United States                EPA-600/7-82-038b
               Environmental Protection                '
               Agency                   May 1982	
v°/EPA        Research and
               Development
               ENVIRONMENTAL ASSESSMENT OF
               A LOW-EMESDN OIL-FIRED
               RESIDENTIAL HOT WATER
               CONDENSING HEATING SYSTEM
               Volume IL Data Supplement
                Prepared for
               Office of Air Quality Planning and Standards
                Prepared by
                Industrial Environmental Research
                Laboratory
                Research Triangle Park NC 27711

-------
                 RESEARCH REPORTING SERIES


Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination  of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:

    1. Environmental Health Effects Research

    2. Environmental Protection Technology

    3. Ecological Research

    4. Environmental Monitoring

    5. Socioeconomic Environmental Studies

    6. Scientific and Technical Assessment Reports (STAR)

    7. Interagency Energy-Environment Research and Development

    8. "Special" Reports

    9. Miscellaneous Reports

This report has been assigned to the INTERAGENCY ENERGY-ENVIRONMENT
RESEARCH AND DEVELOPMENT series. Reports in this series result from the
effort funded under the 17-agency Federal Energy/Environment Research and
Development Program. These studies relate to EPA's mission to protect the public
health and welfare from adverse effects of pollutants associated with energy sys-
tems. The goal of the Program is to assure the rapid development of domestic
energy supplies in an environmentally-compatible manner by providing the nec-
essary environmental data and control technology. Investigations include analy-
ses of the transport of energy-related pollutants and their health and ecological
effects;  assessments of, and development of, control technologies for energy
systems; and integrated assessments of a wide range of energy-related environ-
mental issues.
                       EPA REVIEW NOTICE
This report has been reviewed by the participating Federal Agencies, and approved
for  publication. Approval does not signify that  the contents necessarily reflect
the views and policies of the Government, nor does mention of trade names or
commercial products constitute endorsement or recommendation for use.

This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.

-------
                ACUREX TECHNICAL REPORT TR-81-78/EE
ENVIRONMENTAL ASSESSMENT OF A LOW EMISSION OIL-FIRED RESIDENTIAL
               HOT WATER CONDENSING HEATING SYSTEM
                  VOLUME II:  DATA SUPPLEMENT


                           April 1982



                       Acurex Project 7600

                       Contract 68-02-3188



                               For

                 EPA Project Officer - R. E. Hall
                    Combustion Research Branch
              Energy Assessment and Control Division
           Industrial Environmenta Research Laboratory
          Research Triangle Park, North Carolina  27711
                               By
                          C. Castaldini
                        Acurex Corporation
                 Energy & Environmental Division
                         485 Clyde Avenue
                 Mountain View, California 94042

-------
                                 CONTENTS

Section                                                                 Page

   1       Introduction 	     1
                References	

   2       Preliminary Equipment Calibration  	     2

   3       Complete Furnace Operating Data  	     7

   4       Sampling Data Sheets 	    39

   5       Analytical Laboratory Results	    56
                5.1  Fuel Analysis Report   	
                5.2  Particulate Gravimetric Analyses and
                     Mass Emission Calculations 	    59
                5.3  Sulfur Analysis Report and Emission
                     Calculation	    64
                5.4  Trace Element Analysis Report  	    66
                5.5  Organic Analysis Report  	    88
                5.6  Bioassay Analyses Report 	   112

   6       SAM IA Work Sheets	   180
                                    iii

-------
                                SECTION 1
                              INTRODUCTION
       The purpose of this Data Supplement is to document data in greater
detail than was practical in the final report (reference 1-1).  It is
intended to provide sufficient detail for researchers to perform their own
analysis of the data.  Readers are referred to the final report for
objectives, description of the emission results, interpretations, and
conclusions.
       The data supplement contains the following information:
       •   Preliminary equipment calibration — calibration data for the
           dry gas meters used to sample flue gas for both EPA Method 5
           and Source Assessment Sampling System (SASS)  trains
       t   Complete furnace operating data — furnace water and flue gas
           temperatures throughout each burner-on/burner-off cycle;
           cooling water flowrate, and thermostat settings
       0   Sampling data sheets — emission data obtained with continuous
           monitoring instrumentation:  operating data tables for EPA
           Method 5 (for particulate mass emissions), EPA Method 8 (for
           S02 and S03 sampling), and SASS (for particulate, trace
           element, and organic sampling)
       a   Analytical  laboratory results -- ultimate analysis of
           distillate fuel  oil  used in the test program:  laboratory

-------
           analysis reports on participate levels by gravimetric
           analysis; sulfur by turbimetric analysis; trace elements
           by Spark Source Mass Spectronetry (SSMS) analysis and
           Atomic Absorption (AA) analysis; anions by ion chromatography;
           Total Chromatographable Organic (TCO) emissions by onsite Gas
           Chromatography (GC); speciation of organic emissions by Gas
           Chromatography/Mass Spectrometry (GC/MS); biological assays of
           the extract and water discharge samples
       t   SAM IA work sheets  — Source Assessment Model  IA evaluation of
           emission and effluent data  to determine the  potential
           hazard posed by both flue gas and  water discharge samples

                        REFERENCE  FOR  SECTION 1
1-1.    Castaldini,  C.,  "Environmental  Assessment of  a Low-NO
       Residential  Hot  Water Condensing  Heating System Burnifig  Distillate
       Oil, Volume  1:   Technical Results."

-------
            SECTION 2
PRELIMINARY EQUIPMENT CALIBRATION

-------
         --7-8 C
Dm
BeroKtrlc rVetiure 3o-l0
                                                                                                            £•0
            2.0
            2.2
            2.4
            2.6
                                                                     HI
                                                                   JT

                                                                             3J-
                                                                                     S&-0
                                                                                              73
                                                                                                       TSr
                                                                                    _2t.
                                                                                                       73
                                                                                  '.•*?
                                                                                                            n-o
            3.0
            3.2
            3.4
            3.6
                                          33Uu£2h.
                                                       e>*B>m
                                                                             "ga"
                                                                   JLfc.
                                                                                               73
                                                                                             73
                                                                                              JZ2.
                                                                                                       75
                                                                                                                       3
-------
Oit«
toroMtrtc leisure

Je*1ent Twpcraturt
                       Orifice Meter

                       Orifice Nignehellc

                       rrtairy Calibration Utter

                       Control nodule*
                                                                                         Operiton

                                                                                         Met lulb Teaperature
                                                        HETER CALIBRATION  DATA
 Orifice
 Manometer
 (1n.*9.)
Orifice
Magnehellc
Prlwry
 Meter
Dry Test
 Meter
  Cts Volim
PrlHry Meter
  fiei Voluw
Dry Teit Meter
                                                              (ft.1)
                                                                                              Teaperatvre
                                                                               Prlwry Meter
                                                               Inlet.
                                                              TP1  ™
                                                           Outlet.
                                                           TP» «'F>
                                                                                               Dry left Meter
                                                                                                    Inlet.
                                                                                                             Outlet,
                                                                                                                               Tine
                                                                                                                                t
                                                                                                                         J
             0.2
             0.4
             0.6
             1.0
                                               JS- 1
                                                                                                                      70-0
                                                                                                                              3/-0
             1.2
             1.4
             1.6
             2.0
                                                                          •70
                                                  • 899
                                                                                   70
                                                                          ra
                                                                                           7/0
             2.2
             2.4
             2.6
                                             630-
                                                                          13.
                                                  ' Qll
                                                                                                     -TO
                                                                                                              -TO
                                                                                                                     7C-O
                                                                                                                                7/7
             3.0
             3.2
             3.4
             3.6
             4.0
             4.2
             4.4
            .4.6
            4.8
            S.O
                                                                                                              70
                                                                                                                      70.0
                                                                                                                           fit
                                                            IS-SS6
                                                                                                                                      Average

-------
Drt.

TIM

laroMtrlc Prttsun

Milcnt T«perature
                       OHflct Ntttr

                       Orifice Nagnenellc £o£3

                       Prtaery Calibration

                       Control Module  &}
                                                                 Operators

                                                                 Met ftilh Temperature
                                                       METER CALIBRATION DATA
 Orifice
 Manneter
    •

Orifice
fegrahcllc

   «„
Prlwry
 Meter

 4H
Dry Test
 Meter
                               (In. iq.)
  Gas Volim
PrUtary Meter
  Gas Volwe
Dry Test Meter
                                                  (ft.1)
                                                                                             Te^erature
                                                                              Prlury Meter
                                                  Inlet,
                                                    <*F>
                                                 Outlet,
                                                 V <
 Tlac
  t

(•In.)
*
            0.2
            0.4
            0.6
                                                                         *S-
                                                                                                             bl*
                                                                                                                              t-0
             1.0
             1.1
             1.4
             1.6
                                                         a> &•
                                                                Lt,
                                                                                                     •TO
                                                                                                  lit*
                                                                                                  US
             2.0
             2.2
             2.4
            2.6
                                                                                                    73
            3.0
            3.2
            3.4
            3.6
           tsp
                                                                                                                     7o
                                                                                                                   3.0
                                                                                                                                     I.OL&
            4.0
            4.2
            4.4
            4.6
            4.8
            5.0
                                                           /&-99S
                                                                                                                                      Average

-------
           SECTION 3
COMPLETE FURNACE OPERATING DATA

-------
                                                         FURNACE OPERATING DATA
Cycle
No.
±
Cooling Hater
Flowrate
(gpm)
X7





















V






















Thermostat
Sgtting
53





















V,






















Inlet Water
Tempgrature
A'fc
«P
*L
£<»
*(.
&{~
£(*
£(*
gfi,
SL
3L,
3t>

«*
XL

tL,
f(*
Outlet Water
Tempgrature
t,v
-II
-7k*
80

u-r

VI
Tank Hater
Tempgrature
?7
/o/

n^
//
-------
FURNACE OPERATING DATA
Cycle
No.
5.
Cooling Mater
Flowrate
(gpm)
/7




























tf
/7










Thermostat
Setting
t°C)
53














\















f3





	 s





Inlet Mater
Tempgrature
Sk
	 A-^ 	
•Via
f\l>
gt,
6<*
S(~
£(•
/,-£
£b
£<,
£<*
£<*>
srt,
S(»
g<»
A'L
ft.
*L>
*>'<-
Xi,
£<»
•?(»
Outlet Mater
Tempgrature
Sto
7?
79
77
la~
T+
Th
73
73
73.
71
7o
70
£,'1
iff
i-8
7/
77
»l
814.
»7

-------
FURNACE OPERATING DATA
Cycle
No.


Cooling Water
Flowrate
(gpm)
t-7





















\






















Thermostat
Setting
1C)
63,





















*





















Inlet Mater
Tempgrature
6V
S't,
46>
A'ff
*s
^r.-f
g*
4-;
f*s>~
fix
A'*
4V
5V
£n
6'*
5+
.$-*
56
SH-
*>•*
A'i>'
-<*.
.-fS
Outlet Water
Temperature
t*
97
<&
100
103
/Ov-
/o-o"
/oa
-
/>?P
«9
/JC>
«3
13-1
Hi
||^
//«.
//A
Exhaust Duct
Temperature
91
tf/
fiH
ea
g"a
frAt
sy*
7<1
t,*
W
c,a
t,i
L'i
6.7
LI
1*7
Ambient
Temperature
73.
73
LI
7JL
73.
73.
-71
«a
ffo
•76
7«
W
7^
•79
-73
7A
70
11
t,9
(A
M
if
. L>% '

-------
FURNACE OPERATING DATA
Cycle
No.
3






Cooling Water
Flowrate
(gpm)
/7






17


































V
Thermostat
Sgttinq
53





'
SIS

















V


















Inlet Water
Temperature
-rv
.£:>-
£->'
&?
5+
&*.

Z*
Sfy'
£»'
£.*
6t
£•*
£ X
5*
£Z
£*
&
£(*
&A
X.<

Outlet Water
Temperature
72
7/
7/
7o
73
7»
fi-3
(?*^
t> <9
^/
rV3
R
/o7
/o-f
/07
//3

yi,
/as
/9^
/37
!*.!
/*.*
/*i.y
/*!
f*U.
/o7
/fcCJ
/t,a
/6il
/a'7
/-T3
/+1
Exhaust Duct
Temperature
4-r
LI
66,

63
fc«
73
77
79
Sto
S*/

92

»_t
g,a-
S<~
e(~

<2*>
«l
78

Ambient
Temperature
L9
Li
6t>~
&•?

L*
t>L
^
1,7
AsT
t,3
t,t
^»
6,7 	
•y/ 	

6V
^y
7iT
8*3

«*^
. , •g'r- 	 ! 	

-------
                                                          FURNACE OPERATING DATA
Cycle
No.
H-


Cooling Mater
Flowrate
(9Pn)
/.7
















\i

















1.1



\




Thermostat
Setting
ro
tt.f
















\
















/
$3.tT



J



/
Inlet Hater
Tempgrature
S*
A'A'
SC
&'
5*f .
£if

**
£t>'
5»'
&»-
£*
A'i?
A~f>
AX
s.<
.<-','
^
--l'.*>"
Outlet Mater
Tempgrature
9/
%>
89
V*
Hi
XI
90
80
77
7-7
76,
"*¥•
~7*t-
7-3,
73.
7A
7/
70
7*
71
ff3
47
&}
Tank WatPr
Temperature
lu-L,
II+SL
/38
/3A~
f3S
/J?
/2C
/Stf.
,al
/if
//7
IIU-
//3
III
/0
-------
FURNACE OPERATING DATA
Cycle
No.

Cooling Hater
Flow-ate
(gpm)
/ 7





















*





















;
Thermostat
Setting
rcj
53.5"
















;







i







V







Inlet Hater
Tempgrature
&
f>
<$•*
4?
£<*
£<<>
S^
^

<£,
^
£i,
4?
Ji,
A,-

^
Outlet Hater
Temperature
9s
/

«
96
ffc.
«3
76,
3o

-------
FURNACE OPERATING DATA
Cycle
No.
£
Cooling Water
Flowrate
(gpm)
/.?







\







t
/.7












	 1













Thermostat
Setting
l°C)
53.iT







>







f
53.>

























f
Inlet Hater
Tempgrature
•f -4*
"
&'6
xv
A" 6.
.-ft.
S"
A'6
-)lo
J
5(~
-ft*
XL
A~(r
£?£.
£<*
XC, J
A'(>
-<^
.<6,
Outlet Mater
Temperature
77
7
*7y-
T3
7J
•ra
~7A
1i
7*
go
*±
V-J
£
n 3
Hi
/o9
IC,7
/CL.
ion.
/o3
lot
na
li 7
I3L3.
127
133.
/ft,
Al
73
^-r
75
7J
7<*
7^-
7^-
77
. 81

-------
FURNACE OPERATING DATA
Cycle
No.







Cooling Hater
Flowrate
(gpm)
1.7









































/
Thermostat
Setting
l°C)
6'3.5-





















V






















Inlet Water
Tempgrature
l,

Outlet Mater
Temperature
/on
99
t
t,t
7/
•73
7^
73
7/
71
71
6, a
Ambient
Tempgrature
ff7
9to
9/

-------
FURNACE OPERATING DATA
Cycle
No.
to







Cooling Water
Flowrate
<9P»)
/7





















•






















Thermostat
Setting
l°C)
$3.*





















\





















.
Inlet Water
Temperature
s L
ft*
£6.
*{„
ft,
A'C*
Si-
XL
*>'£,
*'<*
.•5V-
£-<•
ift.
ft.
£<„
£<„
-•*•£.
A"6>
A(~
-•4V."
-<(i,
£•&,
&
Outlet Water
Tempgrature
V3
79
SI
((*
33
ISf
JH-I
I+*
/*«
/o-y
AfV-
J.-S-I
S.-Z&
/.•fa
/v-w
/A.-T
M
A?«
A3*.
/.5?2
«^
Exhaust Duct
Temperature
£3
£?
ya
77
78
So
So
83
#a
£a
*^
Gu.
St+

-------
FURNACE OPERATING DATA
                                                                 10
Cycle
No.
7
Cooling Hater
Flowrate
(gpm)
X













7













/ 7














Thermostat
Setting
rci
53












\
.>












1
$3.*













/
Inlet Hater
Temperature
.5^
Zc,

£l*

<•/
<£,
<"£»
s" 7
•TC»
•5"6>
^"7
A' 7
.-i'i.
j£V
	 jffc. 	
Outlet Hater
Temperature
va.
ffn
So
79

77
7£


TA
73
7.3
7A
-j f
73
77
sa
&>6
»r
9£>
93
%>"
^
Tank Hater
Tempgrature
IS(o
J2U.
13 1
h&
117
//«- 	
//.-a
//o
/of
/a 7
/0(v
/O^4-
...tQA 	
,01
/O 1
/ok 	
n I ._
117
/J3.
13.7
J3I
I3i?
\ '+0
Exhaust Duct
Tempgrature
~9^
7o
•J5U
;*^-
•»*
•r3
TH
73
72
•ya
72
72
7«
73
^3
^,3
^
73
7-7
71
»0
SI
V2
S>3
Ambient
Temperature
°F
St»
80 	
f-3-
-79 	 .
ffS
S>0
S3
So
Sto
gy
Sb
s/
»a
7ff
•77
•7ff
7a
-7B
7A
-TU.
^r.<
•7A-
73

-------
                                                            FURNACE OPERATING DATA
                                                                                                                             /I
Cycle
No.







Cooling Mater
Flowrate
(gpm)
1.7





















\






















Thermostat
Setting
(°C)
53,5-












































Inlet Water
Temperature
if<~
XL,
.<7
£7
£(*>
£'&
.<&
.?>•"<"
Outlet Mater
Temperature
<9
iO7
J0*>
91
y<~

-------
FURNACE OPERATING DATA
Cycle
No.
9

Cooling Water
Flowrate
(gpm)
/7 .





1





/
//




























V
Thermostat
Setting
ro
53 if





\





f
5J.J?














V















Inlet Water
Tempgrature
A'f
A'fc
-4-6
A' 7
6'k
.:>'&
.«•&
.«»
-/
/M-7
Exhaust Duct
Temperature
72
74-
-73-
if?
7+
73
^
t>*
tH
73
77
•79
€f
So
«/
tei
8>a
?a
«3
»3
s?3
9/
tfo
Ambient
Tempgrature
ff»

f?3
79
8/
So
77
76
SO
79
7#
77
-78
79
78
79
77
7S
8*0

^/
«2
, tf& '

-------
                                                           FURNACE OPERATING DATA
Cycle
No.
4
Cooling Water
Flowrate
(gpm)
1.7









































i
1.7 •
Thermostat
Setting
(°C)
53. f








































y
vS"
Inlet Water
Tempgrature
£(.
	 i?(v
V7
^6
A- 7
x<~
Si
i3<0
< "7
i>(u
f?<0
*
IO +
Xo3
ioi
/oo
n
'03
Exhaust Duct
Temperature
7«
77
77
77
•w
76,
77
Vfc
7«
7B
75
7-^
73
7^
73
76~
7W-
73
7a
73
6,7
1,3.
tb
Ambient
Tempgrature
ft
t
gg
68
g-3
«^
96,
77
79

ffo
ga-
ro
o

-------
                                                         FURNACE  OPERATING DATA
Cycle
No.

Cooling Water
FloMrate
(9Pm)
1,7





















\






















Thermostat
Setting
t°C)
$5





















\






















Inlet Water
Tempgrature
S<~
A~ 7
g-C,
&<*
5<~
£r
.r<*
5" 6
« 7
.5*6.
5" 7
4'7
4' 7
<5"
A'C
Outlet Water
Temperature
go
«3
(?7
»^
ts.
1*.
3
/osT
to!
<97
<*<+•
V 1

-7V
Tank Hatfr
Temperature
/OS
//u.
/;9
/AM.
/^9
/.«
y.^7
IH-I
/•*-•*-
y^H
/X-^>
//r/
/v-S
/<*.i>"
/^-o
A? 7
/^^.
/J*/
/a9
/JG
/J3
A3/
//?
Exhaust Duct
Temperature
71
7t>
-r-r
-7<1
80
91
SI
81
«?a
»?,
S-^
83.
ffl
&o
-ft
73
~77
77
77
77
77
7b~
7&
Ambient
Temperature
fib
80
gsz
TV
«3
79
-r9
-r«
-79
»/
ff*^
87
93
«?o
y/
9»o
S>3
93
««•
9o
93

-------
                                                          FURNACE OPERATING DATA
PO
ro

-------
                                                         FURNACE OPERATING DATA
Cycle
No.

