TRL STUDY 0032-006
                            Conducted for:
                      RESEARCH TRIABLE  INSTITUTE
                           P.O. Bex 12194
             Research Triangle Park* North-Carol1na 27709
                                  by:
                  TOXICITY  RESEARCH  LABORATORIES. LTD.
                         510 West Hacklcy Avenue
                      Nuskegon.  Michigan   49444
                  Rat Oral  Subchronle Toxlclty Study
                               Compound:

                            Normal  Butanol
Start of Test (pretreatoent):                   August 19.. 1985

Interim Necropsy:                              Oct^r'7;and,8.;1985,

Final Necropsy:                                NB*^r;25;iiid; 26.:i985

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           TOXICITY  RESEnRCH LOBORttTORIES
 Document Title    |



 ROT ORflL SUBCHRONIC  TOXJCITY  STUDY  (DRRFT)
    Chemical* Catogory.
 N-BUTONDL

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T8L Study 1032^006
The  purpose  of  the study was *e evaluate the toxic Ity of normalfcutanol ,.
In • rat subchronlc toxlclty study.
     f
Four  groups  of Mlc  and finale rats  (30/sex/group) were  dosed orally
once  dally with 6.  30.  125. or 500 ag/kg/day of compound for 6 weeks
Miit 11 the  day of the Interim necropsy.  After the Interim sacrifice,  §11
surviving  rats were dosed dally until the final sacrifice.   Body weights
and  food  consumption were recorded weekly and the rats  were observed at
least twice  dally  for aortal 1ty and evert .signs of toxlclty.  Ophthalmo-
loglc  examinations were  done  during the  pretreataent  period and  again
during  week 13.  Blood  end urine  for ellhlcopathologlc valuation were
collected  from  a fifth group  of  10 rats/sex  prior  to  initiation of
dosing, from all  surviving rats  scheduled for the,1nter1m sacrifice,  and.
from the first 10 rats/sex/group at the final sacrifice.   The first iq
•ale and 10 female  rats  from  each group were scheduled for necropsy on
day 43 or 44 and  all  remaining rats oh day 92 or; 93.  -ififoss -pVstiijortem
examinations were done on all  rats.   Organ weights- were taken^fron  rats
 sacrificed  on  day  92  or 93.   Aftar  the}. filial  sacriflcei. a; cdmpjete
 Mstopatholcglc wamlnatlon  was doiie on all rats in the Control:, .land
 high-dose groups, en livers.  Mdney.s, e*d hearts from: the low- and  nld^
 dose groups. .and  en all gross lesions.  In addition,  a 'Mstopathoroglc
 examination was  done en any. rat  fpund/toad:  er:tacr1f1cad.r1n'ce«tce)nls;;
 The .only  une i|i1 vojcaj , ef f ectt; Produced 7Dy.;fH)nnaKbuttnol :wer^ ^ttx1aland;

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TRL Study §032-006               - «  •
             at the  100  ag/kg/day dose level.  The aaxlBum weekly Inci-
dence  was  32X end 291,  respectively, but ataxla OF hyj»ac1t1v1ty were
not  Been as trttatoent-related  signs  until  the final six  weeks of the
study.   Wo  treataent- related  clinical  signs  were seen at the 30 or the
125 Eg/ kg/day dose level.

Sody  ealght «nd  food  consumption values were similar  for control and
all  treated groups.   Elo treataent- related effect was  observed at the
ophthalnoscopk  examinations  or  In  gross or alcroscoplc evaluation of
the  tissues.

Three rats were found dead or  sacrificed In extremis,  but  these deaths
were wot due to  administration  of the  noraal bwtanol .

