends in part on the particles' aerodynamic sizes. However, literature sources vary widely in their estimates of-the size-limits of particles able to be inhaled and to be deposited into various regions of the respiratory tract. Further, data indicate that deposition varies widely amoung individuals and amoung species.
' The Environmental Protection Agency has adopted the following
criteria to define the approximate size-limits of particles which may deposit into the various regions of the human respiratory tract: particles smaller than 15 urn can be Inspired and deposited throughout the respiratory tract; and particles smaller than 2.5-3.5 urn (nose and
mouth breathing, respectively) are expected to deposit predominantly in the alveolar region. Deposition of particle?; smaller than 3 urn is similar in rats and humans. Deposition data in rats for particles larger than 3 urn are not available.
To investigate the relationship between--HHoxi city and particle size, the following assumptions have been made:particles smaller than 3.1 urn will provide predominantly alveolar deposition, particles smaller than 13 urn'(including particles <3,1 urn) represent total respirable particulate, and particles larger than 13 urn will not be inhaled. The 3.1 urn and 13 urn size-limits were chosen because they are the experimental cut-points provided by the cascade impactor used in these tests which most closely approach the EPA criteria.
For each exposure, the atmospheric concentrations of particles smaller than 3.1 and 13 urn were estimated by multiplying the total atmospheric concentration by the mass pe-'cent of particules smaller than
these cut-points. As shown in Table I, within groups of similar
particle size atmospheres, mortality generally increased with increasing concentration. Further, as particle size distributions shifted towards larger particles, the concentration needed to cause death increased. The purpose of back-calculating the atmospheric concentration of particles smaller than T' and 3.1 urn was to investigate whether the apparent decrease in tr.--': j can be explained by the inability of a large fraction of these .it ospheres to either be inhaled or be deposited in the alveolar region.
ForHlUPaxposures, ^ne atmospheric concentration of particles
smaller t'nan 3.1 urn was most closely associated with animal mortality; regardless of total atmospheric concentration and MMD, as the
concentration o^articles smaller than 3.1 urn increased, mortality increased. ForfBHxposures, as the concentration of particles
smaller than 3.rumTncreased from 32 to 58 mg/m , mortality increased. However, one exposure containing only 19 mg/m of particles smaller than 3.1 urn caused 6/6 deaths. The deaths at this concentration were
"IBHIIBUf ,unexDected^nd^he cause of death is difficult to explain. For both total respirable paniculate (all particles smaller lar^^^im^dT^not have a clear dose-response relationship with
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HLR 110-85 pggg 7 ^ ^4
mortality; as the concentration of total respirable paniculate increased, a corresponding increase in mortality was not observed.
Table II presents the atmospheric concentration of particles
smaller than 13 and 3.1 urn and associated rat mortality for representa tive exposures. Data for all exposures are presented in Appendix I*
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HLR 110-85 Page 8 of 14
&4
Table II
Atmospheric Concentrations of Particles Smaller Than 13 and 3.1 urn and Associated Rat Mortality"
Atmospheric ,
Concentration (mg/m )
Mortality
Data Calculated From
Particles smaller than 13 urn;
65
0/6
42
3/6
140
4/6
72
6/6
Particles smaller than 3.1 um;
15
0/6
33
3/6
37
4/6
56
6/6
77 mg/m- @ 5.8 um MMD 42 mg/m" @ 1.6 um MMD 170 mg/m' @ 6.0 um MMD 73 mg/m @ 1.9 um MMD 77 mg/m, @ 5.8 um MMD 42 mg/m, @ 1.6 um MMD 170 mg/m, @6.0 um MMD 73 mg/m @ 1.9 um MMD
Particles smaller than 13 um:
80
0/6
120
0/6
170
0/6
24
6/6
65
6/6
100
6/6
Particles smaller than 3.1 um;
32
0/6
36
0/6
49
0/6
19
6/6
51
6/6
58
6/6
110 mg/m3@ 5.5 um MD
190 mg/m, @ 6.9 um MMD
190 mg/m^@ 4.7 um MMd
24 mg/m, @ 1.7 um MMD 66 mg/m' @ 2.1 um MMD 110 mg/m @ 2.9 um MMD
110 mg/m3@ 5.5 um MMD
190 mg/m- @ 6.9 um MMD 190 mg/m- @ 4.7 um MMD 24 mg/m, (? 1.7 um MMD 66 mg/m, @ 2.1 um MMD 110 mg/m 0 2.9 um MMD
Atmospheric concentrations were estimated by multiplying the total
atmospheric concentration by the percent by weight of particles smaller than 3.1 and i3 um, respectively.