Cooling Water
Howrate
(gpn)
/,7





















•





















1
Thermostat
Setting
t°C)
56










































V
Inlet Water
Temperature
*><»
&(r
tffe
A"<~
i)*-
3~£

A"(*
5'6
4"<«
i'fc
£<»
f>-f
f*lr
\4<~-/)to
U '
\&
--TC
.<<;•
.Vt-
Outlet Water
Temperature
/o3
iO*4-
/0-*
loo
 _
' A&JxVfA/
7f
78
7-7
77
7<"
Tank Mater
Temperature
1*3
/•S3.
/.*•+
/S3
/«.
/.58
/3 !a"
/A2.
yao
7.27




//-3
II it-
US
Exhaust Duct
Tempgrature
€-3
!?^
ga
fto
77
ry
76?
Tff
78
77
77
77
77
-rt>
t*~
XT




73
7J
7^
Ambient
Temperature
ffo
7ff
8«-
83
ffa
»«»
(P3
S^
fia
ffa
82
ea
aa
»o
-77
•7f
) 	
/
/
/
77
7
-------
FURNACE OPERATING DATA
                                                              17
Cycle
No.
II







Cooling Water
Flowrate
(gpm)
/7





t





f
/,7














V














f
Thermostat
Sgtting
$*











I
6$














i














/
Inlet Hater
Temperature
ft,
5"<-
•^<^
6'<*
A~6
A-£.
.->'<«•
4'4-
6'^
€<'
s<.
,6'fc
5" 6.
.1-6.
5-6
A'd-
*"6
o'^-
A' 7
A'^-
A'6
A'4.
-S'^'
Outlet Mater
Temperature
7d>"
7*/
7.1i
73
7J
7/
70
7H
7?
*3
S-4
«S
7o
<*^
y.->"
'/«
/oo
A? 2
/o**
/o.^
/OAfc
«yy
<*>•
Tank Water
Temperature
ltd
icf)
IG-J
/OS
/04-
ios
/Ol
/at?
//O
11*4-
120
I2U-
/JS
i x:\
/37
/*/
y«.a
/«.^
/no
x^-3
/•af.-f
yn3
>M.<>
Exhaust Duct
Temperature
7^
73
•7^k
7a
ya
73
^3

-------
                                                          FURNACE OPERATING DATA
Cycle
No.

Cooling Hater
Flowrate
(gpm)
/ 7










































\J
Thermostat
Setting
t°C)
5
-ft,
•>'<>
£<*
A' 4
£~t,
J<~
S<~
rT6
i>'l>
.T4.
A-<&
A"^
A~«-
S'C
^"<^
A" 7
ri"~
9 +
ya
HI
&0
7V
7?
77
77
7.<
7^
7^
7V
73
7A
73
7/
71
71
Tank Hater
Temperature
/"**'
/v-/
/3«
/a-r
y.w
/39
/J7
/o
ea
f?7
9/
fite
s^
*&
•79
7tf
e^
&a
ff^-
s<*
»^a
9G
tf6.
»7
**
8--S"
S?/A
??7
S'^
. #3
ro
ui

-------
                                                           FURNACE OPERATING DATA
Cycle
No.
/£










Cooling Water
Flowrate
(91*)
/.7










































J
Thermostat
Setting
rcj
•Z5










































\)
Inlet Water
Tempgrature
£>' 7
&<~
*t,
<7
s 7
A' 7
A'6-
S-7
a-r
SI
,<-7
£7
*7
47
~f7
S7
*'(.
.6-7
A~&
X-7
£7
A' -7
.•T7
Outlet Water
Tempgrature
7t
<3o
«3
»7
*9
ft

»/
&o
79
?*f
77
Tank Hater
Tempgrature
iol
107
//a
J/7
/a.3.
/A7
/3i
/3H.
/3V
tn.3.
/4*o"
S+3
/•f-0
/37
t*3
S3&
/as?
I2H-
/aa
/\ao
us
///„
//*!.
Exhaust Duct
Tempgrature
6>7
71
7*4
77
78
79
79
fito
&o
8/
K2.
SI
79
St*
79
79
ffo
»/
7»
77
&>
Sfc>
S'o
Ambient
Tempgrature
80
?2
»a
9V-
g*a
0v
?&
ff6
i?7
SV-
*/
9a-

. «?/
ro
en

-------
                                                          FURNACE OPERATING DATA
Cycle
No.
/3
Cooling Mater
Flowrate
(gpm)
t,7













\













[
/.r






\






/
Thermostat
Setting
rcj
55-













V













1
fS






I






(
Inlet Mater
Tempgrature
6" 7
.<6>
f •*+
•5" 7
&'7
A'-T
.V7
«T7
ft"7
-T7
o-7
i>~ 7
*'t.
A' 7
X7
£7
T7
A'6
o'7
A' 7
A* 7
A* 7
*T 7
Outlet Mater
Temperature
7
7A"
7**
7-y.
•ra
/a
73
7f
7/
70
7o
Vt
i><*
i»<*
7f-
7%
«
1,7
-re,
73
7to
77
77
77
7^
Ambient
Tempgrature
^0
97
99
fiL.
Sfc
87
«5
8-r
ffi-
«a
»/*
H3
6>4
fi-3
83
?0
ff3
*2
.'SO
e-o
so
9^
. «/ '
ro

-------
                                                             FURNACE OPERATING DATA
Cycle
No.

Cooling Water
Flowrate
(gpnO
i,1





















\





















/
Thermostat
Setting
m
££~










































J
Inlet Mater
Tempgrature
A'C»
	 £-(c,
-5" 7
&
/o^
toy.
^7
9J
•?/
#9
*v
»A'
ft-^
«3
*/
80
TV
77
77
7t-
7-5"
7.T
-7.3
73
Tank Water
Tempgrature
/3k
Ht-O
/HM.
/«7
M?
/*-«
tt+U-
f*-c>
/*7
,3 +
/3i
/as
/3S'
/J*
yjy
y//
«/
«?0
7^
7-7
7V
7,f
7*
7S"
77
Ambient
Tempgrature
«/
«3
«3
»7
/oo
ion.
ma.
99
tOCf
/oo
9S
S-/
»8
i?9
9J
S7
S>£,
fft
84
»7
83"
fff-
. 94C '
rsa

-------
                                                         FURNACE OPERATING DATA
Cycle
No.
M-
Coollng Hater
Flowrate
(gpm)
1.7













/
t.7



























i
Thermostat
Setting
t°C)
SS






i






J
££


























v/
Inlet Hater
Temperature
A-&
4' 7
A" 7
S~(*
i>-t>
&<*>
*,'<*
6~<~
A' -7
*~t>
6' 7
5" 7
-5'&
*>' 7
6" 6
5-<^
6" 6
.5" 7
s-t
ft 7
4' 7
a- 7
A- 7
Outlet Hater
Tempgrature
•ra
7/
71
•70
•70
&ff
6t
&.K
-//
7^
go
fc-3
?
So
Sfc>
»o
6tf
t~~7
"70
73
76,
77
79
7
-------
FURNACE OPERATING DATA
Cycle
No.







»*
Cooling Hater
Flowrate
(gem)
'•7




















I





















//
Thermostat
Setting
l°0
*55*






































i
y
s*
Inlet Mater
Tempgrature
-6'7
.•>- 7
*<°
A'C,
A'<~
g~>
g (a
A' 7
^6,
g <~
£7
S7
s-r
A 7
*7
S-J
S7
t>'(*
S -7
s<*
f7
£7
4-6
Outlet Water
Temperature
?/
»8
<47
3£
S'S
S3
&/
So
-71
7S
77
~7l*
7:>
73-
73
73
72
73
7/
y/
70
iff
71
Tank Water
Temperature
/^/
/.^-7
yj^.
131
/as
ta.L,
/33
/a/
//s-
//4»
//y.
HA
/to
/o
-------
FURNACE OPERATING DATA
Cycle
No.

Cooling Hater
Flowrate
(9pm)
1,7





















\





















f
Thermostat
Settinq
t°C)
*Z$








































\
V
Inlet Water-
Temperature
•f L.

A" 7
•y 7
ft*
j-c.
t? (-
A~ 7
t>'7
A" 7
£•<-
*,-<*,
*:<-
A"<~
&7
i>~(a
A' 7
S7
$<*>
A-4
A- 7
S7
6't>
Outlet Mater
Temperature
Tfc
	 £o
8 <+
&7
e$
ft
V3
9 if
1 7
¥1
/oo
/CO
<*<.
93
9&
&8
&7
8*
«3
S3.
&i
Vo
7H
Tank Matnr
Tempgrature
totj.
/0a
ga.
sa
«-a
ffS
»a
s-/
ffCi
va
79
7«?
/y
ffo
79
So
Ambient
Temperature
SV
ff/
ffa
/?y
«a
»a
ga
SI
S>^3
»a
»a
5^
98
9^
4>3
9a
«?a
<« j
90
9/
*/
S>7
,54 '

-------
                                                             FURNACE OPERATING DATA
Cycle
No.









Cooling Water
Flowrate
(9P«i)
/'7









>










//










v











Thermostat
Sgttlns
£g









\










55










\j










,
Inlet Water
Temperature
-•T6.
^-£
57
i>~7
s(j>
£•<*
A't,
fT
A- 7
X7
fT7
SI
£7
.•S-7
&t>
A'C,
&(*
*£,
£<*
£7
S7
£7
«*
Outlet Water
Tempgrature
79
77
7fe
7 .V
73
7*4-
73
73
7*
71
7I
73
7«
83
a-.-v
sa
Vo
vz
(i*
1C,
C1$
/CO
103.
Tank Water
Temperature
IHa
//v-
//a
HO
;o&
/o 7
yoa"
/CiU-

/Ol
JOO
/o2.
/07
I/SL
117
/ja
/as
Xvl/
/..-?.-r
/3«?
/¥-"

(A)
ro

-------
                                                          FURNACE  OPERATING DATA
Cycle
No.


Cooling Hater
Flowrate
(9Pm)
1.7





















\




















Thermostat
Setting
l°C)
6V










































ti
Inlet Water
Temperature
4-4.
S7
&1
4L,
£-?
„*>"" £«•
^fc
6'(*
t>'7
3-C*
£3-7
S7
3-7
A' 7
d~6
5-6
A~ 7
.-5-6.
59
*7
.?«i.
y^-
83
6'«2
Sto
»?o
^S1
77
77
76,
7-5"
7^
7.3
73
>*
71
7/
Tank Hater
Temperature
/SI
/*-%
/V-G-
/*-/
/^tf
/3tT
/j?a
/as
A27
y^^t.
/^2/
/V9
/I 7
//s~
y/j?
///
/oV
/o/
/CJfc
/LJy.
/o3
/c^
/C'O
Exhaust Duct
Temperature
S13
6*3
»/
»/
&•/
ffo
77
Sfc»
ffc*
So
^0
£7
ffa
s*a
*a.
*3
ffa
fi'^Z.
tfa
^?a.
?a
sva
S'^
Ambient
Temperature
9^r
9
%3
S>7
ff-7
ff7
S»A~
fi»?
S>7
ffd,
6>»
6*7
g-7
ff6.
9$
89
97
97
?£>
#7
,96
CO
CO

-------
                                                           FURNACE OPERATING DATA
Cycle
No.
17







Cooling Mater
Flowrate
(9P-0
/7
'.7








































\f
Thermostat
Settinq
rO
fs
fsf









































-<7
.-s-
So
3
• S3
S3.
S*
83

-------
                                                          FURNACE OPERATING DATA
Cycle
No.
/*
Cooling Hater
Flowrate
(gpm)
1,7










V











//


















V
Thermostat
Setting
l°C)
£*










\
56










/


















v/
Inlet Mater
Temperature
57
_<6
A 7
S<~
£7
-•5*7
.56.
^-
Outlet Water
Temperature
6?/
TV
79
77
77
76
74*
73
73
rJ
•7.2
7/
73
78
JW
S1^
«?7
%)
«5-^
V^
£>6>
W
/oc>
Tank Hater
Temperature
/aa
//9
117
nS
H3
til
/o1/
/07
/ot>-~
ICI+.
/c-3
/oi
/oZ
IdS
1 13
US
I33>
/ax
IZI
/3k>
/3&
/^3>
/V-
-------
                                                             FURNACE OPERATING DATA
                                                                                                                               3ft
Cycle
No.

Cooling Mater
Flowrate
<9P»)
1.1





















\





















J
Thermostat
Setting
l°C)
S-5-





















V






















Inlet Hater
Temperature
°F
£<*.
£(»
S(*
*-*•
£(»
.<&'
£<-
3<*
art,
«*
s-t>
^•c*
s<*
trt.
£1*
A-tr
€<*
*"b
*L
f>i,
,<6
X&
£&
Outlet Mater
Tempgrature
10 /
/CM.
ioS
/o<*>
/Ot>~
/03
W
<*<*
V3
^/
iff
87
ȣ,
S3
«3
s-y
no
•n
TV
77
7t>
~7<~
7f-
Tank Hater
Temperature
»&
/SI
IS*
is -7
y
//4
1'+
//^
//o
Exhaust Duct
Tempgrature
S/
82
6>3
J?3
S>3
»3
»/
ffo
78
77
77
77
77
77
77
77
77
77
77
77
77
77
79
Ambient
Tempgrature
sa
ff/
So
ff3
<5to
9/
^a
90
S9
»«?
9/
85
ffC.
ffA-
ffi.
ff(p
tfa
#/
g-A
79
Sb
S>^
.®a
CO
(71

-------
                                                          FURNACE OPERATING DATA
Cycle
No.
ft
Cooling Mater
Flowrate
(gpm)
/.7

,

?
1.7




































V
Thermostat
Setting
l°C)
3-V


5S




















\,

















/
Inlet Mater
Tempgrature
£C
•$•<*
.5" 6.
£L,
S<»
S6,
S(*
&(,
S(»
A"•
Outlet Mater
Temperature
7*
73
"73
76.
S>
?(»
M

/*t-T
/*-a.
/3«
y.?:T
/?/
A3»
/J6
yjzy.
/^2J
//.?
y/7
Exhaust Duct
Tempgrature
78
r^'
/iff
S
?*-
fft
S>6

-------
                                                           FURNACE OPERATING DATA
Cycle
No.

Cooling Water
Flowrate
(go*)
/, 7

















i













r








Thermostat
Settinq
l°C)
*V
































1
V




Inlet Water
Tempgrature
6'C"
.< 6
5ft,
5' U,
.<6>
«"7
i>'7.
i."^
*'
*'(#
S7
<7
.6'C,
*."7
.i't-
4-7
.•s 7
A" 7
5" 7 '



3<^«M
Outlet Water
Tempgrature
77
7t-
7rf"
~7V-
7i+
7 a
73
7 a
73
71
70
7U
i,'i
fc'/
6*
&*
i.*
^
Ll



&&)
Tank Watfr
Tempgrature
X/rT
y/JJ.
///
/u9
>£)7
/oa"
/c>4*.
/o^
/oa.
yoo
y?
VSr>
5*6.
6/0-
9^
93
9a
9<
V/




Exhaust Duct
Temperature
7-iT
7*
-77
•75"
7.5*
7*.
7*+
7i>
7<~
7(a
-7C
7<-
7<°
76.
69
6t>
&>•?
£,«•
63




Ambient
Temperature
g-3
tfo
oa
S»o
83
79
-79
7?
7#
Tff
7*
77
77
77
7t»
75^
76.
7A"
70



,
CO
CO

-------
      SECTION 4
SAMPLING DATA SHEETS
      39

-------
                                                    HELD D«T* SHHT
        Plant
             ISr-S»DP.Jtffri
               HfcUrtF X.  (fOMrt^.>4t '
                                   0 0
                                               \)ft,
         Ur»e
    w,j^«-V
    » -{low
                                                               Ambient Temperature.
                                                               Barometric Pressure _
                                                               Static Pressure Duct _
                                                               Ftml   £><*T<^t\-
                                                                                               - 7S
                                                                                           Oil.
                                          DHY UNCOHRECTCO
                                              so,
                                                  NO,
                                                                 COHKCTtOTO
NO.  NO,
                                                                            HVOROCWWONS (HCI
                        a
                       Tout
                       MC
                                                                                         H,0
                                                y^
                                                	IT—*—
                                                * *ff
                                                      12-
                                l.f
                                                                                   ZLi
-p.
o
         A 111
                                i.as
                           n .1
                               X.,05
                                        ISO
                                        li.
                                                                                                      •f ^r'1*- t)a-t-
                               J.a
                                         SS

                                                 i£.
                                                                                                          OLU'^,  a.
                               t.Q
                               a.o
                                         30
                                                  7l
                               J-l
                                         30
ill
                                        150
                                                                            /. *
                                                                                                         t*"
         7/J7
      ±
                                         £7,3
67.
                                                      Z2_
                                    12.5
     J2_
                                                                            O. 9
                                       »s.e>
                                                                                             tfl.
                                                  tf
                                             I?
                                       fir
                                                  7o
                                                                                                          Form EECMM3

-------
                               FIELD DATA SHEET
Maother
                                                             - <97
Location
Operator
H^tirer (On,iw4l''>-'1i '•aU.y^'Kify
(frtJr^J).J,
5_^,. o.

tJi\tt-t Tu..
TDM
7:«H


SlO?
S\ 10
& : vi
fl : ik
& ! i3
£:i«4
D • 1^
B Mfc
<6; i7
^.^^


<#'V6
f3:»o
£:53
&3
73
fo
•jo
Jl
7Z.


66.S
£7
(S
£?
6?
f-J.S







NO,
ppmn
o


o
o


















o











p






CONNECTED TO
fJTIk O2 DB»
NO
ppm




























""m




























NO,
ppm




























metric Press
c Pressure I

>IIOl

^u
HYDROCARBONS (MCI
'«*
O-S


3
3
-?.6
Zo
1.7
«.5
M
l.z
O,«)
O.0


^.
-
^
_
—
-
—







MMIIIIK




























Noa-
M«MM
HC




























H,0




























c,f
•f
67J


6i.9
£P.7
7ft 4
76. f
7f.z
*a/
to??•. t
p-r.£







M«. ^
COMMENTt
/3wn*f J^/-


lT*rt •/ 'yt/e l^o •?


tixr^M 0-
-K'^. - Ify.^.


( '

K^KM^ /.ft-


£i,,t- «/£> Ai  Cj Ht
-------
                                                          FIELD DATA SHEET
                                                                                                                 UREX
                                                                                                              Aerothenn
Plant
     Kt&iSe.iTifti, OAT*a. 4Wirx Wi*ft H.AJ.  c>
          Oat*.
                                                                                 Ambient Temperature.
                                                                                 Barometric Pressure _
                                                                                 Static Pressure Duct _
                                                                                 Pmii   S)'' "^i I|?K
                                                                                                     73-
                   "View  «
                                                                                                    K)'
                                                                                                      /g
                                               DRY UNCOmiCCTEO
                                         CO,
                                         so,
                      NO,
                                                               CORHECTEO TO
                                                                 »O2DRV
                                                                                     HVOHOCMieONS (HOP
                                                                                     ToMI
                                                                                     HC*
H,0
                                                                                                                   *****
                                   H.07
                                                                                    5.1
                                   x.oz
                                                            67
                                                                                    5. -
                                   f.Ob
                                             6*7
                                       i-i
-p.
ro
                                   ,7.07
                                        in,
                                             6?.}
                                                                                      -3
          JLLJ
                                                       70. •/
                                     7
                                        /Z.I
                                                   70.6
                                   Jt.Ol
                              n.8
                iff-
                                        11. 9>
                                                             7/
          JO^OL
                               lit
M.
                                                             (0
                          z,i
                                              o.c
                                                             t,T,
                                             4Qo
                                                   I??
                                                            ^£_
                                        \1.0
                                             ao.r
 10: n
                                   Afii
                                         12.0
           IQJi
                                    4i£
                                                                                                                       Form EECMM3  \tl
           a  -

-------
                            FIELD DAT* SHEET
M«II N«s>uej|irti_ ~T'ie«. Tretut*- W.T
+ n.rt.M. >OURJT/I
Location "n^n.e.11 /
Operator
OlJTA«.L)i»li
5-^«- «?o
Ln«i a (^Ae/H i r»



Ambient Temper
Barometric Press
Static Pressure C

•lura /A- CSfe ' /VOOM 7^t**,3jl4t i&T/tvf' Q^ S^'SS'C vS'Vxuit- V^3. ^ /:Jl
ln-«M
IO-«t
/O.'-S?


//;//
//;/£
////7
//io
/// * /
j/.-ft
//••/J
// //•/
l/;jf
II ;U
//.-Z?










Potal
fK^Aal-
JX»|-

















































LOM
MW




























SMm
Flow
.1000




























MV UNCONNECTED
*
/•7rf"
/. 75
/ Zr
I.b7
/.<>"


-'.X
if.l
1.16
Jt.oi
I. IB
\ n
1.^ 1
I.Bfc
I ^j
l.f&
1.75"










CO,
/2,b
/i.(,
/J7
IM



/&£
12. b
ll.t
I/.7
13.0
l*-1
"•?
ft.l
13,2
\J.1
13. fc










CO
•ciR-
J?00
fa?
tf/3
*i.8
-T/t>


/6f
4f\
•










so,
mm




























NO
pom
££
(7
7O
69.7
7l


£J.1
h-\
6&b
-
..
^
-
-
-
-
_










NO.
opm
es
(7
70
69.7
72


(J.I
M
6f?.b
fit)
f0.h
7ft?
lo-tt
ft .8
72. i
7i.l
T/.o










NO,
ppmn
o














































*



CONHECTCO TO
NO
POD




























NO,




























NO,
ppm




























monocMwoNS IHCI
T»
-
/.£>
I.-J
f.o
-


—
4 1
fill
3,°
\'l
1.4
l.t
oA
o.3
rt.fc










MMMn.




