 At the  Interim  clinical  pathologic  evaluation,  red blood  cell  count
 (RBC),  packed cell  volume (PCV). and hsneglobln (MSB)  averages of the
 §00  f»B/ka/day  *»«  group  feaales   ware  SS below control  averages.
 Although these  differences  were statistically   significant,  they were
 small and  no  differences between the parameters were  observed 1n the
 sales ef  the  Interla evaluation or between control and treated groups
 ®f either sex at the final evaluation.  Therefore, even  1f  the lower red
 blood cell parameters 1n the 500 eg/kg/day fesaales were  an  actual treat-
 »sm> related affect, 1t was stall end transitory.

                                CONCLUSION

 Oral  ©dBlnlstretlen ef woraal batanol at  500 ag/kg/day poduced ataxla

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TRL Study 0032-006              - 111 -                    DRAFT

and  Hypoectlvlty  at  a ofixloun weekly  Incidence rate  of 32  and  29X,
respectively.  A  sUflhtly (5J) 1:-er (conparad  to  controls)  red blood
sell eeynt (RBC), packed cell »olune (PCV). and heaoglobln (IGB) concen-
tration  j^e&ent 1n  the 500 eg/kB/day *se trouP Resale* at the Interim
evaluation anly isay have teen treataent^felated.

eio  treatwent-related  aifect  nas  ooserved at the 30 BB/*fl/
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TRL Study #032-006

                               Table  of Contents
I   Introduction [[[   *


U   Methods

     A  Test Material [[[   J
     B  Animals and Husbandry «.««••«•••••••««••«*•«•••*•«••*«'•«•••••••••••   Z
     C  Compound Administration ....••.•••.••.....•••••••.••••.•••••••••'r.«   3
     D  Clinical Observations ..............................................   J
     E  Qphthal wol ogy ............................. • ....................... •   *
     F  Clinical Pathology .................................................   *
     6  fecropsy ,.«••••.••••••••••••••»••••••»•••••••••••••••••••••••••••••   •
     H  Histology [[[   f
     I  Statistics [[[   f
     J  Data Retention ........................ . ............................   »


HI  Results

     A  Test Material [[[   f
     B  Clinical Signs ...... .. ............... • .............................    »
     C  Body Weight* Weight Change and Food Consumption ....................   10
     0  Ophthalmology ...... ••••••••••••••••••••• ..... •••••••••••• ........ ••   10
     E  Clinical Pathology .................................................  }*
     F  Observations at Necropsy .........•...•••••••••*••«••••• ........ ••••  12
     6  Organ Heights [[[  \*
     H  Mlstopathology [[[  l3
 IV   D1«eyss1on [[[

 V    Conclusion .. ................... • ........... ».tt« ................. •••••  15

 VI   References [[[  l6

 VII  Tables

     Table  1  Incidence of Clinical Signs ....... • .........................  17
     Table  2  firoup Mean Body Height, Height Change end
                    Food Consumption «e«.«.»<>eoBo.«.«..«o.e»««»««««»«»«««»»«'  20
     Table  3  Msnatolegy Suaaary •••••••• .............................. ••••  ||
     Table  4  Serua Chonlitry Susanary eo» ..... eo.e«*. ..... e« ....... • .......  91
     Table  5  Hffliatology Values ............. •••••§«• ..... «••!••••••• ......  37

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TRL Study 0032-006
DRAFT
     Appendix A  Compound Assay Results
     Appendix B  Ophthalnologlc Evaluation
     Appendix C  Clinical Pathology Methodology
     Appendix D  Individual Rat Reports (Cross and H1stop»tholeg1c)
     Appendix E  H1stoaccountab1l1ty Report
     Appendix F  Protocol find Amendments.
     Appendix G  QAU Statement

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    TRL Study 0032-006           • I •
DRAFT
I    Introduction
     The  purpose of  this  study «*  *> evaluate the toxlclty of  noraal
     feutanol  1n a «t  subchronlc toxldty study.  In  order  to  assess the
     tox1colo01c  potential  sifter  both 6 weeks and 13  weeks  of  tfoslng, an
     mttrta sacrifice at  the 6 week  Interval  MS  Included.8  The  oral
     route of administration was used  because this Is the  probable route of
     human exposure.