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HLR 110-85 Page 9 of 14
C. Clinical Observations
In general, very^few clinical j^gns were observed in rats that survived exposure toQ^--------------JL
During or immediately following both lethal and non-lethal exposures, some rats In several groups had test material on their faces and heads and had a diminished startle response. Most rats exposed to lethal concentrations had labored breathing, and a few rats exposed to
lethal concentrations had red nasal and ocular discharges, ruffled fur, decreased activity and pallor. A few rats exposed to non-lethal
concentrations had red nasal and ocular discharges.
^WI------B^ ^ During thejecovery period, most rats which survived exposure to slight weight loss (less than 5%) for 1 day after exposure, and had no major clinical signs. However, a few rats had greater than 51 body weight loss, facial discharges, diarrhea, wet perineum, ruffled or discolored fur, hair loss and labored breathing.
For^Bflexposures, most deaths occurred during exposure or 1 day post exposure,although a few rats died between 2 and 8 days post exposure. For|1------nexposures, most deaths occurred from 1 to 2 days post exposure, with the latest death occurring 6 days post exposure.
Rats that died lost approximately 7-151 of Initial body weight 1 day
after exposure, and continued to lose weight until they died. Clinical signs for rats that died included labored breathing, facial discharges, limpness, ruffled or discolored fur, wet or stained perineum, diarrhea, pallor and lethargy.
DISCUSSION
Based on total atmospherlcMncentcatlon, the Approximate Lethal
Concentrations for both^j|B------------Qincreasewdith increasing particle
size:
MMD
ALC
1.6 urn 6.0 urn
3
42 mg/m, 170 mg/m
1.7 urn 5.6 urn
24 mg/m3
360 mg/m
Although pureg^Qppeared to be more toxic thanQ----^in the smaller
particle size range, both materials were considered extremely toxic when administered as highly respirable aerosols. When the particle size distribution was shifted toward larger particles, these materials were considered moderately to highly toxic.
^^s^---------""681
HLR 110-85 Page 10 of 14
The apparent decrease in toxicity with larger particle sizes is best explained by considering the fraction of the test atmosphere expected to
deposit in the alveolar region. Regardless of total atmospheric
concentration, the concentration of particles smanAc^Jjan 3.1 urn was most
closely associated with mortality. Except for one f^xposure, as the
concentration of particles smaller than 3.1 urn increased, mortality
increased. explained.
The cause of death in the out-lying j|fxposure cannot be --=' y
CONCLUSION
^^J^ly^d^fih^^ehcfoicnonddiilti'ons of this test, the Approximate Lethal Concentrations
of----U^^B^Jincreased as particle size distribtuions shifted from
smiaaNrteerrttooTalarrggeerrpapra1rticles. Regardless of total atmospheric concentration, the atmospheric concentration of particles expected ro enter the alveolar region was most closely associated with mortality. However, one exposure to ^Hpaused death at a much lower concentration than was expected, and the ?auseof death in this exposure can not be explained by this model.
Calculation described in Sierra Instruments, Inc., Bulletin 7-79-219IM, Instruction Manual: Series 210 Ambit-nt Cascade Impactors and Cyclone
Preseparators.
?
Air Quality Criteria for Particulate Matter and Sulfur Oxides, External Review Draft No. 2, Office of Research and Development, U. S.
Environmental Protection Agency, February, 1981.
Acknowledgement:
in this study.
Bruce A. Burgess and Rudolph Valentine also participated
Work by:
ffafe^ T IAAA^rt^^
Robert T. Tuner
Technician
dax^ fi. ^U^^-^ S^Ck^- c"
Steven C. Carpenter^^A^
Tepcrhnniicnian
T
Study Director;
r ^ 3 5 g<3u
A f^ .
Laura A. Kinney Chemist
Approved by:
^a^. QJ^^^ ^I'l^
Nancy" C. Chromey.wi.b.'