Non.
MMhMI
HC




























V




























SWk
OM
•F
H
77
^9
flt>1
Si A


66
7i.i
75.6
77, i
7?.t
fBt.1
&>•&
£'r&
Sl,t.
M2.I
82. 1










COMMENTS
ry«z*/* /o.a
•a.^^

i
So,^ »//
' '

Xf*** CfeJ<. iJo. ^
V

V^WMW On
^C l'ta*f • 1 Zf ^ ^|




y
StM^t. j*A.
' '






AVERAGE


. .« Form EED-063 12/7B
* AJ  CsHa

-------
                                               FIELD DATA SHEET
     KfJi£?
Location
Operator.
Load
                                                                     Ambient Temperature.

                                                                     Barometric Pressure _

                                                                     Static Pressure Duct _
         5-21-80
                                                                                        0.1
                                                                                           .   5"    8
                                    DRV UNCOMIECTED
                    MM
                    Id/hi
                         Z.t>
                               CO,
                             13.9
                                   CO

                                     -
                                   {(<-
                80,
                                                  65
                                                       NO,
                                                             CORRtCTEOTO
                                                                    NO,
                                                                          HTDBOC/UWONS (HCI
                                                                          HC*
                                                               H,0
                                                                  £L1
                                                                                                         COMMfNTS
                                                                                                 TS*s4- fid.  Lfa.to
 iilSfc
                         /.Ff
                                   iLi
                                                                                                       -t.-LT.  s
 tl'.ea
                                                                         02
                              /OZ
                                                                                                       -#-
                                                                                                            TJrrr
                              ^Ll
MI2SL
Mi
                              fr.*
                                                                                                               34.
                         L2i
                                                                          2.0
                                                                                                                15.
                                                  73
                              as.
                                                                                                            "^TTT
                         I.H*
                                  ISO
                                                 if.
                        Mi
                                                                                            /flb
                              11.
          i^
                                                                          VH
                                                                                           A
                                                                                            7*,?
                         ii£i
                                                                          J.o
                                                 2Li
                                                                                            ff.
                         JLZi
                         Lil
                               12,6
                                                                                           71- <
 13./
                             Jlii
                                                                               ^
                                                                                                           Form EEO-OS1 1Z

-------
                                                           FIELD DATA SHEET
                                                                                                                   Merotherm
              K^ogj"
_____

Inratlnn  Pl

Operator
         load Range_
                                  i., ft :> J
                                          Ambient Temperature.

                                          Barometric Pressure _

                                          Static Pressure Duct _
                                          Fiml    SX*
                                    A?*
                                                CRT UNCOfinECTEO
                              Flow
                              nn*
                              • 1000
                                 *%*
      CO

      T?*.
so,
MO,
                                                                 COHHCCTEO TO
                                                                     02 cmv
NO,
ppm
                                                                                        mOHOCMWONS 
           IV
                                          17.
                                    /.Yl
                                                                                                 76 ?
                                                                                                          A3. 1
          ll.'Si
                                Jia.
ai
                                    /.ift
                                                     70.1
                                              Ms
                                                    &i
          H't?
                                     47.3
                                                     fa.tr
                                                                               •*"•" -/St.  •«.'.
                                                              ^iA
          W-.il
i?lL
                                                              65
                                                                                                 7/.s-
                                                                                                                              JA.
                                                              70,1
                                                                                       '.6
                                                              72.0
                                                                                                          flJ.4
                                                                                                                          Form EED-OtS 12/T

-------
                                                           FIELD DATA SHEET
         Plant
                                  fa.^c^ Lj.7>t HftJ
                 rTCmr..<
                                                                                  Ambient Temperature.
                                                                                    ,.~ ..... 2,3.
         Optnio
                                                                                  Fuel.
                                                                                                                 OH
         Load Range.
      Z»^
                                   /v O-Sl
            O^»-Vx
TU.I  <«TO   •   M A?**
                                                                                                ^ Wv
                                               OBY UNCOdRCCTED
                                                    so,
                                                       CORRECTED TO

                                                    OK » O3 OKI
                                                              NO,
                                                                                      HYDROCARMMS IHCl
H,0
                                                                                                         TOT*

                                                                                                          •F
                                                                                                                      COMMEHTI
                UkiHlt
                                          tl-1
    1L
                                                                                                                    S^jti
          15 Ha
                          af.e
                                                                                      1,7
                                         124
          IS: <
                           li-7
                                                             7».o
-p.
en
                                             se
                                                                                                        S3.7
                                                                                                    -*
                                                                                                        (5?, 7
                                                                                                  feit.
                                        11.
                                          £6.2-
                                    .70
                      ft-Z-
                                                              l.t
                                                                                      VK*
                                             11,0
                                                                   •*!?
                                   '•?#
                                                                                                            -*•
                                              rg
                                             15-5
                                                                                           £id
                         J£_
                      li.fi.
                                                                                      S.I

                                                                                                               K.I.-...   j«.p
                                                                                                                        FwiriEEO-OU  IJ/rt

-------
                                                 FIELD DATA SHEET
 Ptint
      »\
Location.
OfMfltOf.
Date	
A^r
                                                                        Ambient Temperature.
                                                                        Barometric Pressure _
                                                                        Static Pressure Duct _
                                                                        Fuel	
~TU'i—
                                                                                                          it It. -fp OWt*^.
                                                                                                       a/a
                     Fkm
                     Wl*
                                      DRV UNCORRECTED
                                cot
                                                              comecTEO TO
                                                                isozonr
                                                                      NO,
                                                                            HYOBOCABBONS |HCI
                                                                            Tolll
                                                                            HC*
                                                                                   MMtiim
                                                                                     HC
                                                                                                           COMHCMTS
                          0.5
                                                                                                        frdk.  No n
                          as"
                                                        *•
                          0-
                            frl
                                 /
                               ituau
                                    f-*
               our
         V.I
               Otit,
                                  f-
                                                                                                           AVERAQE
                                                                                                                      TS/7

-------
Plant         fiCtUtffjf.  	  .   Barometric Pressure.

Dale	j£r./2^? in —~	Pitol Number	

SUrt Time	0(f&l	Meter Box Number.
                                      <3**7......
                                           .HaCL
                                                                PARTICULATE TEST  FIELD  DATA SHEET
Operator.
                              OrifirM r.oaitici«ii
>zzle Size & f>
jlecular Weig
wr>

£f~3\,fllw


lumber -/t4+9.a. 	
Ml

FILTER DATA











IMPINGER
VOLUMES
S2fO
B40
-CL—

T__
3ltJ.
t+e ,

SILICA
GEL
1
TIME
y-**
CO:
2S
O2
r».
CO
^
 SAMPLE
  POINT
CLOCK
 TIME
VELOCITY
  HEAD
                                  ORIFICE
                                  METER
                                 AH in. wg.
   GAS
  METER
VOLUME FT'
                                                               TEMPERATURES "F
                                             STACK
                                                     PROBE
                                                            IMPINGER
                                                           ORGANIC
                                                           MODULE
                                                                              OVEN
GAS METER
                                                                                      IN
                                                                                           OUT
PUMP
ACUUM
in Hg
                  olIO
                                    'J&.
                                   3+7. 831
                                      63
                                    3+7
                                              7S
                                            asi
                                                                            77
 JSL
          LZ&-
                          379/3
                                              78
                                                                                     77
4*
00
                           12S.
                                              £,<+•
                                                     ast
                                                                            70
                                                                           71,
                 j2o£_
                                   39g-3J
                                            «gg/
                  gpg
         3g^
          LZfi_
                                                                                           77
                  a/^r
                 IZi.
                                              71
                                                                                   77
37* 3
 o?/g>
                                    iZZ.
                                                                                                 a
                 p^^
                           • 74
                                             ATI
                                              JZZ.
                  O.OO
                           LZ2L
                         4-78
                                     8-SL
                                                                           80
                                                                                      17
                          -7g
                                                               J8&3.
                                                                     fffl
                                                                           80
                  Soo
                                               77
                                                                                  71
         &IQ
                           122.
                  fn><77
                                                                              SSI
                                                                                   fib
          7J-*
AVO/TOTAL
                                                                                                            Form EEO-021 I n

-------
                              <2*/ a
PARTICULATE TEST FIELD DATA SHEET
1

Test Location &A&CtG££.Jfl&f* £f
Smtff


&rop
Srttxr


'STOP
AVO/TOTAL
CLOCK
TIME
-71, It.

g/'V-

Q<,3
*fo< 3

#8.1
<&•2o.>7

IQ9.7
/3*-7
AW 3

VELOCITY
HEAD
4P in. wg.
S.io
£00
£00

SLoo
3oS
3of

S.DO
/9o


WO
I9o
I9O

3oo
rto



ORIFICE
METER
AH in. wg
''T8
'77
'7-7

'77
• 78
•77

,78
•77
•77

•7*
•77
.77

• 79
'77
•77

•77
GAS
METER
VOLUME FT'
Sarrsz
.*o
S-fS./
&t,*-t,7S
ft,a t,z?
g-rt.stf
see AS

•rwota
t,/o.ia
Los.si
63a-97a
£33-973

£,37.4.3.
6t,3,#olecular Weig
lumber 	
11

— —


FILTER DATA
NUMBER



TARE



FINAL WT





IMPINGER
VOLUMES












SILICA
GEL

TEMPERATURES 'F
STACK
^^
78
ffa

73
•70
S3

70

Pa

fof
•7S


b-5
79


74"
PROBE
37$
at*.
3S9

269

tfrfS

<3&?
3t,3
asi

3.70
3.58
3Su-

zi&
3.t,L
$t,9


IMPINGER





















ORGANIC
MODULE





















OVEN
a


GAS METER
IN
So
ffVr
93

8\
es
%

S3
ss-
9S-

Sf

9*
-
83
ft


8
OUT
SI
SI
SI

SI
6V
6-a

^a_j
&a.
99.

"5*5^
^y **
Q ^C
ffa

ffgt
«>a
&3

^

FtlMI
I/ACIA
in H|
TIME

>
IM
1
-3
3
3

3
3
3

3
3
,3

3
3
3

3
3
^3


CO? O? CO

«









































Jf££s^*a
€ /3*9
Form EED-0?! 1 79
vo

-------
en
o

^ ' uorporanon
Plant . . fteu/t£t
€
Dale &-QI-8O
otat
0134-
c&o


e9«<

Test Locatic
Run Numbe
^ — f
*
- . . . Baruinetnc Pressure . . £?9'8 ~
— . Static Pressure 	 _• 9f$ tsi
yjf«-!^SIa
. - Pro
S&X
f
H*c>

:k Pressure Je9-&ef
be Number 3' ff/OtSS.
3^> "AP"I t. LI L A^-^. 1 to
Nozzle Size & Number _A5" 	
Molecular Waighl
BWO

FILTER DATA
Slack Diameter inches. ** 	 Pnot Coelticient — NUMBER TARE
Duel Dimensions in. x in 	 Pitnl Numbar &lbr>6-
Start Time £*"•»/. M(rt«r Box Number SI
Operator 	 "fytkoWl 	 ,. Orilire Cmtticiani 3-J3&
SAMPLE
POINT
Sntff

Srof
.tnuT


*n>r
Sn*r


3T/Htt


yrdP
Qrppjff*


Qrof

AVO/TOTAL
CLOCK
TIME
O
^
a-f
la*
I7,.7
f*»7
£>/<7
6£,7
•71. •+


VELOCITY
HEAD
9.10
Ao4

Ai6
AGO
ae>6

&&
3*6
3to

Silo
Seo
JOO

200
3 0O
AIO

£10

ORIFICE
METER
AH in. wg.
3-9
3.9

k-0
4*0
+>e>

Zl'O
ff^fi
4-/O

U-'O
U-O
XMO

0*0
it-0
t+-O

+,£>

GAS
METER
VOLUME FT>
<(•»». J33
tffrf'OO
STS.ojA
£78^9
399-3
>»ao<4t
t>3?.o63
&39.0S3
fAo-3
AS/ -7
•70l.«.t*
7oi. u±6
-730.9
-rw3.
7t>l*'i7&
7LU../TS
7S£-7
&07.2
&St7-W
G-S.7.IH*

FINAL WT.





T FIELD DATA SHEET
IMPINGER
4oo
Sexj i


•rao
*S"=


SILICA
GEL

TEMPERATURES °F
STACK
A3
fS

in-
77
sa.

6a
79
ffg

. KllN
77
SA
as
£6,
T6,
SI

4,3-

PROBE
3ff6
371,

3SS
379
377

38*
37f
371,

U*>(~
377
37f
.380
390
38.3.
380

' tuV

IMPINGER





















ORGANIC
MODULE
SU-
*£

St.
Jf,
&7

rfV
6-tt-
$¥•

fif
£Tt,
&9
(*c
So
SI
&l

6»

OVEN
#01
397

399
397
397

39f
3?6"
39*

39?
397
397
397
399
397
396

tUSO

GAS METER
IN
76,
77

7f
79
86,

77
SI
ffa

Tf
814

79 •
&+
«
-------
*+
TEST FIELD DATA SHEET
rjatp f "&/ — fi? <
y
Test Location jft&pA&f .&*>?!
Run Number
Stack Diameter inches —
Duel Dimensions in. x in
$tt
ft

ss
ft


sa
«-/
6"^

f9
&(y
S-7

&3.

OVEN
397
39(,

«£>/
397
397

4MX,
39S
397

Uol
W)
XQ7

3^
398
3f(.

3?(,

GAS METER
IN
Sf
**!

ft

sea

Slf.
Su
9*

SI
fit-
91

S3.'
89
?3

S3

OUT
ff/
ga.

SI
#3
83-

£V
83
9*

S3
S3
eu

ffa
93
8f-

6*3


PUMI
VACUl

TIME

»
m
i
/4*
/V-

X3
t-\
X3

/a
/3
/3
A3

13

COa Oi CO

*,






















3f- lift.
77«*
OH*******.


















' ' ' ' Form EED-021 1 79

-------
         ^^Corporat
               ation
      Plant

      Date .

      Test Location

      Hun Number
                                                                        -TEST FIELD DATA SHEET
                     Barometric Pressure _

                     Static Pressure	

                         Pressure	

                     Probe Number	
     Slack Diameter inches _
     Duct Dimensions in. » in.

     Stirl Time	

     Operator	
                    . Pilot Coefficient _

                    . Pilot Number	
                    . Meter Bon Number.
                                 . Orifice Coefficient,
MZle Sue & r*
alecular Weigl
wo
lumber 	
ii

FILTER DATA
NUMBER



TARE



FINAL WT.








IMPII
VOLt





«3ER
JMES





SILICA
GEL


TIME


CO;

O2

CO

       SAMPLE
       POINT
 CLOCK
  TIME
                         VELOCITY
                          HEAD
ORIFICE
METER
  in. wg
   GAS
  METER
VOLUME FT>
                                                                   TEMPERATURES "F
                                                 STACK
                                                         PROBE
                                                                IMPINGER
                                                          ORGANIC
                                                          MODULE
                                                                                OVEN
               GAS METER
                                                                                        IN
                                                                                             OUT
PUMP
ACUUM
in MB
                                                  -r?
                   379
                                                           .V*
                                                                                    39(.
                                                          87
                                                                                                    t-3
               /^.g-3
                 t'O
                    3-L
                    37+
                                                                                 39L
                                                                          93
                                                                                                       X3
               &I-L
 J909
 I Si. I,
                                          Irro,a9*
                    J*2-
                    38y.
J2-
                                                                                        93
01
IN}
                       JSO_
                               *-o
                                    J2fe_
                                                                                 399
                                                                                      13
                          /go
                                                          3So
                                                           57
                                                                   39S
                               "Q
                                                      M-
                                                                          £3
                                                                                94-
                                                                                                       /O
                       /9o
                                                          379
                                                                         fo
                                                                   399
                                                                                                 SO-
                                                                      /O
JStoi 8
                                                          371
                                                           £•/
                                                                                            9°
                                                                      iO
          *"*r
                                                  -73
                                                                                S3
                                                                                                       10
                                                          379
                                                                   399
         M*-
                                                  ea
                                            3So
                                                                             So
                                                   399
                                                             /o
       Omul
                                     73
                                                                                           91
                                                               ea.
                                                  T7
                                                                   398
                                                                                           83
                                                                     jLQ.
                                       373.
                                            383
                                                                                    399
                                                           90
       Sro*
Xoi-o
                                         /39a 355
1639
       Zrsutf
AQI-Q
                         i3sa
                                                                                        SI
                                                                                                    yo
      AVO/TOTAL
                                                                                                              Form EED-021  1 79

-------
                                  *•
                                            HAHMUULAIg TEST FIELD DATA SHEET
Plant Wet*'** r
Dale ..J?ii/t*flK£^t
Test Location ^—t
Run Number

. .. . Barometric Pressure _. 	 	 	
Slsi* Pressure
, __ _..._ Probe Number
Nc
Me
BV
Stack Diameter inches p,|^| CoeHicient
Mud Dimaminna in « in Dum Mimkar
Start Time

Dpocnlnr Drill,-. Conlficient
SAMPLE
POINT


9rt>S
s*»xr


grof
Srn&r


Srof>









AVO/TOTAL
CLOCK
TIME
£010
Si ho
3/3>&>
3.1 3, t,

333. L
33 1, 1
a&i i

3-34 I*











VELOCITY
HEAD
AP in 113.
/Sf
tif.4

MO
/SO
i/y* ^C*

X^t*^T
J3&
IA^











ORIFICE
METER
AH in. wg
H--0
H--0

4..0
4.-0
&..0

4-0

4-0











GAS
METER
VOLUME FT'
/*to3>~r
Wr>
/pat,.3J*
/^•3L3JV-
S&.t57 * (*
At-*!?&f&*
j^qcjSH.
/u»99&*.
/svy 5
/S3S-3










loll 8i>u-

zzle Size
>lecular V
& Number
Veighl .



	




FILTER DATA
NUMBER TARE



FINAL WT.






IMPINGER
VOLUMES










SILICA
GEL

TEMPERATURES "F
STACK
71
g-y.

76
y*
ffa

-73
•77
SI










-7<*
PROBE
389-
set

308
<*.7K
37*

393
378
375











IMPINGER





















ORGANIC
MODULE
S~0
£-0

SS
SI


4-3
ffS
S3











OVEN
399
398

Vo/
399
39»

399
398
397











GAS METER
IN
fff
9f>

83,
^6
9*

83.
97
9u.










8
OUT

-------
       £\ ACUREX
       «T A Corporation
PARTICULATE TEST FIELD DATA SHEET

Plant
''C***?'*'-.- 	 	 Barometric Pressure &7G7
Date 	 S-I7-&0
13+7




Test LocalK
Run Numbe
Stack Diarru
Duct Dimen
Start Time .
>n/£t/ftW*
r-»IL/__,
	 Static Pressure ..
C* -J*)6t». «*«.;_ SlacK Pressure +
•SIS m H^O
elfTttf !>..&.- . Probe Number J$_' &i-AtS,
Nc
Me
BV
Her inches — nr._ _ . riloi i^oenicieni i
lions m. « in. Punl Numhar 1

	 Meter Box Number £!6-3
Operator 	 &st**/i*J 	 Qrilice Coellici.ni • ~JI £
SAMPLE
POINT
SrtHcr


&»#

gm*.r



Sr/i/er


&n>f
VO/TOTAL
CLOCK
TIME
o
£
/O
I3>0
J3.0
/ff.O
sn-o
&f>7
a*>7
3e»f
36'7
38>o
38.0
4*3- O
»9 O
SO. (a
So-L
ff't
(,!>•<•
te'Z

VELOCITY
HEAD
AP in. wg
/9o
/fito
18*

/9o
/ffo
/a*

193
/7€
**>£

180
SSG
J+£

/t.0
JifO
J+4


ORIFICE
METER
AH in. wg
/ <97
J<9f-
J.9M.

J.9S
S93
J 99.

/'9*
1.9*
/<9St

1-9+
J 93
j.tfc

/??<*•
/ 9V-
/•93


GAS
METER
VOLUME FT'
A.98-90*
7*1 £9
7ot,.3>
•7oS 3l,nS7
7*0 -IS
7*3 -91



>ule Sue S l>
Secular Weig
un
NUMBER



lumber ft*±

FILTER DATA
TARE



' &~7J *•(









IMPIN
VOLlll
103.
too
O
~r*'
3ER
HES^
u*
(1*
— "^
SILICA
GEL
3So]sx..8
TEMPERATURES °F
STACK
1,7
7t,
91

7>?
78
SSL

73
80
6>A

76
•79
9SL

re
77
8S.