     This study was  conducted 1n accordance with  the protocol  (Appendix F),
     the  Standard Operating  Procedures  of Toxklty Research Laboratories
     (TRL) and 1n compliance with 6ood Laboratory Practice Regulations for
     •oncllnleal Laboratory Studies.1*   Procedures pertinent to this  study
     ere described herein.

 II  Methods
 A.  Test Material
     Normal   butanol   (lot*  3597   KVVE) was   purchased  from  toerlcan
     Scientific  Products,  Romulus, Michigan.   Samples of each  dose concen-
      tration were saved  during weeks 1. 4, 6. 10.  wd  13 «nd  taken  to the
     ttoskegon County Hastewater Treatment Facility for chemical analysis at
      the laboratory ywter  the direction of Or. Avi Joshl, Physical  Chemist.
      Duplicate staples were  Mrt to Mr. John Maney. ERCO, a division of
      Enseco, Inc.. IBS Alwlf* Brook Parkway, Cambridge,'Massachusetts  for
      referee analysis.
           Protocol Change 04. effective »/10/B5.
           Federal Register, Vol. «3, ft. M7. Pitt II, tecember 22, 1978.
           pp. S99B6-60020.

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     TRL Study W32-006            - 2 -
DRAFT
B.   Animals  and Husbandry
     The rat  was chosen as a test system because of Us established useful-
     ness  1n  toxlcologlc  studies  end  as  a  pharmacologle  asodel.   One
     hundred-fifty three Bale end one hundred-forty seven female rats* (45-
     55 grains) aged 22-23 days Arrived en August 12. 1985 and were housed
     Individually  1n  wire-bottom  cagesb.  Filtered oinlclpal  water0  and
     Purina Certified Rodent Laboratory Chow** were available ad libitum.
     This feed has been tested by the ranufacturer for contaminants, none
     of which were present at levels that would be expected  to affect the
     outcome  of the  study.   An  declination  period  of  7 days  prior to the
     pretreafenent  week was  si lowed.  During  the  acclimation period,  the
     rats were observed  with respect to general health  and  any rat with
     evidence  of   disease  or  physical  abnormality   was  discarded.    A
     clean/dirt^  corridor system was In effect.  Room  air was filtered and
     humidity  (average 47.6X.  * 9.2)  and  temperature (average 70.2°F.  t
     2.2)  controlled.  The  temperature value  was calculated  using  the
     dally high and  low value. *1  standard deviation.  One  value  per day
     was used  to calculate the  humidity  average.  A 12 hour  Ughf.lZ  hour
     dark cycle was controlled automatically.
          Charles,, River Breeding  Laboratories.  Inc.. Portage.  Michigan,
          Crl: CDR(SD)BR.
          Rats were housed  In  accordance with recommendations contained In
          BHEH Publication  No. 78-23 (WIH): Revised  W78.  B6u1de  for the
          Care  and Use of Laboratory  AnlMls."   During the  acclimation
          period the rats ©ere boused 3/eage.
          Mater  used  at TRL  analyzed  periodically  for the  presence  of
          contaminants  as  defined by the  Snvlrwaental  Protection Agency
          •national  Interim  Prleary  Drinking  Mater   Regulations11  Code
          af Federal Regulation. Title  4C~Protect1on  of  Environment  Part
          l4i.ll (B) end 141.11.  Records  retained at TRL.
          Let  0s   §tay02-85-3E,  Aug01-B5-2E.   Aug07-85-lB.  AuglO-B5-2D.
          Aug2l-|5-$D. Purina Lab Chow-12T, Checkerboard Stpare. St.Louls.
          Gn\   £31Bo.

