Document Rp7wQ4xzv9641BJKqeXVRZY7k
FINAL'REPORT ACUTE INHALATION TOXICITY STUDY IN RATS
T-2991!T
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Submitted to 3M COMPANY February 2, 1981
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LABORATORIES
ANAERICA, INC.
SPONSOR:
3m COMPANY
MATERIAL:
T-29911T
DATE:
February 2, 1981
SUBJECT:
FINAL REPORT Acute Inhalation Toxicity Study in Rats Project No. 154-157
1. SUMMARY The test material, T-22911T, was evaluated for acute inhalation toxicity In rats. Exposure to the test material for four hours at a nominal concentration of 6.57 mg/liter caused sniffing and preening. After 60 minutes of pressure, all animals appeared normal and remained so until the end of the study. Most exposed animals lost weight by post-exposure Day 1, but subsequently recovered. All animals gained weight by the end of the study. Gross pathology findings at necropsy revealed a higher incidence of lung lesions In exposed males, but there were no consistent findings indicative of a compound-related effect. Microscopic examination of fixed lungs, livers, kidneys and all grossly abnormal tissues also failed to reveal compound-related histomorphologic changes.
li. OBJECTIVE The purpose of this study was to assess the acute inhalation toxicity in rats to a single 4-hour exposure to T-29911T in conformance with proposed FIFRA guidelines (40 CFR Section 163-81-3, August 22, 1978). The study was initiated November 25, 1980, and terminated December 10, 1980.
154-157
'"HAZL=C:N
LABC)RATORIES AN*PCA@ INC
Ill. MATERIALS AND METHODS
A. Test Material The test material,T-29911T,a yellow liquid, was received November 19, 1980,
and stored at room temperature. -Information on the methods of synthesis, stability, as well as data on composition or other characteristics which define the test material are on file with.the sponsor. B. Animals and Animal Care*
A total of 20 healthy appearing rats, equally divided by sex, of the Sprague-Dawley descended strain, obtained from Charles River Breeding Labora-
tories, Inc., Wilmington, Massachusetts, were selected from a stock pool of 24.
Animals not selected for study were returned to the stock pool. The animals ranged in weight from 228 to 335 grams when selected, and had been quarantined
for at least 19 days prior to selection. Throughout the study, the animals were
individually housed in stainless steel wire-mesh cages with food (PurinaS Lab Chow #5001(o)and water (via automatic lick valves) available ad libitum except during exposure. A 12/12-hour light/dark cycle (6:00 A.M. to 6:00 P.M.) was maintained by automatic timer throughout the study. The exposure was conducted during the light phase. Rats were used in this study because they have historically been used in safety evaluation studies and are required by FIFRA regulations. C. Groups
The selected 10 animals per sex were assigned to two equal-sized groups by use of a table of random permutations of 16 (Cochran, W.G., and Cox, G.M., Experimental Designs, 1957) as follows:
These animals were maintained in animal care facilities fully accredited by the American Association for Accreditation of Laboratory Animal Care.
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LABC)RATORIES
ANAERICA. ff4r-
Group No.
Number/Sex
Treatment Target Nominal Concentration
1
5
Air Control
2
.5
5.0 - 5.5 mg/L
D. Exposure Conditions Group 2 was exposed to T-29911T aerosol in a 100-liter glass and stainless
steel chamber operated dynamically at a constant airflow of 16.7 liters/minute. Group I was "exposed" In a chamber to air only. The compound was generated for 240 minutes as an aerosol by use of a Solo Sphere nebulizer at an aspi,rator setting of 100% and operated at an airflow of 3.5 liters/minute.