PROBE
360
36>t
&t*o

HS3
a.-r*

Z6>8
at.7
SL73

3*1
374.
373

*78
27*+
27*


IMPINGER





















ORGANIC
MODULE





















OVEN





















GAS METER
IN
fft
g,
SI

8t
8l
8/

80
3°
SI

&{
Si
e/
.
8/
Si
91


OUT
81
91


81
81
ff/

9a
81
81

&
3f
ffl

SJ
So



TIME

PUMP
VACUUM
hi Hg
S
,f
if

£•
g'
£

6-
5-
&

£•
f
,5-

£•
g
£-


CO: Oi CO

^





















zy~





















tn
                                                                                Form EED-021 1

-------
     ^^Corporation
PARTICULATE TEST FIELD DATA SHEET
Plant. -/VCHtf
Dale £-/7-&
€
9
Test Location ^UffUt^Lj^f^.-
	 Bare
" SIM
^5t? Stac
>melric Pressure..
c Pressure .
k Pressure 	 	
frft1Ttr-\
Run Number — . . Probe Number


	

Nozzle Size A f>
Molecular Weigl
lumber
ii







FILTER DATA
Stack Diameter inchex PJM Coefficient _NUMBEn, .
Duct Dimensions in. * in
Start Tim.
P»nl N,,mh«
U.l.r Rn» Mumhw
Dpantor nrifir. rnaHielMil 	
SAMPLE
. POINT
Gnwf


Sfof



<9raC












AVG/TOTAL
CLOCK
TIME
fcg.Q

T*<
-rt,4
*?£,.£

86- £
97.4













VELOCITY
HEAD
AP in. wg.
!•*£
/3f
ia*

I3o
Ja*
/•3f














ORIFICE
METER
&H in. wg.
/ 9^r
t'9tf-


htV.
/•9+
/•&













I&U
GAS
METER
VOLUME FT'
~-rjf.g-$l£

-7S3 itS
"yc-TiSHt-f
757«a*9
7S936

763-9X3












6>£-Otl
TARE



FINAL WT.






IMPINGER
VOLUMES












SILICA
GEL

TEMPERATURES °F
STACK
7H-
77
Si

77
c?0
SA













-78
PROBE
arts '
371
S.73

ati
2&?
3t,7














IMPINGER





















ORGANIC
MODULE





















OVEN





















GAS METER
IN
m
*l
91

si
Zt
s>/








•




0
OUT
c?/
^f/
91

5V
J?/
8/













/


MCUl
in H|

TIME

»
)M
i
5"
5"
5-

^>
^5-
5"















CO? O? CO

*





























fiA/B^W-C
/g»T«f -OI&













en
01
  has

-------
           SECTION 5



ANALYTICAL LABORATORY RESULTS
           56

-------
5.1    FUEL ANALYSIS REPORT
                                   57

-------
                               LABORATORY CERTIFICATE
                          _
                 CURTIS & TOM PKINS, LTD.
         orrci*k
   ANO'Oo tAMPiiat re* MANY
COMMODITY AND T»API e»8ANUATlQNl
                 ANALYTICAL. CH EM I SXS-CONSULTING
                              •AMPLER* —INSPECTOR*
                                    t»O DIVISION »THICT
                                 •AN rHANCIBCO. CALIF. B41O3
                                         U.S.A.
                                                                   •«•!.»•£•
                                                                 — IMVMTIftATIOMS
                                                          VITAMIN *•»*¥• - •lOCMIMIfTIlT

                                                         •VCCWUSTS IN BULK COMMOOITKI
' Laboratory No.  SOklO
 Preliminary No. 4132
 For
      ACUREX CORPORATION
                                                         Reported  10/6/80
                                                         Sampled   	
                                                         Received  9/8/80
 Report on  5 8an,pies of Oil
      \ ' ,
                       Sample #    Sample Type
         Mark         (5)  80-1562,     Fuel Oil
                      (All) Project No.:  7601.22, Customer No.: RB  59186A,  rel.  01,
                       Prime  Contract: 68-02,  3188, Acurex Project:  7601, Date:
                       September 5, 1980,  Subcontract No.: RB59186A.
                                                          (5)
Carbon
Hydrogen (H), T. —
Oxygen (0), by
 difference,?.	
Nitrogen (N) ,7. —
Sulfur (S),% 	
Heating Value:
 BTU/Pound —	
Gravity, •
 °API @ 60°F	

*Less Than
                                                   1st      2nd
                                                   Test     Test
                                                    86.94   	
                                                    12.90   	
                                                19,190

                                                    33.75
                                                         3rd
                                                         Test
                                                     0.01*  	
                                                     0.03   0.04   0.05
                                                     0.18   0.20   0.20
                                              58

-------
5.2    PARTICULATE GRAVIMETRIC ANALYSES AND MASS EMISSION CALCULATIONS
                                    59

-------
/•S ACUREX
T >
Corporation           PARTICULATE CALCULATIONS


             Too.  \\lfr* VW*ME?  ^T*CK  SAMPLE  ' hpTTHoO  5

    1. Volume of dry gas sampled at standard conditions, 68° F, 29.92 inch Hg (set)
                     Vm /  Pb * 13.6

              r 1764-irlT	^460"
                        llm,avg
                     i    + v«i  "\ /Ts,avg + 460\pn_\   (acf at stack conditions)
        or Vmstack' \~5~  vwstd/\T         ~" "   '
    2. Stack gas moisture condensed at standard conditions (scf)



        Vwstd = 0.04707 Vlc
    3. Stack gas proportion of water vapor, by volume
         o
         B
          wo - Vwstd * Vmstd
     4. Stack gas dry molecular weight (Ib/lb-mole)



         MWd = 0.44  (%CO2) + 0.32 (%O2) + 0.28  (%N2 + %CO)
     5. Stack gas molecular weight (Ib/lb-mole)


         MWS = MWd (1 - Bwo) + 18 (Bwo)
     6. Pressure stack, in. Hg
          s - rb T 13.6
                                         60

-------
7. Stack gas velocity at stack conditions (ft/sec)

                                                                   tft

     V  -B549(C                         46°
      s-    •
                                /Ts,avB * 46
                               ^   psMW5
8. Stack gas volume at standard conditions (scfm)



     Q8 . 60 (1 - Bwo) Vsavg A8  (



     or Qa = 60VsavoAs(acfm)=
          8        avg"s»



 9. Test percent isokinetic
                       -
       s     '    Vmstd "           1S6 .4


                       MD
     or  Cfl = 15.432 -n— -^ — (gr/acf) =
                    vmstack
11. Emission rate of participate matter. Ib/hr


     ER = 0.00857 (Qs) Cs
12.  Percent excess air at sampling point


                  100 [%02 - 0.5
           = 0.264 %N2 - (%02 - 0.5 x %CO)




13. Emission rate of paniculate matter, lb/106 Btu



     E = 2.679 x 10*
                    PL.W
                    VVmstd/   \
                    VVmstd/  \20-9 - %02
          17-33 (T5[8vg ^ 460)  [0.04707 (W|c) * Vmstdl

                         • vs ps Dn2
10. Participate matter concentration, gr/scf
           15 432
            '
                                     61

-------
                    PART1CULATE CALCULATIONS





 1. Volume of dry gas sampled at standard conditions, 68° F, 29.92 inch Hg (scf)
     Vmstd
                        m.avg
     or Vmstack
 2. Stack gas moisture condensed at standard conditions (scf)




     Vwstd = 0.04707 Vlc



           =  ^l.^ll





 3. Stack gas proportion of water vapor, by volume
                   .2C.Vf 3\,3M



 4. Stack gas dry molecular weight (Ib/lb-mole)




    MWd = 0.44 (%C02) + 0.32 (%O2) + 0.28  (%N2 + %CO)
5. Stack gas molecular weight (Ib/lb-mole)




    MWS = MWd (1 - Bwo) + 18 (Bwo)




         =   JO-iH (l-3.o)-»-l8(l..o)




6. Pressure stack, in. Hg
                                  62

-------
  7. Stack gas velocity at stack conditions (ft/sec)        .,    .,        .
                                                    VtUa^j  uss. 4.  j-^jit  -k  i*

      Vs = 85.49 (CD)              T     *          °
                  P
 8. Stack gas volume at standard conditions (scfm)
                         D
      or  C_ = 15.432 T,—-E— (gr/acf)
                    vmstack
11. Emission rate of paniculate matter, Ib/hr

     ER = 0.00857 (Qs) Cs
                                         r   o.
12. Percent excess air at sampling point
                                          --   l-
                  100 [%O2 - 0.5 (%CO)]
      % ^ = 0.264  %N2 - (%O2 - 0.5 x %CO)
13. Emission rate of paniculate matter, lb/10" Btu


     E = 2.679x10*'—K  ' = '    ?P9
                     VrnStd/   \20.9-%02
                            avg      PSMWS        ^Siow^t,, .  s^acfc  -YI   ^fcloe.'-fj
      or Qa = 60 Vsavg AS (acfm) =   60 (i-0.oi)^ £*   f*<3    £i£?  -  I fe. S '

  9. Test percent isokinetic

      ,,. .  17.33 (Ts.avg ^ 460)  [0.04707 (Wlc) + Vn^J
                          fl~V—P—D~»~	
                             s  s   n

         =    \v? %

 10. Paniculate matter concentration, gr/scf

      Cc = 15.432 -n-*- =    It
                                                  63

-------
5.3    SULFUR ANALYSIS REPORT AND EMISSION CALCULATION
                                 64

-------
                                 Mist and SO2
                           Clean-Up and Analysis
Plant
                                           Oate
         5".
Project No.
                     760$ '
Recorded by























H23C>4 Mist and SC>2 Train Description
Impinger Sequence 1 2 3
Impinger Number
G-S Standard ( )
G-S Modified ( )
Contents *?/) &0i- MA
Concentration ("'•} _£p ^ ^
Initial Volume vO /ft? /&/ 2 Anal
Sample Number Aft£ /// &0t/
Ttt. Vol. of Soln. (ml) 2$o £5o
Volume of Sample .
Aliquot Titrated (ml) *7*? *>o
Performed by (V'''4^»«. vOu&ru*

and Recovery Data
456



- ^^ ^- 	
e jlso.oji '•
3 jl?6.iK>
J

** Mt

•* [
6*. 07 \

>rSis
1







                                     65

-------
5.4    TRACE ELEMENT ANALYSIS REPORT
                                    66

-------
          COMMERCIAL TESTING  & ENGINEERING CO.
          MNltAL OFFieCt- IM HOtTH It, SAUt STRUT, CHICMO. IU.IDOI* W*0i • MI* COPt (It TM-M14
••ply 10     MS1IUMINTM AMMTMS MVWON. MM WBT MIN AVtNUf. OOUKN. COUMAOO «MO). MONt; MMM4S11
T* Mr. Brent Higgln
Acurex Corporati
485 Clyde Ave.
Mountain View, C
f. O. NO.:
Simpto No.: 80-1535
ELEMENT CONC.
Uranium
Thorium
Bismuth <0.1
Lead 0.6
Thallium
Mercury NR
Gold
Platinum
IHdium
Osmium
Rhenium
Tungsten
Tantalum <0.6
Hafnium
Lutetium
Ytterbium
Thulium
Erbium
Holnlum
Dysprosium
tat - Not ((ported
Alt MWIMRtl llOf OVtt0BO^
ibotham ^\
on ^S
A 94042
•WWK SOURCE MASS IPB
CONCENTRATION
ELEMENT CONC.
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium 0.2
Cerium 0.9
Lanthanum 2
Barium 1
Cesium
Iodine <0.1
Tellurium
Antimony 0.2
Tin 0.1
Indium STD
Cadmium
Silver
Palladium
Rhodium
* Heterogeneous
c 0.1 ppm
k
IN PPM WEIGHT
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Geranium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Appro*): 'I/
m
CONC.

0.4

1
<0.1
0.2
<0.1
0.4
0.2
<0.1
0.2
0.1
6
13
MC
3
120
39
29
M -
Me/
°»* September 23, 1980
Analyst: T. Bouts
IAD No.: 97-E697-116-28
ELEMENT
•Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
N1 trogen
Carbon
Boron
Beryllium
Lithium
Hydrogen
%«£-*n*-
CONC
0.3
2
<0.1
170
24
1
9
26
*MC
13
17
98
=3
NR
NR
NR
<0.1

0.1
NR
£
  MC - Mtjer Compenwtt
  •UT
  mi ^
                                                  67

-------
Reply to
COMMERCIAL TESTING & ENGINEERING CO.
eiNEIAL OmClt  IM MOUTH LA IALLC «T*IET, CHICAGO. ILLI«OI» IMOI  • A»f A COOC «1S T»»-»4»4
  IMTIUMNTAl ANAmil tXVIVOH, IOU WIST MTH AVfNUE. 6OIMM. CtXOtADO IMOI. fMONi: 103-1714991
T»=   Mr. Brent H1gg1nbotham
     Acurex Corporation
     485 Clyde Avenue
     Mountain View,  California 94042
                                                                  September 26, 1980
                                                                               T. Bouts
P. O. No.:
Sempte No.: M75 5pARK SOURCE MASS SPEaROGRAPHIC ANALYSIS IAD No- 97-E697-116-28
l^.TW.'Ur.i "&».ft*ite CONCENTRATION IN vg/ml
ELEMENT
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridlum
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutetium
Ytterbium
Thulium
Erbium
Holmium
Dysprosium
CONC. ELEMENT CONC.
Terbium
Gadolinium
Europium
<0.02 Samarium
Neodymi urn
NR Praseodymium
Cerium
Lanthanum
Barium * 1
Cesium
Iodine 0.01
Tellurium
Antimony
Tin 0.2
Indium STD
Cadmium
Silver 0.5
Palladium
Rhodium

ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium

CONC. ELEMENT
Vanadium
0.05 Titanium
Scandium
0.01 Calcium
Potassium
0.006 Chlorine
Sulfur
0.02 Phosphorus
Silicon
Aluminum
Magnesium
Sodium
0.07 Fluorine
0.07 oxygen
0-03 Nitrogen
0.004 carbon
0.09 Boron
0.006 Beryllium
0.006 uthlum
Hydrogen
CONC
0.002
0.01
0.001
6
2
0.3
0.2
3
0.4
0.3
0.08
* 6
=0.1
NR
NR
NR
0.001

0.002
NR
™" ^ ww Reported
AH elements not detected^
•wt ••• Me|or Component   ',
INT — InlerfeieiiLe
                       0.004
                    g/ml
                    i
                                              Approved:
                                                 63

-------
••ply*
COMMERCIAL TESTING  & ENGINEERING  CO.
MNMAL OFFICO: tit NOKTM LA IAU.C »T»MT. CHICAGO. ILLINOIS MUi • AMA CODC HI TM-MM
  MSTMMINTAl ANAiniS MVWON,  1AM WOT MIM AVINUf. GOIHN. COtOIAOO «M01. MtONt; >QH>MSi1
To: Mr. Brent Higginbotham ^Bk
Acurex Corporation ^*5flS^^
485 Clyde Ave.
Mountain View. CA 94042
P. 0. NO.:
$wnpi« NO.: A-45 Blank SPARK SOURCE MASS SPECTROGRAPHIC ANALYSIS
D*** September 25. 1980
Ai»ly«f: T. BOUtS
IAD NO.: 97-E697-116-28
^n.Tc 10 vg/cm2
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium


Approved; /
>
CONC.

0.005
0.01
0.05
0.003
0.04
<0.001
0.03
0.005
0.001

0.008
0.01
0.01
0.01
< 0.001
0.2
0.003
0.02

st // /
VI 9
i
ELEMENT
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Hydrogen
'#/
'JUg^&fl

CONC
0.02
0.9
0.002
MC
0.5
0.4
0.8
0.07
MC
MC
1
>2
•0.005
NR
NR
NR
0.9

<0.001
NR
tf
-~fa
                                                  69

-------
          COMMERCIAL TESTING  & ENGINEERING CO.
          •cmMI orrict»: in HORTH i* *ALLI trncT, CNICABO, lumoi* M«OI • *«• eoei nt ?t»-««M
••ply tO      MKIUWN1M MttlVSU WVMION.  1AM WIST *OH AVtNUI. OOUXN. COtOMOO tO»l. MOM: JBM7MSJI
To:   Mr.  Brent Hlgglnbotham
     Acurex Corporation
     485  Clyde Ave.
     Mountain View.  CA  94042
                                        V*llL.
                                       mmrnmm.
 P. O. No.:

 StmpteNo.:  80-1312

TAP DATcf, i
                       SPARK SOURCE MASS SPEaROGRAPHIC ANALYSIS

                             CONCENTRATION IN  jig/ml
                                                                        September 24,  1980
                                                                  An«iy*t:L.  Jacobs
                                                                  IAD No.:  97-E697-116-28
ELEMENT
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutetian
Ytterbium
Thulium
Erbium
Holmlum
Dysprosium
STD - Mm
NK-Noth
INT «« hitarfi
CONC. ELEMENT CONC.
Terbium
Gadolinium
Europium
0.01 Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium 0.02
Cesium
Iodine
Tellurium
Antimony
Tin
Indium STD
Cadmium
Silver
Palladium
Rhodium
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
21 nc
Copper
Nickel
CONC.

0.1
0.005
0.02

0.02
10.002
0.1

0.004

0.009
0.04
0.002
0.003
Cobalt *<0.002
Iron
Manganese
Chromium
0.2
0.006
0.07
«l Snnctant * Meter°9en«>US
^^>io°d/SW Appw-: ''1M ^
tranc*
r
l( (/A
ELEMENT
Vanadium
Titanium
Scandi urn
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Hydrogen
//
ft^e-i
CONC.
<0.001
0.1

10
0.05
MC
0.4
0.9
0.2
0.004
MC
0.2
"0.004
NR
NR
NR
<0.001

INT
NR
&
                                              70

-------
 ••ply to
COMMERCIAL TESTING &  ENGINEERING  CO.
•MCtAL OmCM: ttt W*TN IACAUI »TIKT, CHICAGO. IU.IMOII IMOi • AM* CODE ttt »•••«•«
  •muMMTAi ANMTSn MVWON.  MM WOT 4«IH AVWUf. SOIMN. COlOtAOO M«l. tHOHt
 To:  Mr. Brent Hlgglnbotham
     Acurex Corporation
     485 Clyde Ave.
     Mountain View. CA  94042
 9. O. No.:

 SwnphNo.: 80-1313

WftTcft T
              SPARK SOURCE MASS SPECTROGRAPHIC ANALYSIS

                    CONCENTRATION IN
                                                            : September 24. 1980


                                                         A««iytt:T. gouts


                                                         IAD No.: 97-E697-116-28
ELEMENT CONC.
Uranium
Thorium
Bismuth
Lead 0.03
Thallium
Mercury NR
Gold
Platinum
IHdium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutetlum
Ytterbium
Thulium
Erbium
Holmlum
Oysproslum
STD - hitaffwl $».nd»rd
* * NR ^ Not Rcpofftd
All •!• • •nil m*ut j^t^*^^
All •••menu nor avi^cwn
MC- M»jor Compoowrt
ELEMENT CONC.
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium 0.2
Cesium
Iodine °-01
Tellurium
Antimony
Tin
Indium STD
Cadmium
Silver
Palladium
Rhodium
* Heterogeneous
i<0.003 v9/ml
> 10 vg/ml
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Z1nc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Approved: /
CONC.

*0.1

0.02

0.03
0.006
0.3

0.002

<0.007
*2
4
0.04
0.005
0.3
0.01
0.008
WL
ELEMENT
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Hydrogen
/A~
CONC
0.003
0.1
<0.001
10
2
8
2
0.4
2
0.2
0.5
2
•0.1
NR
NR
NR
0.002

0.004
NR
-A
   •UT
   mi ^
                                                 71

-------
           COMMERCIAL TESTING &  ENGINEERING  CO.
           •iNt»i ernctt: tt* HO»TM u «»u.« §T«HT, CHICACO. IU.IMOI* MMI • »•§• coot >tt >*«.M*«
R>ply 10     MHMMNTM. AHAtTSII WVIilON.  14BS WOT 44TH AVINUI. 6OIMN. CMOMOO Ml. »MOtW M-XTMBI
T<*   Mr. Brent H1gg1nbotham
     Acurex Corporation
     485 Clyde Ave.
     Mountain View, CA  94042
f. O. NO.:

Swnpte No.: 80-1368
                        SPARK SOURCE MASS SPECTROGRAPHIC ANALYSIS
o.t«: September 24,  1980


An«ly»t T. BOUtS


IAD No,: 97-E697-116-28
-—. — _,_ bunwENiKAimn IN Ma/1"1
ELEMENT
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Irldlum
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutetian
Ytterbium
Thulium
Erbium
Holmium
Dysprosium
STD- Men
m«j— •*.
•"* NO? mt
All^tonwnta
MT-kMrft
CONC. ELEMENT CONC.
Terbium
Gadolinium
Europium
0.1 Samarium
Neodymium
NR Praseodymium
Cerium 0.008
Lanthanum 0.01
Barium 0.07
Cesium
Iodine
0.02 Tellurium 0.02
Antimony
Tin
Indium STD
Cadmium
Silver 0.03
Palladium
Rhodium
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Z1nc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
CONC.