Section Supervisor, Acute Investigations Section
LAK:sg1:1.2
Date Issued: April 1, 1985 Study Initiated/Completed:
Notebooks;
9/27/83-2/7/84
Haskell Laooratory Report No. 110-85 Number of pages in this report: 14
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HLR 110-85 Page 12 of 14
Appendix I
Atmospheric Concentrations of Particles Smaller Than 3.1 and 13 inn and Associated Rat Mortality
1. Concentration of particles smaller than 13 urn
Atmospheric
.,
Concentration (mg/m )
16 44 66
42 140
72 310
Mortality
0/6 0/6 0/6
3/6 4/6
6/6 6/6
Data Calculated From:
16 mg/m3@ 1.7 urn MMD 58 mg/m- @ 6.6 um MMD
77 mg/m" @ 5.8 urn MMD
42 mg/m30 1.6 urn MMD
170 mg/m'- @ 6.0 urn MMD 73 mg/m3@ 1.9 urn MMD
330 nig/nr @ 3.4 urn MMD
2. Concentration of particles smaller than 3.1 urn
Atmospheric
.,
Concentration (mg/m )
10 12 15
33 37
56 150
Mortality
0/6 0/6 0/6
3/6 4/6
6/6 6/6
Data Calculated From;
58 mg/m3@ 6.6 urn MMD
16 mg/rn. @ 1.7 urn MMD 77 mg/m 0 5.8 urn MMD
42 mg/m30 1.6 urn MMD 170 mg/m- 0 6.0 urn MMD 73 mg/m3@ 1.9 urn MMD 330 mg/m- @ 3.4 urn MMD
1. Condentration of particle'' smaller than 13 urn
Atmospheric
o
Concentration (mg/m )
8.2
32 32 56 59 80 120 170
Mortality
0/6 0/6 0/6 0/6 0/6 0/6 0/6 0/6
Data Calculated From
9.4 mg/m3@ 1.1 urn MMD 48 mg/m- @ 5.2 urn MMD
57 mg/m, @ 9.7 urn MMD 72 mg/m- @ 5.1 urn MMD 84 mg/m- @ 6.6 urn MMD 110 mg/m- @ 5.5 urn MMD 190 mg/m- @ 6.9 urn MMD 190 mg/m- @ 4.7 urn MMD
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HLR 110-85 Page 13 of 14
Appendix I (cont'd)
Atmospheric Concentrations of Particles Smaller Than 3.1 and 13 inn and Associated Rat Mortality
B. |jj(cont'd)
1. Concentration of particles smaller than 13 urn (cont'd)
24 65 100 110 140 270 280 300 390 410 430 460 820
6/6
24 nig/in3@ 1.7 uni.MMD
6/6
66 lug/in, @ 2.1 um';MMO
6/6
110 mg/m- @ 2.9 unTMMD
6/6
110 ing/m- @ 1.7 um;WMD
6/6
140 nig/in:- @ 2.7 uni;'MMD
3/6
360 nig/in:' @ 5.6 uriilMMD
5/6
320 ing/in:, @ 4.9 um^Mb
4/6
390 ing/in, @ 5.4 uni';MMD
6/6
400 ing/in" @ 2.5 unrMMb
4/6
520 mg/m- 0 5.5 urnMMD
6/6
610 ing/in- @ 7.1 urn MMD
6/6
620 nig/m- @ 6.1 um'tlMD
6/6
900 nig/m @ 2.6 inn MMD
3, Concentration of Particles Smaller than 3.1 urn
Atmospheric 3 Concentration (mg/m )
Mortality
Data Calculated From
5.7 6.4
16 19 23 32 36
49
19 51 58 79 86 86 87 91 110 110 150 260 570
0/6 0/6 0/6 0/6 0/6 0/6 0/6 0/6
6/6 6/6 6/6 3/6 5/6 4/6 6/6 6/6 6/6 6/6 4/6 6/6 6/6
57 mg/m- @ 9.7 urn; MMD 9.4 mg/m- @ 1.1 urn, MMD 48 mg/m- @ 5.2 urn MMD 84 mg/m, @ 6.6 urn'MMD
72 mg/m- 0 5.1 unipMMD
110 mg/m- (? 5.5 unrMMD 190 mg/m, @ 6.9 unrMMD" 190 mg/m" @ 4.7 urn: MMD
24 66 110 360 320 390 140 110 610 620 520 400 900
mg/m30 1.7
mg/m, @ 2.1 mg/m- 0 2.9 mg/m- @ 5.6 mg/m, @ 4.4 mg/m- I? 5.4 mg/m- O 2.7 mg/m- @ 1.7 mg/m" @ 7.1
mg/m- @ 6.1 mg/m- @ 5.5 mg/m- @ 2.5 mg/m (? 2.6
urn MMD urn MMD urn MMD urn.MMD urn MMD urn MMD urn MMD urn;MMD urnJMMD urn MMD urn MMD urn MMD urn MMD
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?^e"S'^ 14