The aerosol was introduced into the turret top of the chamber with make-up air at 13.2 liters/minute. The exposure was 4.0 hours in duration (Plus 30 minutes to allow for chamber purging). E. Exposu_re_Monitoring 1. Aerosol Concentration
a. Nominal: The nominal concentration of the exposure was calculated by dividing the total weight of T-29911T aerosolized by the total volume of air passing through the chamber during aerosol generation.
b. Gravimetric: During exposure to T-29911T, samples were collected on Gelman Metricel OM-450 filters via a probe extended to the approximate breathing zone of the rats. A sampling rate of 10 liters/minute was used. Four 15-minute samples were drawn, starting at 60, 115, 170, and 225 minutes after exposure initiation. Weight gain on each filter divided by total volume of air sampled (150 liters) was used to calculate each gravimetric concentration.
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AKAERCA.
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CORRECTED
c. Analytical.:The above Gelman DM-450 filters were shipped in sealed
glass vials, packed on dry lee In air-tight packages, to the sponsor
for determination of the weights of active material collected.
2. Particle Size Distribution: Prior to and twice during exposure,_s4mp_les-were----collected with Andersen 4-stage Mini-Sampler cascade Impactors In order to
determine the aerodynamic particle size (mass) distribution of the nonvolatile component of the aerosol in the chamber at the rat's breathing zone. Samples
were obtained starting at 60 and 170 minutes after exposure Initiation for 15-minute durations. A sampling rate of 1.4 liters/minute was used. The mass median aerodynamic diameter (MMAD) and geometric standard deviation
(ag) of the mass distribution was obtained graphically after converting the dry weight gains on each stage and backup filter to cumulative percentages
of total weight collected and plotting these againstthe stagecutoffs (in
microns) on log-normal (probit) paper. Values at the 50th and 84/50th (or 50/16th) percentile were taken as the mass median aerodynamic diameter and geometric standard deviation, respectively.
3. Temperature and Relative Humidity (R.H.): Temperature and R.H. In the con-
trol chamber were measured with an Abbeon Model M2A4B temperature and R.H. gauge and recorded every 30 minutes. Temperature and R.H. in the chamber varied from 69*F to 75*F and 45% to 53%, respectively. F. Biological Observations
1. Toxic Signs: Continuously during exposure and twice daily for 14 days post-
exposure, all animals were observed for signs of toxicity, abnormal behavior, or unusual appearance. Noteworthy changes were recorded.
2. Body Weight: All animals were weighed and body weights recordedprior to exposure and on Days 1, 2, 3, 4, 7, and 14 post-exposure and prior to
necropsy.
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LAEMORATORES
ANERCA, PJC-
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CORUCTtrO
G. Necropsy
All animals were killed on Day 15 post-exposure by exsanquination under pentobarbital sodium anesthesia. Complete necropsies were performed and gross pathology recorded. Special attention was paid to the lungs and upper respiratory tract. Lungs, liver, kidneys, and all.grossly abnormal tissues were fixed in 10% neutral buffered formalin.@ H. Histopathology
Preserved tissues were embedded In Paraplastle@ sectioned at 5-6 microns, slide mounted, stained with hematoxylin and aosin, and examined microscopically by a board-certified veterinary pathologist for indications of compound-induced changes. 1. Data Storage
A]) raw data, specimens, and the final report will be stored In the archives of Hazleton Laboratories America upon acceptance of this final report.
A. Aerosol Concentration
111. RESULTS
1. Nominal Concentration: A total of 26-33 grams of T-29911T aerosol were
dispersed into the inhalation chamber with a total air volume of 4008 liters.
The nominal concentration was, thus, calculated to be 6.57 mg/liter of air
of T-29911T.
2. Gravimetric Concentration: Results of the dry weight gravimetric concentra-
tion determinations are shown below In Table 1.
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LABORATORIES
ANAERICA.
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154-157
CORRECTM-
Table 1 - Gravimetric
Chamber
Concentrations
of T-2991[T Aerosol(dry weight).
Sample Initiation
Tm-i-n.T-
60
115
170
225
Weight Collected
(mg)
8.89
9.13
10-74 10.86
Sample Airflow -7L-Y-
150
150
150
150
Gravimetric Concentration
(mg/L)-
.059 o61
.072
.072
3. Particle Size Distribution: Results of the particle size determinations
are shown in Table 2. The majority of the mass collected was in the res-
pirable size range (i.e., less than 5.0 microns).