*0.4

0.05
0.05
0.05
<0.001
0.4
0.1
INT
0.002
0.004
MC
MC
1
0.02
MC
0.2
0.7
* Heterogeneous
wl Standard
•W **cMd< 0.002 ng/ml
JJJ2"""1* > 10 ug/ml
Approved: -'y
?n
ELEMENT
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine *
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Hydrogen
/ /

-------
          COMMERCIAL TESTING  & ENGINEERING CO.
          •ENMAL OFFICE*: »• NORTH LA «AklE (TVEET, CHICAGO, ILLINOIS «M01 •  AKA CODE Itl Tl«-«4»4
Reply to      msnuMiNTAi ANAITSU DIVISIOM.  MBS WEST MTH AVINUE. GOUKN. COIMAOO «M». FHONE: aa-i7**si
To:  Mr.  Brent H1gg1nbotham
    Acurex Corporation
    485  Clyde Avenue
    Mountain View,  California  94042
                                                                      o*te: September 26. 1980
                                                                      An«h/*T: T.  BOUtS
9. O. NO.:
Ssmpte No.: 1471
HArj >t«.r
~^f \\-~f_tL
ELEMENT CONC.
Uranium
Thorium
Bismuth
Lead 0.2
Thallium
Mercury NR
Gold
Platinum
Indium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutetium
Ytterbium
Thulium
Erbium
Hoi ml urn

SPARK SOURCE MASS SPECTROGRAPHIC ANALYSIS
CONCENTRATION IN v9/cm2
ELEMENT CONC.
Terbium
Gadolinium
Europium
Samarium
Neodymium 0.005
Praseodymium 0.003
Cerium 0.03
Lanthanum 0.04
Barium * 0.9
Cesium
Iodine
Tellurium
Antimony
Tin 0-02
Indium STD
Cadmium
Silver °-002
Palladium
Rhodium

ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium

CONC.

0.05
0.005
0.006
0.008
0.08
<0.001
0.04

<0.001
<0.006
0.009
0.6
0.3
0.04
0.02
0.7
0.02
0.1

IAD No.: 97-E697-116-28
ELEMENT
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Ni trogen
Carbon
Boron
Beryllium
Lithium
U \fft *tAM API
CONC
0.2
1
<0.001
MC
0.5
0.3
8
0.5
MC
MC
4
MC
=1
NR
NR
NR
MC

0.02
NR
  (TO - Intonwl Standard
  NR - Not Iteportod                   2
  All •tenwnti not dMMMd< 0.001 UO/Cm
  NIC - Mijor Component   >10ug/Cffl
  IUT   t
  mi ^
                                                                       * Heterogeneous
                                                 73

-------
Reply to
          COMMERCIAL TESTING & ENGINEERING  CO.
          •tNCML orriet t: it* NORTH t» «*ut •TMIT, CMIC»«O, ILLINOIS Mtoi  • »«» CODE tti ?f t-»4i«
            msnuMiNTM MUITSIS DIVISION. i«tt WIST MTM AVINUE. GOiofN. COIMADO «woi. mowf, JOJ-ITMSJI
To:   Mr.  Brent HIgglnbotham
     Acurex Corporation
     485  Clyde Avenue
     Mountain View, California  94042
                                                                             Septenter  26, 1980
                                                                      Arwlyit:
f. O. No.:
Simple No.:
ELEMENT
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Irldlum
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutefium
Ytterbium
Thulium
Erbium
Holmlum
Dysprosium
sm _ *— * 	
1476 SPARK SOURCE MASS SPECTROGRAPHIC ANALYSIS
¥ OKC, CONCENTRATION IN vg/ml
CONC. ELEMENT CONC.
Terbium
Gadolinium
Europium
0.04 Samarium
Neodymlum
NR Praseodymium
Cerium <0.001
Lanthanum
Barium 0.05
Cesium <0.001
Iodine
Tellurium 0.01
<0.002 Antimony
Tin 0.05
Indium STD
Cadmium
Silver 0.03
Palladium
Rhodium
t e*«^___i
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium

CONC.

0.008

0.002

0.002
<0.001
0.004
0.006


0.002
0.2
0.2
0.2
0.004
0.4
0.04
0.08

IAD NO.: 97-E697-116-28
ELEMENT
Vanadium
Titanium
Scandi urn
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Hydrogen
CONC.
0.001
0.3
<0.001
1
3
0.04
MC
3
1
>0.7
0.6
MC
=0.3
NR
NR
NR
0.003

0.005
NR
 NR - No* *(porl*d
 AII •iwMnh not d>i»cf*d< 0.001  vg/ml
 MC - Mijer Component  >1Q
 INT — bntrftrcna
                                             Approved:
                                               74

-------
           COMMERCIAL TESTING & ENGINEERING  CO.
           •EMIIAL OPFICM: t»i NOVTH Uk IAU.C tTKET, CMICAOO, ILLINOIS «0«OI •  »«» CODC 111 T»l-*4»4
••ply tO     INtnuMNTM. AMAiniS DIVISION.  I«UJ «WST «TH AVfNlrt. GOIDCN. COlOtADO KM01. PHONE: 10.17*4531
To: Mr.  Brent Higginbotham
   Acurex Corporation
   485  Clyde Avenue
   Mountain View,  California   94042
P. O. NO.:
September 26, 1980
                                                                       *nily*t:
Simple No.^ i35g spARK SOURCE MASS SPECTROGRAPHIC ANALYSIS
TA *~-'-' T* ' - A— -- - CONCENTRATION IN vg/ml
ELEMENT CONC.
Uranium
Thorium
Bismuth
Lead 0.09
Thallium
Mercury NR
Gold
Platinum
Indium
Osmium
Rhenium
Tungsten
Tantalum <0.04
Hafnium
Lutetium
Ytterbium
Thulium
Erbium
Hqlmium
Dysprosium
$TO - IM*m«l Standard
NR - Not Reported
All wwwnti not otTtctvo ^
MC - Mijor Component •
ELEMENT CONC.
Terbium
Gadolinium
Europium
Samari um
Neodymi um
Praseodymium
Cerium
Lanthanum
Barium 0.02
Cesium
Iodine 0.004
Tellurium 0.006
Antimony 0.02
Tin <0.006
Indium STD
Cadmium 0.006
Silver
Palladium
Rhodium

0.002 pg/ml
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium

CONC.

0.2

0.03
<0.2
0.009

0.1
0.1
<0.009

0.002
MC
MC
* 0.7
* 0.07
7
0.1
0.5

IAD No.: gy.
ELEMENT
Vanadium
Titanium
Scandi um
Calcium
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodi um
Fluorine
Oxygen
Ni trogen
Carbon
Boron
Beryllium
Lithium
Hydrogen
•E697-116-28
CONC
0.002
0.2
0.01
MC
3
* MC
MC
0.8
6
0.7
2
2
«0.2
NR
NR
NR
* 0.03

0.003
NR
f Heterogeneous /?*.
t
w, c
f if -^«-
^ "
                                                   75

-------
••ply le
COMMERCIAL TESTING  & ENGINEERING  CO.
•fNt*AL OrriCtt: tte MOtTH LA MU.C tTKCT, CHICAGO. lUIHOIt eoeot • AMA COOi MI fl»-»4»4
          , ANAinu WVIIION.  uns urtji *m» ATOM. oou*H coiotAoo *MOI. moNi: jowrwai
T«* Mr. Brent H1ggi
Acurex Corporat
485 Clyde Ave.
Mountain View.
P. O. No.:
Simple NO.: 80-1473
yA£> ~ MrtrJ TEST

ELEMENT CONC.
Uranium
Thorium
Bismuth
Lead 0.3
Thallium
Mercury NR
Gold
Platinum
Iridlum
Osmium
Rhenium
Tungsten
Tantalum 0.7
Hafnium
Lutetian
Ytterbium
Thulium
Erbium
Holmlum
Dysprosium
fTO - Internal Standard
N« - Not Reported
All •tonwnti not delected <
MC - Meter Comment
nbotham J
1on ^"
CA 94042
••ARK SOURCE MASS W

ft
•ECTBOOHAPrtC ANALYSIS
D.I. September 23, 1980
*n«iy»t: T. Bouts
IAD No, 97-E697-116-28
CONCENTRATION IN PPM WEIGHT
ELEMENT CONC.
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium 0.3
Lanthanum 0.4
Barium 0.9
Cesium
Iodine
Tellurium
Antimony
Tin
Indium STO
Cadmium
Silver
Palladium
Rhodium
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
CONC.

1

<0.1

0.2

0.8
<0.1
<0.1

*<0.1
6
6
180
0.7
26
1
11
* Heterogeneous
* t

0.1 ppm

/}/

ri|f|iiuv*Mj. u A
I/I ( /

'/, 1 / A
ELEMENT
Vanadium
Titanium
Scandi urn
Calcium
Potassium _
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Uyd/bgen
y/
r* ^
L£~^**-*
CONC
<0.1
*2
*<0.1
62
22
1
14
15
*500
5
8
22
**0.3
NR
NR
NR
<0.1

<0.1
NR
s?
&i
7A.
BAIT —
•«* ^
                                               76

-------
toply to
COMMERCIAL TESTING &  ENGINEERING CO.
•ENttAL OFFICES: SSS NO«TM LASALLE STKET, CHICAGO. ILLINOIS SMOI  • AIEA CODE S1I TM-MI4
  WSnUMCNTM ANAirSH DIVISION.  MM WIST «TM AVEMUt. COiBCN. COIOIADO SMOt. PHONE* M*-«S«S9I
To: Mr. Brent H1gg1
Acurex Corporal
485 Clyde Ave.
Mountain View,
f. O. No.:
tempi* No.: 80-1562
ELEMENT CONC.
Uranium
Thorium
Bismuth
Lead 0.2
Thallium
Mercury
Gold
Platinum
Irldium
Osmium
Rhenium
Tungsten
Tan tali urn
Hafnium
Lutetian
Ytterbium
Thulium
Erbium
Hplmlum
Dysprosium
iTD - M*rn*l StwidMd
NR *•> Not teportvd
All •!••!• Ill* BkM* JaW^M^M^
mi vwvnvnT* n*7* OTI^O^V
MC - Major Component

nbotham ^i
:1on "•.
CA 94042
__ SPARK SOURCE MASS SPEC1
|T '»' CONCENTRATION 1
ELEMENT CONC.
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium 0.3
Cesium
Iodine
Tellurium 0.2
Antimony
Tin 0.03
Indium STD
Cadmium 0-02
Silver
Palladium
Rhodium
* Heterogeneous
i< 0.01 ppm
> 100 ppm

k
1C*
TROGAAPMtC AMALYI
IN PPM WEIGHT
ELEMENT
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Approved: / >
/
77
MS
CONC

0.1

0.04
0.02

<0.01
0.3

0.02

0.02
0.7
0.5
1
0.04
10
0.2
0.4
VY //

°*e: September 22, 1980
Analyst: L. Jacobs
IAD No.: 97-E697-116-28
ELEMENT
Vanadium
Titanium
Scandium
Cal ci urn
Potassium
Chlorine
Sulfur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Hydrogen

CONC
0.02
5
<0.01
MC
7
*70
0.2
1
25
MC
2
4
=1
NR
NR
NR
0.04

0.02
NR


-------
00
       T^ Corporation
       ANALYSIS LABORATORIES
DATA REPORTING FORM
                      CUSTOMER
                                  CMEA
             nATF  July 10. 1980
CUS
RES
AC
TOMER COI
ULTS REPOI
lORPSS ....
MTRACT NO
UTTO
307605.22


ACUREX C(
TELEPHONE
)NTRACT N
n


Mountain View May 21, 1980 MfliO TU.«J/»tC \TWrEM

SAMPLE ID (CUSTOMER)
SAMPLE ID (LAB)
PARAMETER
Blank
Sample Aliquot
Total Sample
Total Sample
Total Sample








Fuel Hg
-!W--.-.-.mrr::--:r:.--i-----!:--?--
FMF_ — -i— ~
Less 1
Less 1

Less 0.1









XAD Hg
Less 1
Less 1

Less 0.1
Less 10








1 Imp Hg
Less 1
Less 1
Less 1

Less 1








2&3 Hg

Less 1
Less 1
Less 1

Less 1








2&3 As
' i " . L • " _ •
;:;T:;-V— .«:: • —_-.._ —
Less 10
Less 10
Less 10

Less 10








2&3 Sb

..'- -_^_^'_ • '• '
!-r'-g-L-;v"E" 	 '
...^-M-.— .... - -.
Less 3
Less 3
Less 3

Less 3








*A11 water tank samples were below 1 ug/L
form EED-06T 4/60
Water Hg

=5==ss=EE±=E==
Less 1*
Less 1
Less 1











^=
3




























f^^^^^^mmummfmtmnflff
UNITS
ug/L
ug/L
ug/L
ug/g
ug








ANAIVST (r- fa* 4 (/
RFMIFWPR ^£*U& ~>t*^€f

-------
            COMMERCIAL TESTING  & ENGINEERING CO.
            OtMtlUl OfHCtS m MOXTM U JAUt ITMtT, CM.CAaO. ILUHOU M«ei  .  ME* COOlT " ^
Reply to
Instrume
490 Orch
Golden. CO 80401
Imtrumentil An»Jytis Division                                                        m^. ao3.27B „-„
490 Orchard Street                                                                ^)ne 303-278-9521
                                     November  18.  1980
             Mr.  Brent Higginbotham
             Acurex Aerotherm Corp.
             485  Clyde Avenue
             Mountain View, CA  94042

             RE:   IAD #97-E697-116-28
                                      Analytical Report

            We  were asked as per phone conversation with  Carlo  Castaldini,  on
            October 20, 1980, to check by atomic absorption, several elemental
            values  on samples that were analyzed by spark source mass spectrometry
            (SSMS)  and reported September 24, 1980.

            Nickel  was checked on the XAD Resins Acurex ID Nos. 80-1473 & 1533-37.
            Nickel  contamination was due to combustion in the Parr combustion
            apparatus as they were prepared for SSMS.  The samples were reprepped
            by  ashing at approximately 600° C, dissolving the ash in aqua regia,
            and diluting with deionized water.  Nickel was determined from  this
            solution as well by flame atomic absorption.

            Copper  was found to be a major component on samples 80-1368 and 80-1369
            by  SSMS.  This was quantitated by flame atomic absorption on a  diluted
            portion of each of the samples.

            The results of these analyses are presented in Table No. 1 and  are re-
            ported  in the appropriate units on the samples "as received".
                                            I. OH . Olimo. CB > OOLMN. CO • Mm«ON. IT . JU>». >t •
                                            . MUVN.ll.«» 'MUM. UT •». NOIUU*. IL.TOtteO.O". MHOMim. l£. CM.
                                           79

-------
                                            Table No. 1
                                            Nickel
                 Sample »                   lug/9)
                 80-1473 fW«  **>           2.4
                 80-1368 To~*fc»*»* Prior     4* —                      505
                 80-1369 Tcut-oA^af^-U^- —                      480


                 If there are any questions concerning these results, please call
                 Bruce A. Hale                             M. L. Jacaps. Ph.D., l»Ingr.
                 Section Supervisor                        Instrumental Analysis Div.
                 as
                            COMMERCIAL TMTINO ft •NaiNEIKINO CO.
Onginil Copy Wittrmtrlod
  for You' Prottclion

Hit
                                                  80

-------
                                       1C  ANALYSIS  SHEET


          Acurex                         F.Cl.Br.NOg.NQ-,
Contractor                                                   	_^	


SimphSH. 	Mountain  View	 Simple Acquisition Oite    **  21* 198°
Typi of Source
TtstNumbtr - , — _ _ Simple ID Number _

.   , „   ...    Water Tank
Simple Description . - , _ _       _

.  ,  „     ...   E.  Hagmann              „   _         July  21,  1980       8pm - 2am
Anilyst Responsible - T - Dite Analyzed        _  Time _
                            G.  Nicoll                                        July 23,  1980
Calculations and Report Reviewed By .	  Report Dite	

            Micromeritics - Wescan Conductivity
Instrument	


PI....*  0.0025M  Sodium  Benzoate @ pH  6.0


               1.50 ml/mi n      _.„   1.08 KPSIp.M^g... *>* cm/min
Simple Size   IUW  "'	 Attenuator Setting	

                                                                                    X
Original Simple Volume or Mass 	 Multiple Stindird Addition: Yes	  No 	
 Observations
            30  min run, good separation
                                                 81

-------
                                        1C ANALYSIS SHEET
                                          so3,so4
Contactor  Acurex	
Sample s.a   Mountain  View	sample Ambition Oat.   ^ 21,  1980

Typt of Source -
T«$t Number	 Sample ID Number
                  Water Tank
Sample Description
Analyst Responsible  E' H«gnann	 D.t.An.ly»d   J"^   "' 198°  Tin,.


           Micromeritics -  Wescan  Conductivity
C.,cu«,tion,.ndR,portR8view«.By  G'  N1C011	  Report D.t.
Instrument.
         0.0047M KHP  @ pH 4.5
E luent	1	
               2.00 ml/min               0.79  KPSI                            1 cm/min
Column Flow Rate	 Pressure 	 Recorder Speed	
          10  Ul                                                 4
Sample Size  __	__ Attenuator Setting 	 	
Original Sample Volume or Mass 	    „   Multiple Standard Addition: Yes	  No
Observations      Spike  and recovery: average  recovery at 88%
                                                82

-------
                                        1C ANALYSIS SHEET

                                          PO,
Contactor	Acurex
*   ,.«..  Mountain View                     .   lm   ...  „      May 21,  1980
SainpltSitt 	_ Sample Acquisition Date      J    	
Type «f Source
Test Number	 Simple ID Number	


           .    Water Tank
Sample Description	,	    -     	


                  E. Hagmann                          July 22, 1980        8am  -  3pm
Analyst Responsible 	_	 Dite Analyzed           	 Time	
Calculations and Report Reviewed By  G.  NJCOll _ Report Date  July 23,  1980


instrument  Micromeri tics - Wescan  Conductivity _


        x 10"3M Phthalic Acid
                2.00ml/min               1-43 KPSI _  ,^,^   0.5 cm/mi n
 Sample Size         u' _  Attenuator Setting
 Original Sample Volume or Mass  	,	  Multiple Standard Addition: Yes	   No


            Spike and  recovery:  106% recovery
 Observations     r	±	

-------
           Water Tank  Blank-  Tap Water
Ion
F ~
d~
Br~
NOJ
NOJ
so3=
504
POf
Uncorrected
Simpli Value
ND
2
ND
ND
ND
ND
ND
ND
Blink
Value
ND
ND
ND
ND
ND
ND
ND
ND
Corrected
Sample Value

2






High/Low Calibration
Standards or Con-
ctntration Addad
5, 50
5, 50
5, 50
5, 50
5, 50
50
50, 500, 1000
10, 50
Dilution
Factor








Astignad
Concantration*
ND
2
\ ND
ND
ND
ND
ND
ND
Detection
Limit*
1
1
5
2
5
10
10
3
'Results: Hlg/L values (in original sample or I - Interference; MC - major constituent, not quantified; NC - not computed; NG - sample value below
         blank; ND - not detectable (<2 a blank or baseline).

-------
                                Water  Tank-  Sample Blank
oo
Ol
ton
F~
a"
Br~
M<£
N0§"
so3=
soj
»•
tin ••«••• ait
URCvrracmi
Simplt Value
ND
2
ND
ND
ND
ND
ND
ND
Blank
Vilut
ND
ND
ND
ND
ND
ND
ND
ND
Corrected
Simplt Vilui

2






High/Low Calibration
Standards or Con-
cantration Added
5, 50
5, 50
5, 50
5, 50
5, 50
50
50, 500, 1000
10, 50
Dilution
Factor








Alined
Concentration*
ND
2
ND
ND
ND
ND
ND
ND
Detection
Limit"
1
1
5
2
5
10
10
3
                      "Results:  mj/L values (in original sample or I - Interference; MC - major constituent, not quantified; NC - not computed; N6 - sample value below

                               blank; NO - not detectable (<2 a blank or baseline).

-------
                                Water  Tank -  Prior to Test
CO
en
Ion
F ~
d~
Br~
NOf
NOT,
sof
$04
POf
Itacomcted
Sample Value
ND
3
ND
ND
7
ND
990
ND
Blank
Value
ND
ND
ND
ND
ND
ND
ND
ND
Corrected
Sample Vilut

3


7

990

High/Low Calibration
Standard! or Con-
contrition Addad
5, 50
5, 50
5, 50
5, 50 .
5, 50
50
50, 500, 1000
10, 50
Dilution
Factor








Astignad
Concentration*
ND
3
'. ND
ND
7
ND
990
ND
Detection
Limit"
3
2
10
10
5
10
10
20
                      •Rtjulti: mj/L values (in original sample or I - Interference; MC - major constituent, not quantified; NC - not computed; N6 - sample value below

                              blank; ND - not detectable (<2 a blank or baseline).

-------
                                Water  Tank  - Sample
00
Ion
F~
a~
Br~
NOJ
N0£
so3=
804
POf
Uacornctid
Simph Value
ND
3
ND
ND
7
ND
1000
ND
Blank
Valin
ND
ND
ND
ND
ND
ND
ND
ND
Comcttd
Simple Vilui

3


7

\ooo ^

High/Low Calibrttion
Standards or Con-
centration Added
5, 50
5, 50
5, 50
5, 50
5, 50
50
50, 500, 1000
10, 50
Dilution
Factor








aaai-aaii
fuufMB
Concentration*
ND
ND
ND
ND
7
ND
1000
ND
Detection
Limit*
3
2
10
10
5
10
10
20
                      "Results: rtB/L values (in original sample or I - Interference; MC - major constituent, not quantified; NC - not computed; NG - sample value below
                               blank; NO - not detectable (<2 a blank or baseline).