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    TRL  Study  #032-006            - 3 -
     The first day  of the  pretreatment  week was August 19.  1985.  At  the
     Initiation of the pretreetoent week  the  rats were assigned randomly to
                              /
     groups  (30  rets/m/group  end  a  fifth  group  of 10/sex)  using  a
     computer  printout.8   The rats  were  Individually  Identified by  toe
     clipping.

£.   Compound Administration
     Administration  of the  test  ooterlal  began  on August 26.  1985.   The
     rats scheduled  for the  Interim  sacrifice were dosed dally for 42 or 43
     days and the rats scheduled  for the final sacrifice were dosed dally
     for 91 or 92 days.
          Normal  Butanol                      Animal Numbers
     Croup     (e«/kq/day)           Male                     Female
     —_e                  interim         Final       Interim       final
     I            0        001-010         OTP530     031-040      04TB60
     II           30        061-070         071-090      091-100      101-120
     III         125        121-130         131-150      151-160      161-180
     IV          500        181-190         191-210      211-220      221-240
     V         Baseline     241-250                     251-260
     The amounts  administered were  based on Individual weekly body weight
     values.   Fresh solutions of the compound  (In  delonlzed water) were
     prepared weekly  and dosed orally at a volume of  10 ml/kg.  Controls
     received delonlzed water at  the sane volume.  A plastic syringe and an
     18  gauge  ball»t1pped seta!   dosing  cannula ensheathed 1n  a number 8
     French catheter were used to administer the solution.
     8    Randomization printout programed  by Dr. John Quiring. Associate
          Professor  of Natheaatlcs  end  Computer  Science. Department  of
          Hathsnatlcs  and Computer Science, Brand Valley  State Colleges,
          Allendale. ttlchlgan.

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     TRL Study 0032-006            - 4 -
                                                               DRAFT
D.   Clinical Observations
     Body mights and food consumptions  were  recorded weekly and the rats
     were observed  at least twice dally  for BortaWy and clinical effects.

E.   Ophthalmology
     All rats received an ophthainoseoplc  examination during the pretreat-
     rent   period   and  week   13  by  a  veterinary   ophthalmologist*.
     Ophthalmolealc examinations were conducted  on  all  rats  1n  a darkened
     room «1th  an  indirect  ophthalmoscope.   Seven rats  that had an  eye
     lesion during the pretreataent period were not Included In the study.

F.   Clinical  Pathology
1.   Sampling
     Blood and urine samples were  collected from the 10 sale and 10 female
     rats 1n group V prior  to  Initiation of dosing. Blood was obtained at
     the time of necropsy from  all  surviving rats scheduled for the Interim
     sacrifice and from  the first  ten  rats/sex/group at the final sacri-
     fice.  Urine  MS  collected  In eetabollsm cages 3-5 days prior to the
     scheduled  sacrifices.   The  rats  were  anesthetized  with  C02,  the
     thoracic cavity was opened and blood was  collected by cardiac punc-
     ture.  A necropsy was then done on  each of these rats (except those
     1n group V) end the tissues preserved.
      *    Exams  perforeed by  M.  F. teller. O.V.M.. M.S.  end
           Diplomats,  taeHcan  College  »f Veterinary  Ophthalmologists,
           Hlchlgan State University. East Lansing. Michigan.