Table 2 Particle Size Distribution Data.
Stage No.
1 2
3 4
F
MMAD ag
Cutoff Size p
>4.7 <4.7
<3.3
<2.1
<0.65
Preliminary Weight Collected mg
18 .11
.25
.26
.03 .83 2.6
2.00
B. Mortality No animals died during this study.
Sample
WerghtCollected
mg
No. 2
Weight Collected
mg
.03
.05
.05
.07
.35
.52
.15
.32
.00
.02
.58 2.4
.98 2.35
1.33
1.47
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LABORATORES
ANAERC.A. t-Jr-'
C. Toxic Signs 1. Group 1 (Air Control).: All animals in this group exhibited normal appearance and behavior throughout the study.
2. Group 2 (6.5 mg/L): After one minute of exposure, all- cin-iincIit -were --observed--
to be sniffing the air, and after five minutes, all animals were preening. All animals appeared calm and normal after 30 minutes, and all animals appeared normal after 60 minutes and throughout the remainder of the exposure. All animals exhibited normal appearance and behavior throughout the 14-day post-exposure observation period. D. Body Weights Individual body weights with group means (S.D.) are presented in Table 3. Most Group 2 animals lost weight after exposure, but subsequently recovered. All animals gained weight by the and of the study. E. Gross Pathology Gross pathology findings at necropsy are presented in Table 4. There was a higher incidence of lesions In the lungs of Group 2 males, but there were no consistent findings indicativeof a compound-related effect. F. Histopathology Microscopic evaluation revealed lesions of chronic respiratory disease consisting of peribronchial and perivascular lymphold hyperplasia and focal pneumonitis characterized by focally thickened alveolar walls and a pleocellular inflammatory infiltrate, including alveolar macrop@ages. These lesions were essentially comparable in incidence and severity between control and treated rats with the exception of the control females, in which lesions were slightly less prominent. Histopathologic incidences are presented in Table 5.
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LABORARORIES ANEFICA WW--
Microgranulomas and minimal nonsuppurative pericholangitis were noted in liver sections. Kidney sections revealed minimal to slight focal interstitial nephritis. Hydrometra was present in uterine sections from three females. Lymphoreticular cell hyperplasia was present in the various lymph nodes examined and was accompanied by congestion and pigment-laden macrophages -in the mediastinal lymph node of one control female. Foci of agonal hemorrhage were noted in thymic sections of three rats.
In conclusion, microscopic evaluation of hematoxylin and eosin stained sections of lung, liver, kidney and unusual lesions from control and treated rats from an acute inhalation study with T-19911T failed to reveal compoundrelated tissue alteration. Spontaneous disease lesions were essentially comparable in incidence and severity between control and treated rats.
Submitted
by:
z
WILLIAM B. COATE, Ph.D.
Director
Inhalation Toxicology Department
8
154-157
Animal No./Sex
47931cf 47932d 47933<t 4?'934cf 47935(?
Mean @-S.D.
'7Q362 47337? 47138-7 479391 L-7940@
Mean S.D.
Table 3 Body Weights (grams).
Pre
321 341 -'-0 )22 323
325 8.8
Day- I
319 341 325 324 323
326 8.5
Day 2 Day 3
GROUP I
324
331
343
348
323
328 -
328
334
329
338
329 8.0
336 7.8
Day 4 Day 7
324 348 331 335 340
336 9.i
344 369 '1244 5-7.2 357
353 10.4
271 2.3i 141.56 2m03 2 7S
264 18.6
271 238 256 290 280
267 20.5
276 240 2r.6 291 275
268 19.8
'73 240 :Z60 291 283
269 20.1
268 236 257 278 279
264 17.8
2:8o 236 260 291 275
268 21.3
Day 14
382 408 381
86 395
390 11.3
286 243 266 286 284
273 18.8
47941(@ 47942e 47043d 47-9044d' 47945(?