-------
5.5    ORGANIC ANALYSIS REPORT
                                   88

-------
                    ORGANIC COMPOUNDS (bp <10(r C)
           rl^tAVtf v
    Ito    rfOu>»Tt«i'«  l/iViJ
             fy.wi   J
M«W*B Hri RtHK Ktrim^ IT
                         &
                                                        Tim
                                Workup
 1 MSohtaV)
                 1.0
 I EhanMOrlMlAtaVl

 7.

 &

It

U.
                K
                                    1 bikmiMDm

                                    1

                                     Ai

                                   11.
       mi irin (a wipnl atnph) »t I -
       •D - MI <«tKHMi «le M«k ir
                          iuafamm; HC - MI c*m»«4: NG - ami* rtut Mm Mmk;
                     Ttaw
                                •( Nik
                                        IK)
                                                                       CMC.
            K)0
                   0.7
0.087
                              Uo
                                             OM
   wo
                    11 fc.,-
                              MO
                                                         Ll_^_
                               K!D
                                                         U
            •00
  VJC
                                       89

-------
                      ORGANIC COMPOUNDS (bp Mnnil.M(aiUi.M
                   It O
 & EbcMaMBtalA/aV)

 7. kKphttntaU	

                                       4. CahMPmajnbnil-

                                       L MtnliaaDaM  —
   FkuMFInRanitail^aW:  Hj-
                               1 OHM TtMHiiUn fO

                                Air 	
11

11
                                      11.
                  >•»   t2.
                        / /*,O9D  -^
mi nte (» w<|iMl MiipM if I - tturttima: MC
•0 - Ml JmcttMi « J 0 Wnk ir Mint).
                                       - MI unf nN: KG - Miplt urin Mm Mnik;
                       ThM
                                            no
                                                              M|l/Laitealikntin
             AID
             0.7 HM
                            0.067
   UO
     Ml)
LO
                                                   an
                                           2-l.pp,
                      U  *,.;.
                         K)J)
                             0.17
H.X
                         m«
                                 .V/J)
                                           C.28
                                           l.i
   WD
                         Aid
                             o.n
b
                                 MO
                                                        III
   U6
                                                                    ^-
                                            90

-------
                     ORGANIC COMPOUNDS (bp <100* C)
TypirflMfw


tatlkmtw
                                                                   AV/g
                                                                  o
-------
           ACUREX.
           Corporation
       ANALYSIS LABORATORIES
                        CUSTOMER
CNEA
 DATA REPORTING FORM

	  DATE   July 15. 1980
                        CUSTOMER CONTRACT NO.  307605.22
                        RESULTS REPORT TO 	
                          ADDRESS 	
                          ACUREX CONTRACT NO.
                          TELEPHONE 	
                            Mountain View May 21. 1980
SAMPLE 10 (CUSTOMER)
SAMPLE ID (LAB)
TCO Blank
TCO Sample
TCO, Corrected
GRAV Blank
GRAV Sample
GRAV, Corrected
TCO & GRAV
TCO & GRAV
Volume Analyzed




Filter

	 	


Less 2
3
3
3
0.1





XAD

0.07
74
74
4
22
18
92
3.2





OMC

Less 0.02
Less 0.02
Less 0.02
Less 2
5
5
5
0.2





Tank
Blank

Less 0.02
Less 0.02
Less 0.02
Less 2
6
6
6
0.2
3665




Tap
Water

Less 0.02
Less 0.02
Less 0.02
Less 2
Less 2
Less 2
Less 2
Less 0.1
3720




Prior to
Test

— r~ r — • -a— rr:
Less 0.02
Less 0.02
Less 0.02
Less 2
Less 2
Less 2
Less 2
Less 0.1
1300




Tank
Sample

Less 0.02
0.50
0.50
Less 2
Less 2
Less 2
Less 2
Less 0.1
10,000





















. — ...i-i. . TTTT •-;;














mg
mg
mg
mg
mg
mg
mg
mg/dscm
ml




vo
       FormEEO-097 t to
                                                                               ANALYST .

                                                                               REVIEWER
                                                                                      M. Gardner, G.  Nlcoll

-------
Ill REPORT
             Filter Mountain View 5/21/80
                                    Ho Peaks
                                        93

-------
Ill ftC'OUT
            Filter Blank
   We* R«>tat
hmwiiy
                                    No Peaks
                                         94

-------
IM MPOftT
SAMPLE:	
XU> Mountain View 5/21/80
!W
3575-3070
2905
2845
2750-2450
1725
1445.
1265
1095
805























tafMitr
w
s
s
M
M
w
w
N
W























Anipment ComnMBti
O-H
C-B Alkane
C-H Alkane
O-H
OO
C-H Alkane
C-0 .
C-0
Not Assigned

•


•• •


















                                        95

-------
IAMPLI:     XAD BlanX
                    totraity
                                  No Peaks
                                        96

-------
in REPORT
            OMC Mountain View 5/21/80
                    tamwity
                                    Mo Peaks
                                         97

-------
1AMELI:     Tap Water Mountain Vi«* 5/21/80
                                   No Peaks
                                                                  CemmMi*
                                          98

-------
Ill REPORT
             Tank Blank  Mountain View 4/21/80
      «-•'»
                     brevity
                                     Mo Peaks
                                          99

-------
Tank Sample  Mountain Vi«* 5/2X/80

!~j*r
2905
2845
1190


.


























•_*,
S
S
S







•





















M&*^tfMMt BWWWfli
C-B Alkane
C-B Alkane
Sulfonamide Found in GC/MS PHA







.


•• •








•


•


•
.


                           100

-------
                    COMPOUNDS SOUGHT
                    Detection Limit  in nanograms
 8
 1
 2
 8
40
 1
 1
 8
 4
40
  3
  3
  1
  2
  2
  2
  5
  7
  8
  8
  5
  1
  1
 40
  2
  1
 40
* 4-bromophenyl phenyl ether
* bis (2-chloroisopropyl) ether
* bis (2-chloroethoxy) methane
* hexachlorobutadiene
* hexachlorocyclopentadiene
* isophorone
* naphthalene
* nitrobenzene
* N-nitrosodiphenylamine
* N-nitrosodi-n-propylaroine
* bis (2-ethylhexyl)  phthalate
* butyl benzyl  phthalate
* di-n-bujyl  phthalate
* di-n-octyl phthalate
* diethyl  phthalate
* dimethyl phthalate
* benzo(a)anthracene
* benzo(a)pyrene
* 3,4-benzofluoranthene
* benzo(k)fluoranthene
* chrysene
*  acenaphthylene
*  anthracene
* benzo(ghi)perylene
* fluorene
 *  phenanthrene
 *  dibenzo(a,h)anthracene
 40
  2
 20
  2
100
  8
  8
  8
  3
  2
  4
  8
  4
 20
 10
 10
   1
   2
   4
 40
 40
   9
   @
  40
  40
  40
  40
* 1ndeno(1,2,3-cd)pyrene
* pyrene
2,3,7,8-tetrachlorodibenzo-p-dioxin
* acenaphthene
* benzidine
* 1,2,4-trichlorobenzene
* hexachlorobenzene
* hexachloroethane
* bis(2-chloroethyl)ether
* 2-chlpronaphthalene
* 1,2-dichlorobenzene
* 1,3-dichlorobenzene
* 1,4-dichlorobenzene
* 3,3-dichlorobenzindine
* 2,4-dinitrotoluene
* 2,6-dinitrotoluene
* 1,2-diphenylhydrazine (as azobenzene)
* fluoranthene
* 4-chlorophenyl  phenyl ether
anthanthrene
benzo(e)pyrene
dibenzo(a.H)pyrene
dibenzo(a,i)pyrene
dibenzo(c,g}carbozole
7,12  dimethyl  benz(a)anthracene
3-methyl  cholanthrene
perylene
 *Authenic standard ran
 Wtolecular weight too high for direct
                                         40  Benzo(c)phenanthrene
                                  analysis by Base/Neutral  run
                                        101

-------
        ^Corporation
        ANALYSIS LABORATORIES
                                     CMEA
                         CUSTOMER
                         CUSTOMER CONTRACT NO.   307605.22
                         RESULTS REPORT TO 	
DATA REPORTING FORM
                      July 21. 1980
               DATE
               ACUREX CONTRACT NO.
               TELEPHONE 	
                           ADDRESS
                              Mountain View 5/21/80
        28.5 dscm
1 SAMPLE ID (CUSTOMER)
SAMPLE 10 (LAB)
PARAMETER
Amount Infected
Naphthalene*
Phenanthrene/
Anthracene*
Naphthalene
Phenanthrene/
Anthracene
Volume analyzed





Filter Bk

	 	 — — -7T- •
"'"'L,.1™1.' ."'._•::.::
^^
2
Less 0.5

Less 0.5









Filter


2
Less 0.5

Less 0.5
Less 0.02

Less 0.02






XAD Blank

••sr7sr:B-,;^T-!BT-.-;.'.-r-r:
2
Less 0.5

Less 0.5









Xad

-'-"'-':: '--:'•• •"•-":"'' ' WS""-jr ;
1
94

4
36

2






OMC


2
Less 0.5

Less 0.5
Less 0.0;

Less 0.0;






9
Mater Blk


- -'--„-- v- i 'vi j -a .""— " •
3
Less 0.3

Less 0.3









Tank watei


.1
9

2
0.4

0.08
10,000























• '"• '^i~" i .. ' . ....
L 	 _g














UNITS
II 	 •l-IIM
uL
mg/L

mg/L
ug/dscm

ug/dscm
mL





o
ro
         *Sample Aliquot   0A11 water tank blanks are clean
        FwmEED-057 4/M
                                                                                 ANALYST _

                                                                                 REVIEWER .

-------
               SAMPLE:     Man   XAD  11473
o
CO

Mta*'
T*MtorLC*
i
TCO
•1
74
36
26
BMV
"•1
18
9
14
TCO+GIIAV
Tcttlnf
92
45
40
mg/dscm
3.2
1.6
1.4
wVHRMw
1 .
i
i
4
I
•
f
SMI
TtfthMj
FfKtMft
25
0.09
0.41
0.06
<0.01
<0.01
<0.01
26
MM*
<0.02
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.08
tor-
fMAM
25
0.09
0.41
0.06
cO.Ol
eO. 01
cO.01
26
W
52
0.2
' 0.85
0.1
<0.02
<0.02

-------
II
I


































1 REPORT
AMPLE: '

WvM MMMF
'«-•')
2920
2840
'


.









.




.-
. *










IAN XAD 1473 F1

knmftr
*
s
s

.






, •








t




... . _
•






"
.•
• Mf^_^_u^ ^^MMIkMlM
jWB^ViiiMii* w^^nnwniB
CH A3 iphatic hydrocarbons
CH Aliphatic hydrocarbons

•

-
r
•


.-


•• •









-"
	 .. ._ * - ._ 	 	 	
.
•

•



104

-------
•ft REPORT
                  MM  XAD  1473  F2
                                  No Peaks
                                      105

-------
Ill KPQftT
                                Mo Peak*
                                   106

-------
.     HAN XAD 1473 F4
                          No Peaks
                                   107

-------
MftEPOKT
•AMPU;	
HAN  XAD  1473  F5
                               tto Peaks
                                        108

-------
IftKEPONT
      .   MAN  XAD  1473  F6
                                o  Peaks
                                      109

-------
tmttro*T
SAMPLE:	
WN  XAD 1473  F7
                             No Peaks
                                   no

-------
LftMS REPORT
SAMPLE:	
                      1473 Fl Mountain View XAD
tenmifv
1





e»iw»
Aliphatic hydrocarbons

.

.

MWR.V






 Sub-Ciwgofiw. Specific Campoundt
    Ifitsfwty
                             Ctngery
m/i       ComDotition
Oth«r
                                           111

-------
5.6    BIOASSAY ANALYSES REPORT
                                     112

-------
                         GENETICS ASSAY  NO.   5397
                                            WiMMMMBHB

                              LBI SAFETY  NO.   6391
      CYTOTOXIC EVALUATION OF

           SAMPLE #1473
       (CMEA MOUNTAIN VIEW)
              IN THE
         RODENT CELL (CHO)
       CLONAL TOXICITY ASSAY
           FINAL REPORT
          SUBMITTED TO:

       ACUREX CORPORATION
        485 CLYDE AVENUE
MOUNTAIN VIEW, CALIFORNIA 94042
          SUBMITTED BY:

      LITTON BIONETICS,  INC.
        5516 NICHOLSON LANE
    KENSINGTON,  MARYLAND  20795

       LBI PROJECT NO.  20933

   REPORT  DATE:   DECEMBER, 1980

             113

-------
                               PREFACE

This report contains a summary of the data compiled during the evaluation
of the test compound.   The report is organized to present the results in
a concise and easily interpretable manner.  The first part contains items
I - IX.  Items I - IV provide sponsor and compound identification infor-
mation, type of assay, and the assay design reference number.  All assay
design references indicate a standard procedure described in the Litton
Bionetics, Inc. "Screening Program for the Identification of Potential
Mutagens and Carcinogens."  Item V provides the initiation and completion
dates for the study, and Item VI provides identification of supervisory
personnel.  Item VII identifies the tables and figures containing the data
used by the study director in interpreting the test results.   The interpre-
tation itself is in Item VIII.  Item IX provides the conclusion and evalua-
tion.

The second part of the report, entitled Assay Design, describes the
materials and procedures employed in conducting the assay.   This part
of the report also contains any appendices, as well as evaluation
criteria used by the study director.   The evaluation criteria are
included to acquaint the sponsor with the methods used to develop and
analyze the test results.

All test and control results presented in this report are supported by
fully documented raw data which are permanently maintained in the files
of the Department of Genetics and Cell Biology or in the archives of
Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington, Maryland, 20795.

Copies of raw data will be supplied to the sponsor upon request.
                                 114

-------
   I.         SPONSOR:    Acurex  Corporation

  II.         MATERIAL (TEST COMPOUND):  GENETICS ASSAY NUMBER:  5397

             A.    Identification:   Sample #1473 (CMEA Mountain View)

             B.    Date Received:    October 9,  1980

             C.    Physical Description:  Brown, opaque liquid

 III.         TYPE OF ASSAY:  Rodent Cell (CHO)  Clonal  Toxicity Assay

  IV.         ASSAY DESIGN NUMBER:  442

   V.         STUDY DATES:

             A.    Initiation:  October 20, 1980

             B.    Completion:  November 24, 1980

  VI.         SUPERVISORY PERSONNEL:

             A.    Study Director:  Brian C. Myhr, Ph.D.

             B.    Laboratory Supervisor:  Robert Young, M.S.

 VII.         RESULTS:

             The data are presented in Table 1 on page  4  and  in Figure 1
             on page 5.

VIII.         INTERPRETATION OF RESULTS:

             The test material, Sample No. 1473, was supplied as  a SASS
             train organic extract in 10 ml of methylene chloride.  The
             total organic content was given as 26 mg.  A portion of the
             sample (8.20 ml) was solvent exchanged into an equal volume
             of dimethylsulfoxide (DMSO).  The concentration of organics
             was therefore unchanged, and the value of 2.6 mg/ml  (or
             2.6 yg/vl) was used to convert the doses obtained  on a
             volume basis into the equivalent amount of organics  per
             milliliter of culture medium.

             The test material remained completely soluble after exchange
             into DMSO.  Diluted stocks were prepared with DMSO just prior
             to testing, and the treatments were initiated with culture
             media containing 1:100 dilutions of the stocks.  The highest
             test concentration was achieved with a 1:50 dilution of the
             solvent-exchanged sample, which introduced 2% DMSO into the
             medium.  Vetiicle control cultures were therefore exposed to
                                     115

-------
VIII.  INTERPRETATION OF RESULTS:   (continued)

       culture media containing 1% and 2% DMSO  in order to provide the
       reference points for determining the survival  of the cells to
       treatments with the test material.  No precipitation of test
       material in the culture medium was observed at any of the tested
       concentrations.

       Seven test concentrations from 0.05 ul/ml  to 20 pi/ml  were
       evaluated for their effects on colony survival.   As shown in
       Table 1  and Figure 1,  none  of the assayed  concentrations  caused
       significant changes in the  numbers of colonies.   This  lack of
       observable toxicity was not conclusive,  however,  because  the
       highest concentration  corresponded to only 52  ^g  of organics/ml,
       which was essentially  the midpoint of the  concentration range
       where an EC50 would yield a moderate toxicity  classification (see
       Evaluation Criteria).   A sharp increase  in toxicity could occur
       with a small  increase  in concentration,  but it is reasonable to
       assume that 50% survival  would not have  been observed  had the high
       dose been doubled.   This concentration would have fallen  just
       within the low toxicity region.   Thus, the test material  was
       evaluated as  possibly  having low toxicity  or no detectable toxicity
       to CHO cells.   The position of the EC50  could  be  further  investi-
       gated by concentrating the  extracted organic material.
                                   116

-------
IX.  CONCLUSIONS:

     The test material, Sample No. 1473, was evaluated as having low or
     nondetectable toxicity in the CHO Clonal Toxicity Assay.   No
     toxicity was observed for concentrations up to the highest testable
     level of 52 yg of organics/ml.
                                      Submitted by:

                                      Study Director
Brian C. Myhr,'Ptf.D7
Section Chief
Mammalian Genetics
Department of Genetics
 and Cell Biology
                                       Reviewed  by:
                                                                U/i/Vo
                                                                  Date
                                       David  J.  Brusick, Ph.D.
                                       Di rector
                                       Department  of  Genetics
                                        and Cell Biology
                                   117

-------
                                     TABLE  1

                     RODENT CELL (CHO) CLONAL TOXICITY ASSAY
Sample Ide
MOUNTAIN V
ntity: Samole
'IEW
Description of Test Sam
yellow solution In DMSO
LBI Assay
No.: 5397
f!473. CMEA


iple: Clear, Dale
i.



Date Received: October 9, 1980
Test Date:
Vehicle:
Cell Type:
November 17, 1980
DMSO
CHO-K1
Cells Seeded per Dish:


200



EC50
Value: >2(
Toxicity
Classification
3 yl/ml (>5I
2 yg/ml)
: Probablv Low or Non-
detectable
pH Alterations
Comments on Tr<








: NONE
jatment:









'



CLONAL TOXICITY DATA
Sampl e
NC
VC, 1%
VC, 2%
Test
Test
Test
Test
Test
Test
Test
Applied
Concentration
pi /ml
10.0
20.0
0.05
0.2
0.5
2.0
5.0
10.0
20.0
Dish
#1
207
180
173
183
C
185
161
161
166
.170
Dish
#2
185
177
162
192
192
189
187
190
178
171
Dish
#3
192
172
166
189
183
179
183
164
180
154
Average
Count
194.7
176.3
167.0
188.0
187.5
184.3
177.0
171.7
174.7
165.0
Relative
Survival*
%
100.0
100.0
106.6
106.4
104.5
100.4
97.4
99.1
98.8
Cloning
Efficiency

-------
                                         FIGURE  1


                            RODENT CELL  (CHO) CLONAL TOXICITY ASSAY

                                     EC50 DETERMINATION


                         SAMPLE  #1473,  CMEA  MOUNTAIN VIEW
   140


   130


   120


   110


   100


    90
»«   80
 •


§   70
be
i
60


50


40


30


20


10
      0.1
                                                      10
100
                                     CONCENTRATION,   pi/ml
                                         119

-------
                             ASSAY DESIGN NO. 442

                    RODENT CELL CLONAL (CHO) TOXICITY ASSAY


This assay conforms to one of the bioassay tests in the US Environmental Protec-
tion Agency (EPA) IERL - RTP Procedures Manual:   Level 1 Environmental Assessment
Biological Tests (September, 1980).Level 1 bioassays obtain preliminary infor-
mation on the harmful biological effects of chemicals found in industrial feed
and waste streams by using mammalian cells in culture to measure metabolic
impairment and cellular death.


1.   OBJECTIVE

     The objective of this assay is  to determine the concentration of a test
     article that will reduce the colony forming ability of Chinese hamster
     cells by 50% after a 24-hour exposure.   This concentration is referred to
     as the EC50 value.


2.   MATERIALS

     A.   Indicator Cells

          The indicator cells used in this study are Chinese hamster CHO-K1
          cells (American Type  Culture Collection No.  CCL 6l).   This cell type
          was derived from ovarian tissue and has spontaneously transformed to
          a stable,  hypodiploid line of rounded,  fibroblastic cells with un-
          limited growth potential.   Monolayer cultures  have a fast doubling
          time of 11 to 14 hours,  and untreated  cells can normally be cloned
          with an efficiency of 80%  or greater.   Permanent stocks  are main-
          tained in  liquid nitrogen  and laboratory cultures are maintained by
          serial  subculturing.   Laboratory cultures are  periodically checked
          by culturing methods  for the absence of mycoplasma contamination.
          This test  system is specified by the IERL- RTP Procedures Manual.