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     TRL  Study 0032-006           - 5 -
                                                              DRAFT
2.   Tests Par-formed
     The following tests* were performed:
                         .
             Hemoglobin (HCH). Been cell honoglobln concentration
             (MCHC). total  and differential  leucocyte counts (WBC),
             estlnated platelet count (PIT).
ChBHstry;    eUallne  phosphatase (AU Jjos), Mood I urea nitrogen
             (BUN),   plutonate    Rrruvate   transaminase   I»PT;.
              '                                    aOT.   lucose
                             ,
                        I'utiMta  oxelecetate  JTMHMIMM  iaOT).  glucose
                        [Glue),  total  protein  (TP). albumin  (Mb). »/6 ratio
                        (calculated),  fllobulln  (calculated). toU  bll rubln
                        (Tot.  bill.),  wdlua  (Ma),  potassium  (K), chlorldg
                        (Cl).  calcium  (Ca).11   inorganic  phosphate (FJoi)i
                        carbon dioxide (TC02),  total «enw cholesterol  (Choi).
                        creatlnlne.
                              »pedf1c  gravity,  glucose,  protein,
                                     urobHlnogen.  »1croscopy  of   *ed1ment.
 6.    Hecropsy
      K necropsy  was  perfortneii  on  all  wrvlvlng rats of the first 10 males
      and 10 females  from  each  dose  group on day 43 or 44  of the study.  On
      days 92 or 93  the remaining rats were necropsled.   Rats found dead
      *ere also necropsled.   Interim and final sacrifice rats were anesthe-
      tized with 002 and  exsanguinated  by  cardiac  puncture.  The cranial.
      thoracic  and  pzrltoneal cavities  were  opened and the  contents examined
      aweroscoplcally.   The  organs  and tissues listed 1n  section H were
      reaoved from  each eMul and  preserved.   Lungs  were Inflated with
      foraaHn via the  trachea.    t>es with attached optic nerve from  all
      *    See Appendix C for clinical pathology aethodology.
      *    Protocol  Change  II.  tffectlve  8/15/85; Protocol  Change  «,
           effective B/13/85.
      e    Protocol Change «4, affective 9/10/85.

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    Tfc. Study 0032-006            - 6 -                        _.  ^  m
                                                              DRAFT
     rats  Ml led at the  Interim and final  sacrifices  were  preserved  In  a
     oodlfled  Zenker'K fixative.  The testes with  attached ep1d1dym1des
     from  all rale  pats were  preserved 1n  Bouln's fixative.   All  other
     tissues  were preserved 1n  10<  neutral-buffered foraalln.   Feet were
     preserved  vlth the  tissues for  positive Identification of  the rat.

     Prior to fixation at the final  sacrifice only,8 the following organs
     ttere  weighed:   brain,  heart,  liver,  spleen,  kidneys,  testes  with
     ep1d1dym1des. and ovaries.   After fixation,  the adrenals and thyroids
     01 th  parathyroids were weighed.*  For paired organs, the organ weight
     was the  combined  weight of right and left cambers  of the pair.  Organ/
     body  weight ratios were determined for each tissue. No organ weights
     were  taken  on rats found dead.

H.   Histology
     The full tissue microscopic examination listed below was done  on the
     control  and high-dose rats, en one rat sacrificed 1n extremis,  and  on
     those found dead.  Also, livers, hearts, and kidneys  of low- end w1d-
     dose   rats, and  all  gross Isslons  seen at  necropsy were  examined
     Blcroscoplcally.

     As  tissues were trlensd.  the  presence  or absence  of  tissues  and
     lesions «as noted.  The tissues  were  placed in Tissue Tek®6 cassettes
     that  Mere  labeled  with study  number,  rat  number and the cassette
          Protocol Change #4. effective 9/10/85.
          Lab Tek Division, Miles Laboratories,  Inc., NipinrlUe,  XL 60MO.

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    TRL Study W32-006
                   -  7 -
    number.  Th«y  were then processed  on  a  Fisher Scientific H1stomat1c®a

    or  an AO TP/BOod?6   After  processing,  the tissues were  embedded  1n

    paraffin using a Tissue T«k*«snbedd1ng system.  They were sectioned at

    6-6 alcrons, aounted on nuabertd slides and italned with hewtoxylln

    and eosln.