Mea.-i S.D.
479462 479472 479482 479492 479502
Mean s.D.
336 350 345 350 340
344 6.2
248 239 242 249 278
251 15.5
319 336 319 345 328
329 11.2
237 233 241 256 265
246 13.6
GROUP 2
329 347 333 350 337
339 9.0
337 353 342 358 346
347 8.4
237 239 242 258 278
251 17.3
241 239 242 256 282
252 18.1
338 354 343 357 342
341 8.2
243 238 241 254 270
249 13.1
355 376 362 383 363
368 11.4
246 239 244 260 279
254 16.2
382 414 401 423 405
405 15.4
254 242 259 270 289
263 17.8
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Table 4 Gross Pathology Findings
1 OBSERVATIONS
GROUPS
2 d,
NO GROSS LESIONS
2/5 2/5
1/5 1/5
LYMPH NODES
Tracheal bronchial:
appear enlarged
Cervical:
left appears enlarged
Mediastinal: left appears reddened left appears enlarged and granular left appears enlarged right side appears enlarged
THYMUS.
purple foci right side reddened left side reddened
115 1/5 1/5
115 1/5
1/5
2/5
1/5
1/5
1/5
LUNGS
All Lobes:
numrous pale areas
1/5
scattered white foci on surfaces
scattered red areas
patchy appearance
Right Diaphragmatic Lobe:
dorsal surface has two black foci consolidated linear area that did not perfuse
Right Apical Lobe:
did not perfuse
Left Lobe:
1/5 1/5
1/5
1/5 1/5
1/5
anterior tip has dark reddened area surrounded
by a pale area
1/5
Intermediate and Left Lateral Lobes:
scattered red areas on ventral surfaces
1/5
CECUM Red area, probable injection site
1/5 1/5
KIDNEYS
Dark eort ico - medullary area Dark zone in cortico - medullary area
1/5
1/5
1/5
UTERINE HORNS
Appear distended with clear fluid Appear thickened
- 10
1/5
1/5
1/5
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No.
147931 147932
47933 m 47934 47935
4793'161
47937t7o 3
479388 !L
47940
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477941
147942
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147945
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rvlimal
No.
47799331 477993322
7933 477993334 47934
479935
147-.9773d963367 47937 -n
14793811 1479391 147940
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1477941
147799944422 0;
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4794551 0 lrn 794
1 4779944661 7r4749747 47948 47948 47949 In
47950
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PERSONNEL
154-157
STUDY DIRECTOR: WILLIAM B..COATE, Ph.D.
b
REPORT PREPARATION: 4REWRY 4(. -FAB IAN-111, B.S.
PATHOLOGIST..% DEBORAH BANAS, O.V.M., M.S. Diplomate,American College of
VeterinaryPathologists
INHALATION SUPERVISOR:
Z442:2
ROBERT J- KAftY
TECHNICIANS: ALLMAN, D. TRAMMEL, HENRY, GRIFFEY, REDDEN, HAND, KIMBLE, K. TRAMMEL, BOWERS, UDY
13 -
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LASMATCRlr=S
ANAERCA. W'44C
OFFICE OF QUALITY ASSURANCE
ProjectTitle: Acute InhalationToxicity Study in Rats Project No.: 154-157
quality Assurance inspectionsof the study and review of the final report of the above referenced project were conducted according to the standard operating procedures of the Office of Quality Assurance and according to the general requirementsof the Good Laboratory Practice regulations that were issued on December 22, 1978, by the Food and Drug Administration for compliance on and after June 20, 1979. Findings from the inspectionsand final report review were reported to management and to the study d-irectoron the following dates:
Inspections/Review
Findings Reported
Inspector/Reviewer
Study - 12/10/80 Final Report - 1/26/81
12/11/80 2/2/81
E. Prins K. Hogan
A-
e
G @r r
@M@annaggree@r
Office of Quality Assurance