     B.    Medium

          The  CHO-K1 cell  line  has an absolute requirement for proline and
          therefore  must.be  maintained in a  culture medium containing suf-
          ficient amounts  of this  amino acid.  Ham's F12 medium, which contains
          3 x  10-«M  L-proline,  is  used,  supplemented with 10% fetal bovine
          serum,  100 units per  milliliter of penicillin,  100 ug streptomycin
          per  milliliter,  and 0.5  ug  amphotericin B (Fungizone) per milliliter.

    C.    Controls

          Untreated  cells  are cloned  to establish the control  cloning effi-
          ciency.  If the test  article  is  dissolved in an organic  solvent
          (usually dimethylsulfoxide),  cells  exposed to  solvent in the medium
                                      120

-------
2.   MATERIALS (Continued)

          are cloned to provide the reference cloning efficiency for the effect
          of the test article.  The final concentration of solvent in the growth
          nedium is generally IX or less.  All controls are performed in tripli-
          C A v6 •

     D.   Sample Forms

          Solid samples are tested as a solution or are ground to fine particles
          (less than 5 urn) and tested as a suspension in growth medium.  Dry
          particulate articles, aqueous liquids, suspensions and slurries are
          added directly to the growth medium and tested as a suspension or
          solution.  Liquids containing less than 0.2% organic solvent are
          generally tested directly; samples dissolved in organic solvents are
          solvent exchanged in dimethylsulfoxide (DMSO) before testing.  Original
          sample volumes may be reduced a maximum of 10-fold during solvent
          exchange, and the concentration factor is used to convert assayed
          volumes into equivalent original sample volumes.  All sample manipu-
          lations are performed as described in the IERL-RTP Level 1 Procedures
          Manual.


3.  'EXPERIMENTAL DESIGN

     A.   Dose Selection

          Unless the approximate toxicity is already known or the sample size
          is limiting, the following dose ranges are tested for different sample
          forms.  Dry particulate articles are dissolved or suspended in growth
          medium and tested at five dose levels from 1000 ug/ml to 10 pg/ml.
          Aqueous samples, suspensions or slurries are tested from 600 Hliters/ml
          to 6 |jliters/ml in five dose steps.  Samples that are solvent exchanged
          into DMSO are tested from 20 uliters/ml to 0.2 uliters/ml, also in five
          dose steps.  Solvent concentrations are IX for all dose levels except
          the maximum applicable dose (MAD) of 20 ul/ml, which contains 2% solvent.
          A second dose study is performed with an appropriate dose range if the
          EC50 has not been properly located in the initial test; EC50 values
          greater than 1,000 ug/ml, 600 |jliters of aqueous sample/ml or 20 uliters
          nonaqueous sample/ml are not determined.

     B.   Clonal Toxicity Assay

          Cells from monolayer stock cultures in logarithmic growth phase are
          trypsinized, counted by hemacytometer and reseeded into a series of
          60- or 100-mm culture dishes at 200 cells per dish.  The cultures
          are incubated for 6 to 16 hours at 37°C to allow attachment of the
          cells and recovery of growth rate.

          Test article is then applied (three dishes per dose), and the cultures
          returned to the incubator.  If the test article causes a color change,
          an additional dish is treated with the high dose and the pH of the


                                       121

-------
3.    EXPERIMENTAL DESIGN (Continued)

          medium 1s determined.   The  pH is also recorded for the highest dose
          which results in a slight color change.   After a 24-hour exposure
          period the medium is aspirated and the cells washed with Dulbecco s
          phosphate buffered saline (PBS; prewarmed to 37°C).   The pH of the
          discarded medium for which  initial pH measurements were made is again
          recorded.  Fresh medium is  placed on each culture and Incubation con-
          tinued for an additional  6  days to allow colony development.  Medium
          is drained from the cultures and the surviving colonies are washed
          with PBS, fixed in methanol, and stained with Giemsa.  Colony counting
          is performed by eye; colonies smaller than 50 to 100 cells are not
          counted.


4.   ASSAY ACCEPTANCE CRITERIA

     The assay is considered acceptable for evaluation of the test results if
     the following criteria are met:

               The average cloning efficiency of the CHO-K1 cells in the nega-
               tive controls is 70% or greater, but not exceeding 115%.

               The distribution of colonies in the treated cultures is generally
               uniform over the surface of the culture dish.

               The data points for each test concentration critical to the loca-
               tion of the ECSO are .the averages of at least two treated cultures.

               A sufficient number of test concentrations are available to
               clearly locate the EC50 within a toxicity region as defined
               under Assay Evaluation Criteria.

               If the EC50 value is greater than 1000 ug/ml, 600 uliters Of
               aqueous sample/ml, or 20 uliters of nonaqueous  sample/ml, the
               plotted curve does not exceed 110% of the negative control.


 5.   ASSAY  EVALUATION CRITERIA

     The  screened doses,  pH values (if appropriate), colony counts,  percent
     survivals (colony counts relative to control colony counts)  and ECSO
     values are  provided.  The percent survival is plotted as  a function of
     applied concentration and the EC50  value  determined graphically by fitting
     a curve by  eye through the  data points.   The  EC50  1s  used to rank the test
     material  using the  standard evaluation criteria  defined  in the table below.

     Sorbent extracts  of known organic content are evaluated  as nonaqueous
     liquids and In terms of  their solid contents  (ug organics/ml), and are
     ranked using the  more  sensitive  parameter.  Where data on the original
     liquid or gas sample volumes  is  provided  by the  Sponsor, the EC50 value
     is calculated in  terms of these  equivalent volumes per mill inter of
     culture medium; however,  no evaluations  are made as yet on this basis.


                                       122

-------
5.    ASSAY EVALUATION CRITERIA (Continued)
        Toxicity*
     Solids
(EC50 in ug/ml)
Aqueous Liquids
(EC50 in
Nonaqueous Liquids
 (EC50 in ul/ml)
High
Moderate
Low
Not Detectable
10 to 100
100 to 1000
>1000
<6
6 to 60
60 to 600
>600
.2-2
2-20
>20

     Evaluation criteria  formulated  by  Litton Bionetics, Inc. for IERL-RTP
      Procedures Manual:   Level  1  Environmental Assessment Biological Tests.

      Criteria for  nonaqueous  liquids are  tentative and under evaluation.
     RECORDS TO BE MAINTAINED

     All raw data, protocols,  protocol  modifications, test article weight and
     dispensation records  and  correspondence  between  LBI and the Sponsor are
     being maintained  in a central  file within  the Department of Genetics and
     Cell Biology.  These  records will  be  filed under departmental assay number
     and held  up to 2  years following submission of the final report to the
     Sponsor.  After 2 years they will  be  transferred to the LBI archives for
     permanent storage.
                                        123

-------
                      Q.A.  Inspection  Statement
                   (reference 21  CFR 58.35(b)(7))
PROJECT    J0993    '"                      LBI Assay No.
TYPE of STUDY  fydi^lJr Csjfi fat))  C£jnu>JL.
         This final  study report was reviewed by the LBI Quality
Assurance Unit on /UatnukeA^ I iQfo.  A report of findings was
                                 ;
submitted to the Study Director and to Management on
         The short-term nature' of this study precluded inspection while
it was in process.   The Quality Assurance Unit inspects an in-process
study of this type  approximately once per month to assure that no
significant problems exist that are likely to affect the integrity of
this type of study.
                                     iuubu,*ju
                                     Auditor, Quality Assurance  Unit
                                   124

-------
                        GENETICS ASSAY NO.    5397
                             LBI SAFETY NO.   6391
    MUTA6ENICITY EVALUATION OF
           SAMPLE #1473
              IN THE
    AMES SALMORELOTHICROSOME
         "~PlATE"TEST
          FINAL REPORT
          SUBMITTED TO:
       ACUREX CORPORATION
        485 CLYDE AVENUE
MOUNTAIN VIEW, CALIFORNIA   94042
          SUBMITTED BY:
     LITTON BIONETICS,  INC.
      5516 NICHOLSON LANE
    KENSINGTON, MARYLAND  20795
      LBI PROJECT NO. 20988
      REPORT DATE:  DECEMBER,  1980

               125

-------
                        PREFACE


This report contains a summary of the data compiled during the
evaluation of the test compound.   The report is organized to
present the results in a concise  and easily interpretable manner.
The first part contains items I-IX.   Items I-IV provide sponsor
and compound identification information, type of assay, and the
protocol reference number.  All protocol references indicate a
standard procedure described in the Litton Bionetics, Inc.
"Screening Program for the Identification of Potential Mutagens
and Carcinogens."  Item V provides the initiation and completion
dates for the study, and Item VI  provides identification of super-
visory personnel.  Item VII identifies the tables and/or figures
containing the data used by the study director in interpreting
the test results.  The interpretation itself is in Item VIII.
Item IX provides the conclusion and evaluation.

The second part of the report describes the materials and procedures
employed in conducting the assay.  This part of the report also
contains evaluation criteria used by the study director, and any
appendices.  The evaluation criteria are included to acquaint the
sponsor with the methods used to develop and analyze the test
results.

All test and control results presented in this report are supported
by fully documented raw data which are permanently maintained in
the files of the Department of Genetics and Cell Biology or in the
archives of Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington,
Maryland  20795.
                              126

-------
   I.   SPONSOR:   Acurex Corporation

  II.   MATERIAL  (TEST COMPOUND):  GENETICS ASSAY NUMBER:   5397

       A.   Identification:  Sample #1473

       B.   Date  Received:  October 9, 1980

       C.   Physical  Description:  Pale yellow liquid

 III.   TYPE OF ASSAY:  Ames Salmonella/microsome Mutagenesis Assay

  IV.   PROTOCOL  NUMBER:  401 (EPA-Level I)

   V.   STUDY DATES:

       A.   Initiation:  November 7, 1980

       B.   Completion:  November 13, 1980

  VI.   STUDY DIRECTOR:  D.R. Jagannath, Ph.D.

 VII.   RESULTS:

       The results of this assay are presented in Table 1.

VIII.   INTERPRETATION OF RESULTS:

       The test  material, Sample No. 1473, was supplied as a SASS train
       organic extract in the 10 ml of methylene chloride.  The total
       organic content was given as 26.0 mg.   A portion of the sample
       (8.20 ml) was solvent exchanged into dimethylsulfoxide (DMSO) to
       a final volume of 8.20 ml.  This solvent exchanged sample (test
       material) was examined for mutagenic activity  in the Ames/Salmonella
       assay in  the  presence and absence of liver microsomal enzyme pre-
       parations from Aroclor-induced rats.

       A negative control consisting of the solvent DMSO and-specific
       positive  compounds were also assayed concurrently with the test
       material.

       DOSE RANGE:

       The dose  range employed for the evaluation of  this test material
       was from  10.0 yl to 200.0 yl per plate.  Based on the organic
       content of the sample the doses employed would equal to 26.0 vg to
       520.0 yg  per  plate.  The  tests were conducted  using two plates per
       dose level.
                                     127

-------
VIII.  INTERPRETATION OF RESULTS:   (continued)

       TOXICITY

       The test material did not exhibit  toxicity with  any of the
       indicator strains used in this  assay.

       The results of the tests conducted on the test material  in the
       absence of an activation system was positive with  the  strain  TA-98.
       The lowest dose at which the response was observed was at  100 yl
       or 260.0 yg organics  per plate.

       The results of the test conducted  on the test material  in  the
       presence of rat liver activation system were negative.

  IX.  CONCLUSIONS:

       (1)  The SASS train organic  extract sample #1473 exhibited  genetic
       activity with the strain TA-98  in  the nonactivation  assays  conducted
       in this evaluation and is considered as mutagenic  under  these test
       conditions.   (2)   Based on the  EPA's 'Definition of  Toxicity
       Categories for Health Effects Tests' the mutagenic activity of the
       sample is classified  as 'Moderate1 (see Table A).  The specific Acti-
       vity at the Minimum Effective Concentration (which is  the  number of >e-
       vertants minus the background revertants divided by  yg organics x 1000) of
       the sample (260.0 yg) based  on  the results from TA-98  (nonactivation)
       was 92.31  revertants  per mg  of  organics.

       The classification of the mutagenic activity (item 2 above) and the
       Specific Activity of  the sample (item 3 above) are given here to
       compare the potency of various  samples belonging to  the  same  class.

                                 Submitted  by:

                                 Study Director
                                 D.R.  Jagannath,  Ph.D.     Date
                                 Section  Chief
                                 Submammalian  Genetics
                                 Department  of Genetics
                                 and Cell  Biology

                                 Reviewed  by:
                                                  f^f f     - - -  i i^
                                                _  Ph.D.  Tate
                                Director
                                Department of Genetics
                                and Cell Biology
                           128

-------
                                                                   TABLE A
                                           DEFINITION OF TOXICITY CATEGORIES FOR HEALTH EFFECTS ASSAYS*
PO
Assay"
Ames
RAN
CHO
WAT
"standard
Ames:
Activity Measured6
NEC
(mutagenesls)
(lethality)
EC50
(lethality)
LD80
(lethality and
toxic signs)
test abbreviations are as
Ames Salmonella/microsome
Sample
Typec
S
AL.NAL
S
AL
NAL
S
AL
NAL
S
AL.NAL
follows:
mutagenesls
MADd
5
200
1
600
20
1
600
20
5
5
assay
Range of Concentration or
Units
mg/plate
Ml/plate
Ml/ml
Ml/ml
mg/ml
Ml /ml
Ml /ml
gm/kg
ml/kg

High
<0.05
<2
<0.01
<6
<0.2
<0.01
<6
<0.2
<0.05
<0.05

Node rate
0.05-0.5
2-20
0.01-0.1
6-60
0.2-2
0.01-0.1
6-60
0.2-2
0.05-0.5
0.5-0.5

Low
0.5-5
20-200
0.1-1
60-600
2-20
0.1-1
60-600
2-20
0.5-5
0.5-5

Dosage
Not Detectable
(ND)
ND at >5
ND at >200
ND at >1
ND at >600
ND at >20
ND at >1
ND at <600
NO at >20
ND at <5
ND at <5

                  RAN:  Rabbit alveolar macrophage cytotoxicity assay
                  CHO:  Rodent cell clonal toxlcity assay
                  WAT:  Acute |n vivo test In rodents (whole animal test)

              Standard abbreviations for measured endpoints are as follows:
                  NEC:  Minimum effective concentration
                 EC90:  Calculated concentration expected to produce effect 1n 50 percent of population
                 LDSO:  Calculated dose expected to kill 50 percent of population

             CS = Solid, AL - Aqueous liquid, NAL = Nonaqueous liquid

             
-------
          RESULTS
                                                         TABLE I
              NAME OR CODE DESIONATION OP THE TEST COMPOUND:  SIMPLE f!4W
              SOLVENT:  OHSO
              TEST INITIATION DATES:  tl/OT/HO
              TEST COUPLET ION DATE: 11/13/RO
              s-9 LOTR: 00009
       NOTE:   CONCENTRATIONS *NE OIVEN IN NICROLITERS   PER  PLATE
       TEST
       NONACTIVAT10N
                         SPECIES TISSUE
                                          TA-1935

                                           1    2
                                    TA-153T

                                     1    2
                                                                 PLATE

                                                                   T»-90

                                                                    1    2
                                           TA-100

                                            I    2
       SOLVENT CONTROL
       PCSITIVE CONTROL**
UL
VL
                    12   11
                   «90 to;*
                                                    6    9
                                                  191  225
CO
O
       TEST C
            10.000000
            23.000000
            SO.OOOOOO UL
           100.000000 UL
           200.000000 UL

       ACTIVATION
       SOLVENT CONTROL     RAT
       POSITIVE CONTROL**• RAT
TEST CO  	
     10.000000 UL   RAT
     29.000000 Ul   RAT
     SO.000000 UL   RAT
    100.000000 UL   RAT
    200.000000 UL   RAT
                                   22
                                   34
                                   IT
                                   21
                                   24
21
24
21
23
20
10
 9
10
10
14
10
 «
 3
 R
 9
            LIVER    9    9
            LIVER  110  1*0
                                                    9    9
                                                  114  161
                           22   10
                          861  T2T
28   21
30   71
30   44
98   30
63   33
                           14   15
                          900 1066
                               ICO   193
                              1303  12T4
195  141
I«9  IS?
173  152
IT*  140
153  14*
                               190   190
                              1942  1T46
                                  LIVEN
                                  LIVER
                                  LIVER
                                  LIVER
                                  LIVEN
                    11
                     6
                    19
10
10
10
II
14
1C
6
8
3
1
15
9
5
3
•i
32
3T
33
23
24
21
29
1*
29
24
191
152
145
193
223
110
135
152
101
22 R
          TA-S9S9  SODIUM AZIOE
          TA-193T  9-ANINOACRIOIKE
          TA-90    2-NITROFLUORCNE
          TA-100   SODIUM AZIOE
          SOLVENT 200 UL/PLATE
                                  10 US/PLATE
                                  50 Uf/PUTE
                                  10 UGSPLATE
                                  10 US/PLATE
                                                                      TA-1535
                                                                      T A-1531
                                                                      TI-90
                                                                      TA-100
                                       2-ANTHRANINC
                                       2-ANTHRAHINC
                                       2-ANTHRANINE
                                       2-ANTHRANINC
                                          2.5  UO/PLATFI
                                          2.5  U6/PLATE
                                          2.5  UC/PLATE
                                          ?.5  HO/PLATE

-------
                  AMES SALMONELLA/MICROSOME PLATE ASSAY



1.   OBJECTIVE

     The objective of this study was to evaluate a test material for mutagenic
     activity in a bacterial assay with and without a mammalian S9 activation
     system.


2.   RATIONALE

     The Salmonella typhimurium strains used at LBI are all histidine
     auxotrophs by virtue of mutations in the histidine operon.   When
     these histidine-dependent cells are grown in a minimal media petri
     plate containing a trace of histidine, only those cells that revert
     to histidine independence (his*) are able to form colonies.   The
     trace amount of histidine allows all the plated bacteria to undergo
     a few divisions; this growth is essential for mutagenesis to occur.
     The hi£+ revertants are easily scored as colonies against the slight
     background growth.  The spontaneous mutation frequency of each strain
     is relatively constant, but when a mutagen is added to the agar the
     mutation frequency is increased 2- to 100-fold.   Cells which grow to
     form colonies on the minimal media petri plates are therefore assumed
     to have reverted, either spontaneously or by the action of a test
     substance to his+ genotype.


3.   MATERIALS

     A.   Indicator Microorganisms

     The Salmonella typhimurium strains.used  in  this  assay were  obtained
     from Dr. Bruce Ames, University of  California at Berkeley.1'5  The
     following 4 strains  were used:
Strain
Designation
TA-1535
Gene
Affected
his S
Additional Mutations
Repair
A uvr B
LPS R Factor
rfa
Mutation Type
Detected
Base-pair
substitution
     TA-1537          his  C       A uvr B     rfa        -         Frameshift

     TA-98           his  D       AuvrB     rfa     pKMIOl       Frameshift

     TA  inn          his  G       A uvr B     rfa     pKMTOl.       Base pair
      IA  IUU          	           	                            substitution
                                 131

-------
3.   MATERIALS (Continued)

     The aforementioned strains have, in addition to the mutation in the
     histidine operon, a mutation (rfa-) that leads to defective 1'POpoly-
     saccharide coat, a deletion that covers genes involved 1n the synthesis
     of vitamin biotln (bio-) and in the repair of ultraviolet (uv) - in-
     duced DNA damage (uvrB-).  The rfa- mutation makes the strains more
     permeable to many large molecules.  The uvrB- mutation decreases repair
     of some types of chemically or physically damaged DNA and thereby
     enhances the strain's sensitivity to some mutagenic agents.  The
     resistant transfer factor plasmid (R factor) pKMlOl in TA-98 and
     TA-100 is believed to cause an increase in error-prone DNA repair
     that leads to many more mutations for a given dose of most mutagens5.
     In addition, plasmid pKMlOl confers resistance to the antibiotic
     ampicillin, which is a convenient marker to detect the presence of
     plasmid in the cells.

     All indicator strains are kept at 4°C on minimal medium plates supple-
     mented with a trace of biotin and an excess of histidine.  The plates
     with plasmid-carrying strains contain in addition ampicillin (25 pg/ml)
     to ensure stable maintenance of plasmid pKMlOl.  New stock culture
     plates are made as often as necessary from frozen master cultures
     or from single colony reisolates that were checked for their genotypic
     characteristics (his, rfa. uvrB. bio) and for the presence of plasmid.
     For each experiment, an  inoculum from the stock culture plates is
     grown overnight at 37°C in nutrient broth (Oxoid CM67).