    The following tissues were placed on sequentially numbered slides as

    follows:
Slide Number

     I
     I
     2
     2
     2
     3
      S
      s
      6
      7

      1
      8
      9
               Tissue

heart and attached aorta (a longitudinal section)

lung* (Actions from the caudal and left lobes)
trachea (a cross-section)
 t                          nonglandular .sofhage.l are.
through tXe area of the cardiac  glands  Into the area of the
f Sndll fllands end another wctlon from  the duodenum throuoh
the  pylorlc sphincter Into the area of  the pylorlc glands)
salivary glands (sections of the subllngual and »and1bular

8MllS1ntest1nes (a separate cross-section of  the duodenum.
jejunum and 1l0un)
colon  (a cross-section)
liver  (sections from the left and right lobes)
pancreas (a separate section 1n addition to sections
cwunonly. attached to the viscera)
spleen (a  cross-section)

                           «•
 adrenal)
   ewl   attached «pt1e ftsrve)
 thyroids snd |«i-athyro1ds             e
 tlSreelc spinal  eerd (a erws-sectlra)
 liaabar spinal cord (a cross-section)
           fisher  Scientific  Products.  SM01  Industrial  Road.  Livonia. MI
           68150.

           fesrlcen  Optical  Scientific  XnstnB*nts Division,  lutfalo. W
           14215.

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TRL Study #032-006
                     -•-                     DRAFT
                    Tissue
g          tt -M. «M*MM- of
ii .      E    i:         arAs ss
13
H
B. «tc.  SMMM ... .11 othr «r»» '«"<>«
 At  th.  Mitologle  w»1».t1.n.        «««  ««  «r«""'  «*«"
 sioderate, « • 8»v«r«, ami S • txtrwt.
  H. tody «1.ft. *- «««n,pt1.». .H.lcop.th.lo.fc. .m. «!••
  *u ««  t.»t-  for  »»...«ity *  »i— » "rtlrtt>'
  (St«!  «* T.«-1.. UN).  M th. 4.U «« fpund to be ho.09.Mous.
             Nt—
  .ttMttlcrt ,1,n1f1e.»c. by th. MM tf MMt (MMtt. 1«»).  M

  th. *tt  ««  1M< «ot to *  I-MM".  «• "tl>011  " S1"
          DuBiwtt't) aa npleywl (6111.
   Beta Retention
   HI  *t. 1«1».1«  .M1M,
MM.
           .t  t«»»c1ty  IMM     .. It...
                . Weh1,.n «M4 for .t 1.«t . y..«.  Nfore
   8   Protocol enrage A. effective W16/85.

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HI. Results
A.   Test Material
     The  results  of the  analysis of  saaples  analysed  by the Wastewater
     Treafenent Facility and  by SRCO ere given *« Appendix A.   Stability and
     econcentration were found to to acceptable.

8.   Clinical Signs
     The Incidence of clinical effects  Is given In Table 1.

     Treetmsnt-related Jtajc1a-f4rst-««p»aped -4«-tbeJdgh=dose .group dur1.n$
     <*sek S_.  At&x1a and hypoactlvlty occurred  1nf recently  during  weeks 9
     Qnd jo.  These signs Increased to a weekly Incidence of 32 and 29S for
     ataxla  end hypoactlvlty.  respectively, at  ueek 11 and  continued  at
     epproxlmately  the same freqiency during weeks  12 and  13.   Onset  of
     ataxla  and hypoeetlvlty was about 2-3 alnutes after dosing and dura-
     tlon was less  than one hour.
     Other clinical signs observed did not appear to be directly related  to
     treatment.   Three rats  died  during the study.   Two  of these  deaths
     were the result of the  rubber catheter slipping  off  the wtal  dosing
     eannula.  During week 4. a catheter lodged 1n the esophagus of a high-
     dose female  and the  rat died  of apparent  asphyxiation before  the
     catheter could te raaovcd.  During roek 7. a high-dose oale swallowed
     © catheter.  It was sacrlf 1c«d  too days later often It teeam apparent
     that the  situation was tdvarsely affecting  the haalth  §f  the rat.
     This accident and the occurrence  of  ^activity, salivation, labored
     respiration, and/or retching over  a  S day  period 1n  another high-dose

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    TW. Study 0032-006           -  10 -
    rat account  for-  the clinical  effect*  observed  1n  the high-dose  group
    during »««**  6 «i»d *•   * aid-dose sale died during week 1.  It  began
    exhibiting hypo»et1v1ty. mac tot ton, and labored respiration on day 4
    and uas  found dead en day 6.  H1stopatholog1c  evaluation of this rat
        i be  dUcussed later 1n the report.
    Dark  yrlne  and  rales occurred  1n a   high-dose sale  during  week 8
    following  a eage related accident.  This rat was normal  In appearance
    »1th1n  three  days.   During  week  7.  a  2  en  diameter  tissue mss
    appeared  1n the left axillary area of  one low-dose female and  a 3 on
    diameter  oass appeared 1n the right axillary area of  another low-dose
    female.   WeUher aass  Increased  In tlze  and both  disappeared  before
    study termination.

C.  Body Height. Height Change  and Food Consumption
    The body weight, weight  change and food consumption  values  are given
     1n Table 2.
     No  treatment- related effect was  present on body weight, weight
     or  food consumption.   Statistically  significant differences between
     control  and treated  group seen  might change  and food consumption
     would  eeeur sporadically, but no trend was @b served  and  total body
     weight averages for control and treated groups  were stellar  throughout
     the study.

     Ophthalmology
     The results ef the ophthalraloglc evaluations are given In Appendix B.

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    TRL Study 0032-006            - XI
     An ophthalmoscoplc examination  uas  performed prior to  1n1t8t1on  of
     dosing  and all animals with eeular abnormalities  were  Identified and
     discarded.   Another ophthalmoscoplc  examination  teas performed during
     »»ek  i3.  The  pathology  observed was considered  to fee within normal
     Halts  for the age, sex, and strain of th« animal.

E.   Clinical Pathology
     Results  of  the el1n1cop«tho1eg1c evaluation  are given  In Tables  3
     through 7.

     Only one alteration In clinical  pathologic parameters occurred that was
     suggestive of  a  treatment- related effect.  At the  Interim  sacrifice.
     SBC  (p«S 0.05).  PCV  (p* 0.01), «nd  KGB (p« 0.01) averages In  the
     high-dose females  were IX less  than  the corresponding  control  aver-
     ages.  The RBC and PCV (p <0.05) averages for the middle-dose females
     were also slightly  (4X and  31.  respectively)  below  those of the con-
     trols.   However,  RBC.  PCV,  and  K5B  averages  were similar for control
     and  treated groups  of Bales  st the  Interim evaluation And for control
     and  treated groups of both sexes at  the final evaluation.
     Other  §mist1cally  slgnUNeant  differences  between  control  and
     treated  f?rw»P  averages  wcurved fcut  they were satl!. occurred In one
     eex  and  at ene evaluation enly, and there was w> dose response rela-
     tionship.   ?tesB ttey  «ere  aot eensldepad  to fee  treeteant^related.
     They eere:  ft  lower  (p 
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    TRL Study 0032-006            - 12 -                          DRAFT

    higher  (p< 0.05)  relative  segmented wsutrophll  count and  a lower
    (p €0.05)  relative lymphocyte count  1n  the low-dose females at the
    final  evaluation, and higher  (p«0.05)  urine pH values 1n the low-
    dose  Bales at  the  Interim end low-dose  females  et the final evalu-
    ations.

F.  Observations  at Hecropsy
    Observations  made at necropsy  ere given 1n Table 8.

    No treateent-relattf  lesion  was  observed  In  gross necropsy at  the
     Interim §r final  sacrifices  or ef the rats  found dead or sacrificed
     1n extras.   The  lesions  present were  those commonly  observed  1n
     laboratory rats  end they were present 1n  control and treated groups
     at similar  f regency  or war* one  tlae occurrences.  The enlarged
     uterine horns are related to  the  stage of the estrus cycle.

     Three pats died  during  the  study.   Wo gross lesions were seen 1n rat
     d 224.  aat $ 202 (which had  a catheter 1ri  Its stomach) had  dark areas
     ®n the glandular oucosa of  the stomach.   In  rat  0 133 (alddle-dose),
     the  left lobe  of  the lung was  red  and  the cranial and nlddle lobes
     Here shriveled.
           Heights
     Relative  and  absolute group Bean organ might values are  given  1n
     Table 9.

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    TRL Study 0032-006           - " -                       foR A PT
                                                              •^ ix f^\ i B

    There was no apparent treatment-related effect on organ might values.
    The  only  statistically  significant  difference  between  control  and
    treated  group  averages  was a  slightly  (p« 0.05) higher thyroid
    *e1ght average  1n the hlgh-dose sales.  Ho dose response  relationship
    «ros  present, as  toe  ebfolute  thyroid  weight*  were stellar for all
    three  treated groups  ef  Bales.  Moreover,  they  were only W  above
    the  control average.  Thyroid  weight averages of the treated femiles
    were  not above those of the controls.  Thus, this difference appears
    to be  e chance  occurrence rather  than  a treatment-related  effect.

N.  Histo pathology
    The results  of  the Mstopathologlc examination ere given  In Table
     10  and  Appendix  0.  The MstoaccountablHty  report  U given  In
     Appendix E.

     No  treotsnent-related  lesion  was   observed  at  the hlstopathologlc
     evaluation.  Tne  lesions that were  observed were one time occurrences
     or were present  In the  control end treated groups  at A similar fre-
      «8iency.   The diffuse subacute lyaphadenltls ef  the aandlbular lymph
     node  was vUable  grossly  as red or enlarged lymph nodes.  This 1s a
     eoswonly  ebserved lesion  In laboratory  rats.   The cause ef death of
      the  Bid-dose ret («33) that died during the study was determined to
      fee a savaging accident since e perforated esophagus was found at the
      hlstopathologlc  exenlnatlon.
 IV   Discussion
      At tba littrfai evaluation. f®C, PCV. and KB averages 1n the hlgh-dose

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TRL Study «32-006            - 1« •                          DRAFT

females were 58  below control  averages.  The differences were statis-
tically  significant  but the biological significance 1s Questionable.
All  three parameters arc  closely  related,  so a  decrease  1n the RBC
count  »oul(!  be   eRpected  to  result 1n a  decrease  1« «V  and MSB
values  also.  Moreover,  the observed differences were wall  and no
difference  between these  parwwters  was  *««n In Bales at the  Interim
evaluation  or between control and treated groups of either sex at the
fine! evaluation.  Thus, even  1f the decrease 1n RBC count  (and hence.
PCV and  MSB)  was a  true  treatnent-related  effect, 1t was  swell and
 transitory.

 One Bid-dose and too high-dose rats  died during the study,  but none of
 these deaths  was due to  administration  of  noraal butanol.   The two
 high-dose  rats  died because  the rubber  catheter slipped  off of the
 dosing cannula during dosing.   In one case  (ret « 224). the  catheter
 lodged In the throat and esophagus,  thus cutting  off  air flow  Into the
 lungs.  The rat died of apparent asphyxiation before  the cannula could
 t*  removed.   Ko gross or Mstologlc lesion relating to the  
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    TRL Study #032-006          •«-                     DRAFT

    result tf esophagi towge at dosing.

V   Conclusion
                    «  .  ».«»                    •'« "*
               .   A
                             Mil »1«  (PCV). Md
                           tki MO vyttf
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    YRL Study 0032-006           - W -
VI   References
     Dunnett, C.M. (WM). filsetrlcs 20.482-491




     •111. J.u. (W77). f
           .
     Wew York.

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Data produced
            (Month)     (Day)     (Year)          Camera Operator
Place   Syracuse       New York
     (City)            (State)
                                                ASVSTEK
                                                  corp

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