     B.   Media

     The bacterial strains were cultured in Oxoid Media #2 (nutrient Broth).
     The selective medium was Vogel Bonner Medium E with 2% glucose7.  The
     overlay agar consisted of 0.6% purified agar with 0.5 mM histidine,
     0.05 mM biotin and 0.1 M NaCl according to the methods of Ames et. al..6

     C.   Activation System

          (1)  59 Homogenate

               A 9,000 x s supernatant prepared from Sprague-Dawley adult
               male  rat liver  induced by Aroclor 1254  (described by Ames
               et. al..6) was  purchased from Bionetics  Laboratory Products,
               Litton Bionetics, Inc. and used in this  assay.
          (2)  59 Mix

                                            Concentration  per Mi Hi liter
                 Components                             S9  Mix

             NADP (sodium  salt)                         4 umoles
           D-glucose-6-phosphate                        5 umoles
                   MgCl2                                 8 umoles
                    KC1                                 33 umoles
           Sodium phosphate buffer
                   pH 7.4                             100 umoles
           Organ homogenate from rat
              liver (S9 fraction)                     100 uliters
                                  132

-------
EXPERIMENTAL DESIGN

A.   Dosage Selection

All tests are run at a minimum of four concentrations.   In the
Standard EPA Level  I Ames  assays, five dose levels of the test
material, dissolved in a suitable solvent,  are added to the test
system.  The standard test doses  for the extracted material are
10, 25, 50, 100 and 200 yliters per plate.   The solids  are tested
up to 5 mg per plate and at lower concentrations of 2.5, 1, 0.5,
0.1 and 0.05 mg per plate.   The samples are retested over a
narrower range of concentrations  with strains  showing positive
results if there is enough sample.
B.   Mutagenicitv Testing

The procedure used is based on the paper published  by Ames et. al.6
and is performed as follows:

     (1)  Nonactivation Assay

     To a sterile 13 x 100 mm test tube placed in a 43°C water bath
     the following is added in order:

          (a)  2.00 ml of 0.6% agar containing 0.05 mM histidine
                    and 0.05 mM biotin.

          (b)  0.05 ml of a solution of the test chemical to give
                    the appropriate dose.

          (c)  0.1 ml - 0.2 ml.of indicator organism(s).

          (d)  0.50 ml of 0.2M phosphate buffer, pH 7.4.

     This mixture is swirled gently and then poured onto minimal
     agar plates (see 3B, Media).   After the top agar has set, the
     plates are incubated at 37°C for approximately 2 days.  The
     number of his+ revertant colonies growing on the plates is
     counted and recorded.

     (2)  Activation Assay

     The activation assay is run concurrently with  the nonactiva-
     tion assay.   The only difference is the addition of 0.5 ml of
     S9 mix (see 3C:2, Activation System)  to the tubes in place of
     0.5 ml of phosphate buffer which is added in nonactivation
     assays.   All other details are similar to the  procedure for
     nonactivation assays.

     A detailed flow diagram for the plate incorporation assay is
     provided in Figure 1.
                           133

-------
                                   FIGURE 1

                          REVERSE MUTATION ASSAY
                          [Agar Incorporation  Method]
Aliquot of  0.5 ml
buffer
  I
-S9
                          Molten [43 to 45°C] overlay agar
                          appropriately supplemented
                                         0.05 ml
                                         <
                                          0.1 ml to 0.2 ml
                                                       ]
                                      T
                                           Test article, positive
                                           control or solvent control

                                           Aliquot of an overnight
                                           culture of bacteria
0.5 ml  S9 mix [hepatic
•*—   homogenate from
        PCS pretreated rat
        plus necessary
        cof actors]
                           Overlay poured on selective
                           bottom agar medium


                Plates incubated at 37*C for approximately 2 days
                     Number of revertants per plate counted

                                       I
                        Data entered onto preprinted forms

                            Interpretation/conclusion
                                          134

-------
4.   EXPERIMENTAL DESIGN (Continued)

     C.   Control Compounds

     A negative control consisting of the solvent used for the test
     material  1s performed 1n all cases.  For negative controls, step
     'b1 of Nonactlvatlon Assays Is replaced by 0.05 ml of the solvent.
     The negative controls are employed for each Indicator strain and
     are performed  In the absence and presence of S9 mix.   The solvent
     used to prepare the stock solution of the test material 1s given
     1n the Results section of this report.  All dilutions of the test
     material  are made  using this solvent.  The amount of solvent used
     1s equal  to the maximum volume used to give the appropriate test
     dose.

     Specific  positive  control compounds known to revert each strain are
     also used 1n the assays.  The concentrations and specificities of
     these compounds to specific strains are given In the following table:
                                              Concentration
                                                per Plate    Salmonella
        Assay        Chemical      Solvent          (yg)       strains
  Nonactlvatlon  Sodium azlde     Water            1        TA-1535, TA-100
                  2-Nitrofluorene  Dimethyl-       10        TA-98
                    (NF)             sulfoxide
                  9-aminoacridine  Ethanol         50        TA-1537
                    (9AA)

  Activation     2-anthramine     Dimethyl-        2.5      For all strains
                    (ANTH)           sulfoxlde
                                 135

-------
5.    EVALUATION CRITERIA

     Statistical methods are not currently used and evaluation is based
     on the criteria included in this protocol.

     Plate test data consist of direct revertant colony counts obtained
     from a set of selective agar plates seeded with populations of mutant
     cells suspended in a semisol id overlay.   Because the test material
     and the cells are incubated in the overlay for approximately 2 days
     and a few cell divisions occur during the incubation period, the
     test is semiquantitative in nature.  Although these features reduce
     the quantitation of result, they provide certain advantages not con-
     tained in a quantitative suspension test:

               The small number of cell divisions permits potential mutagens
               to act on replicating DNA, which is often more sensitive
               than nonreplieating DNA.

               The combined incubation of the test article and the cells
               in the overlay permits constant exposure of the indicator
               cells for approximately 2 days.

     A.   Surviving Populations

     Plate test procedures do not permit exact quantitation of the number
     of cells surviving chemical treatment.  At low concentrations of
     the test material, the surviving population on the treatment plates
     is essentially the same as that on the negative control plate.  At
     high concentrations, the surviving population is usually reduced by
     some fraction.  Our protocol will normally employ several doses
     ranging over two or three log concentrations.  This does not apply
     to spot tests and tests performed on fabrics and like materials which
     are tested at a single concentration.

     B.   Dose-Response Phenomena                i

     The demonstration of dose-related increases in mutant counts is an
     important criterion in establishing mutagenicity.  A factor that
     might modify dose-response results for a mutagen would be the selec-
     tion of doses that are too low (usually mutagenicity and toxicity
     are related).  If the highest dose is far lower than a toxic concen-
     tration, no increases may be observed over the dose range selected.
     Conversely, if the lowest dose employed is highly cytotoxic, the
     test material may kill any mutants that are induced, and the test
     material will not appear to be mutagenic.
                                 136

-------
5.   EVALUATION CRITERIA (Continued)

     C.   Control Tests

     Positive and negative control assays are conducted with each experi-
     ment and consist of direct-acting mutagens for nonactivation assays
     and mutagens requiring metabolic biotransformation in activation
     assays.  Negative controls constst of the test material solvent in
     the overlay agar together with the other essential components.  The
     negative control plate for each strain gives a reference point to
    • which the test data is compared.  The positive control assay is con-
     ducted to demonstrate that the test systems are functional with known
     mutagens.

     D.   Evaluation Criteria for Ames Assay

     Because the procedures used to evaluate the mutagenicity of the test
     material are semiquantitative, the criteria used to determine positive
     effects are inherently subjective and are based primarily on a his-
     torical data base.  Most data sets are evaluated using the following
     criteria:

          (1)  Strains TA-1535, TA-1537

          If the solvent control value is within the normal range, a test
          material producing a positive response equal to three times
          the solvent control value is considered mutagenic.

          (2)  Strains TA-98 and TA-100

          If the solvent control value is within the normal range, a test
          material producing a positive response equal to twice the solvent
          control value for TA-98 and TA-100 is considered mutagenic.

          (3)  Pattern

          Because TA-1535 and TA-100 are both derived from the same parental
          strain (6-46) and because TA-1538 and TA-98 are both derived
          from the same parental strain (D3052), to some extent there is
          a built-in redundancy in the microbial assay.  In general, the
          two strains of a set respond to the same mutagen and such a
          pattern is sought.  Generally, if a strain responds to a mutagen
          in nonactivation tests, it will do so in activation tests.

          (4)  Reproducibility

          If a test material produces a response in a single test which
          cannot be reproduced in additional runs, the Initial positive
          test data lose significance.
                                 137

-------
5.   EVALUATION CRITERIA (Continued)

     E..    Evaluation Criteria for Toxlclty

          (1)  Complete toxicity

          When there are no revertants observed on the plate(s) treated
          with the test compound, the test compound 1s defined as toxic
          to all or any of the Indicator strains at that (those) particular
          dose(s).

          (2)  Slight toxicity

          When there are fifty or less percent revertants on the plate(s)
          treated with the test compound as compared to the solvent control
          plate(s), the test compound is defined as slightly toxic to
          all or any of the indicator strains at that (those) particular
          dose(s).

     F.    Relation Between Mutagenicity and Cardnogenicity

     It must be emphasized that the Ames Salmonella/Microsome Plate Assay
     is not a definitive test for chemical  carcinogens.  It is recognized,
     howerver, that correlative and functional  relations have been demon-
     strated between these two endpoints.   The  results of comparative
     tests on 300 chemicals by McCann et.  aJL x  show an extremely good
     correlation between results of microbial mutagenesis tests and jrj
     vivo rodent carcinogenesis assays.

     All evaluations and interpretation of  the  data to be presented in
     the final report will  be based only on the demonstration, or lack,
     of mutagenic activity.
                                138

-------
                               REFERENCES
1.   McCann,J., Choi, E., Yamasaki,  E. and Ames,  B.N.:  Detection of
     carcinogens as mutagens  in the  Salmonella/microsome  test:  Assay
     of 300 chemicals.   Proc. Nat. Acad.  Sci.  USA, 72:5135-5139, 1975.

2.   Ames, B.N., Gurney, E.G., Miller, J.A.  and Bartsch,  H.:  Carcinogens
     as frameshift mutagens:  Metabolites and  derivatives of  2-acetyl-
     aminof1uorene and  other  aromatic  amine  carcinogens.   Proc. Nat. Acad.
     Sci. USA,  69:3128-3132,  1972.

3.   Ames, B.N, Lee,  F.D. and Durston, W.E.:   An  improved bacterial
     test system for  the detection and classification  of*.mutagens and
     carcinogens.  Proc. Nat. Acad.  Sci.  USA,  70:782-786, 1973.

4.   Ames, B.N, Durston, W.E., Yamasaki,  E.  and Lee, F.D.:  Carcinogens
     are mutagens:  A simple  test system  combining liver  homogenates
     for activation and bacteria  for detection.   Proc.  Nat. Acad. Sci.
     USA, 70:2281-2285, 1973.

5.   McCann,  J., Springarn, N.E., Kobori, J. and  Ames,  B.N.:  Detection of
     carcinogens as mutagens:  Bacterial  tester strains with  R  factor
     plasmids.  Proc.  Nat.  Acad.  Sci.  USA, 72:979-983,  1975.

6.   Ames, B.N., McCann, J. and Yamasaki, E.:  Methods for detecting
     carcinogens and  mutagens with the Salmonella/mammalian-microsome
     mutagenicity  test. Mutation Res.,  31:347-364,  1975.

7.   Vogel,  H.J. and  Bonner,  D.M.:  Acetylornithinase  off. col i; Partial
     purification  and some  properties.   J. Biol.  Chem., 218:97-106, 1956.
                                 139

-------
                      Q.A. Inspection Statement
                   (reference 21  CFR 58.35(b)(7))
PROJECT
                                              LBX Assay No. S3 9?
TYPE of STUDY       ftwrs ~PI*
-------
                      GENETICS ASSAY NO.  5394
                                         •^•••••OT

                          LBI SAFETY NO.  6378
    CYTQTOXIC  EVALUATION OF
       SAMPLE  f 1452
(WATER TANK DRAIN,  END OF  TEST)

            IN THE
       RODENTCELL  (CHO)
     CLONAL TOXICITY ASSAY
         FINAL REPORT
        SUBMITTED TO:

      ACUREX CORPORATION
       485 CLYDE AVENUE
   MOUNTAIN VIEW, CA  94042
        SUBMITTED BY:

    LITTON BIONETICS,  INC.
      5516 NICHOLSON LANE
  KENSINGTON, MARYLAND  20795

     LBI PROJECT NO. 20933

 REPORT DATE: DECEMBER 1980


       141

-------
                               PREFACE

This report contains a summary of the data compiled during the evaluation
of the test compound.  The report is organized to present the results in
a concise and easily Interpretable manner.  The first part contains Items
I - IX.  Items I - IV provide sponsor and compound identification infor-
mation, type of assay, and the assay design reference number.  All assay
design references indicate a standard procedure described in the Litton
Bionetics, Inc. "Screening Program for the Identification of Potential
Mutagens and Carcinogens."  Item V provides the initiation and completion
dates for the study, and Item VI provides identification of supervisory
personnel.  Item VII identifies the tables and figures containing the data
used by the study director in interpreting the test results.  The interpre-
tation itself is in Item VIII.  Item IX provides the conclusion and evalua-
tion.

The second part of the report, entitled Assay Design, describes the
materials and procedures employed in conducting the assay.  This part
of the report also contains any appendices, as well as evaluation
criteria used by the study director.  The evaluation criteria are
included to acquaint the sponsor with the methods used to develop and
analyze the test results.

All test and control results presented in this report are supported by
fully documented raw data which are permanently maintained in the files
of the Department of Genetics and Cell Biology or in the archives of
Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington, Maryland, 20795.

Copies of raw data will be supplied to the sponsor upon request.
                                 142

-------
  I.        SPONSOR:  Acurex Corporation

 II.        MATERIAL (TEST COMPOUND):  GENETICS ASSAY NUMBER:  5394

            A.   Identification:  Sample #1452 (Man Test 5/21/80, #5 Gal,    .
                                                Water Tank Drain, End of Test)
            B.   Date Received: October 6, 1980

            C.   Physical Description:  Clear, colorless liquid

III.        TYPE OF ASSAY:  Rodent Cell (CHO) Clonal Toxicity Assay

 IV.        ASSAY DESIGN NUMBER:  442

  V.        STUDY DATES:

            A.   Initiation:  November 19, 1980

            B.   Completion:  December 1, 1980

 VI.        SUPERVISORY PERSONNEL:

            A.   Study  Director:  Brian C. Myhr, Ph.D.

            B.   Laboratory Supervisor:   Robert Young, M.S.

 VII.        RESULTS:

            The data are presented  in Table  1 on page  3  and  in Figure 1
            on page 4.

VIII.         INTERPRETATION OF RESULTS:

             The test material, Sample No. 1452, was supplied as  a  colorless,
             aqueous solution that did not contain  any obvious  suspended
             material.  A portion of the sample (18 ml) was  combined with
             10X F10 medium and supplements to yield 30 ml of culture
             medium containing the test material  at a concentration of
             600 yl/ml.  This was the highest concentration  applied to the
             CHO cells.  Lower concentrations were  achieved  by  serial dilu-
             tion with F10 culture medium.  The treatments were initiated
             by replacing the F12 medium in the cell cultures with  F10
             SdlS containing the test material.   After the 24 hour expo-
             suVe period, the cultures were returned to Fl2  medium and
             incubated for colony development.  The test material did not
             cause any visible precipitation of culture medium components,
             but I thift to a slightly acidic PH of 6.26 was observed for
             the 600 yl/ml concentration.  The pH remained normal (above
             7.0) for the other test concentrations.
                                      143

-------
VIII.  INTERPRETATION OF RESULTS:   (continued)

       Two trials of the assay were performed.  The first trial  was
       considered insufficient for evaluation because the average
       cloning efficiency of the negative controls was 67.4%, which
       was slightly below the 70% criterion used for an acceptable
       assay.  The results of the second trial are presented in  Table
       1 and Figure 1 .

       Seven test concentrations from 6 yl/ml to 600 yl/ml  were
       evaluated for their effects on colony survival.  As  shown in
       Table 1, complete lethality was obtained for treatments in the 60
       to 600 yl/ml concentration range.  In contrast, the  treatments
       with 6 to 30 vl/ml were essentially nontoxic to the  cells.  Thus,
       an extremely sharp survival curve was obtained, as shown  in Figure
       1, with an EC50 apparently located at 45 yl/ml.  [In the  first trial
       (results not shown), an EC50 value of 52 yl/ml  was obtained for a
       somewhat broader survival curve].  The EC50 was therefore located
       at the upper end of the 6-60 yl/ml concentration range, which is
       the range defined for moderate toxicity for aqueous  liquids (see
       Evaluation Criteria).  Since complete lethality was  obtained at the
       boundary between moderate and low toxicity, an evaluation of moderate
       toxicity to CHO cells appeared appropriate.

  IX.  CONCLUSIONS:

       The test material, Sample No. 1452, was evaluated as having moderate
       toxicity in the CHO Clonal  Toxicity Assay.   The EC50 value was
       45 yl/ml.

                                         Submitted by:

                                         Study Director:
                                                  C. ftk        13,/3/fr?
                                         Brian C.  Myhr, PKjD.       Date
                                         Section Chief
                                         Mammalian Genetics
                                         Department of  Genetics
                                         and Cell  Biology
                                         Reviewed by:
                                         David J. /Brusick, Ph.D.   Date
                                         Director
                                         Department of Genetics
                                         and Cell  Biology
                                      144

-------
                                     TABLE  1
                     RODENT CELL (CHO) CLONAL TOXICITY ASSAY
Sample Identity:   Sample 11452r	
Tank Drain. End of Test)	
Description of Sample:  Clear, colorless
aqueous liquid	
LBI Assay No.:   5394	
                                          EC50 Value:    45 ,,i/mi
                                          Toxicity
                                          Classification:  Moderate
Date Received:   October 6, 1980	
Test Date:   November 25. 1980 (Trial 2)
Vehicle:    None	
Cell Type:   CHO-K1	
Cells Seeded per Dish:   200	
                                          pH Alterations:  pH 6.26 at 600 n
                                          Comments on Treatment:  Normal DH
                                           for treatments with 6 to 300 ul/ml
                              CLONAL TOXICITY DATA
Sample
NC
Test
Test
Test
Test
Test
Test
Test
Applied
Concentration
yl/ml
—
6.0
10.0
30.0
60.0
100.0
300.0
600.0
Dish
#1
187
190
214
167
0
0
0
0
Dish
#2
193
219
210
190
0
0
0
0
Dish
#3
187
183
181
178
0
0
0
0
Average
Count
189.0
197.3
201.7
178.3
0
0
0
0
Relative
Survival
%
100.0
104.4
106.7
94.3
0
0
0
0
Cloning
Efficiency
%
94.5







 NC = Negative Control,  F12  medium.
                                        145

-------
                    FIGURE 1

       RODENT CELL  (CHO) CLONAL TOXICITY ASSAY
                EC50 DETERMINATION

SAMPLE #1452   (WATER TANK  DRAIN,  END OF  TEST)
            10
1000
                CONCENTRATION, pi/ml
                     146

-------
                             ASSAY DESIGN NO. 442

                    RODENT CELL CLONAL  (CHO) TOXICITY ASSAY


This assay conforms to one of the bioassay tests in the US Environmental Protec-
tion Agency (EPA) IERL - RTP Procedures Manual:  Level 1 Environmental Assessment
Biological Tests (September, 1980).   Level 1 bioassays obtain preliminary infor-
•ation on the harmful biological effects of chemicals found in industrial feed
and waste streams by using mammalian  cells in culture to measure metabolic
impairment and cellular death.


1.   OBJECTIVE

     The objective of this assay is to  determine the concentration of a test
     article that will reduce the colony forming ability of Chinese hamster
     cells by 50% after a 24-hour exposure.  This concentration is referred to
     as the EC50 value.


2.   MATERIALS

     A.   Indicator Cells

          The indicator cells used in this study are Chinese hamster CHO-K1
          cells (American Type Culture  Collection No. CCL 61).  This cell type
          was derived from ovarian tissue and has spontaneously transformed to
          a stable, hypodiploid line  of rounded, fibroblastic cells with un-
          limited growth potential.   Monolayer cultures have a fast doubling
          time of 11 to 14 hours, and untreated cells can normally be cloned
          with an efficiency of 80% or  greater.  Permanent stocks are main-
          tained in liquid nitrogen and laboratory cultures are maintained by
          serial subculturing.  Laboratory cultures are periodically checked
          by culturing methods for the  absence of mycoplasma contamination.
          This test system is specified by the IERL- RTP Procedures Manual.

     B.   Medium

          The CHO-K1 cell line has an absolute requirement for proline and
          therefore must be maintained  in a culture medium containing suf-
          ficient amounts of this amino acid.  Ham's F12 medium, which contains
          3 x 10-4M L-proline, is used, supplemented with 10% fetal bovine
          serum, 100 units per milliliter of penicillin, 100 ug streptomycin
          per milliliter, and 0.5 pg  amphotericin B (Fungizone) per milliliter.

     C.   Controls

          Untreated cells are cloned  to establish the control cloning effi-
          ciency.  If the test article  is dissolved in an organic solvent
          (usually dimethylsulfoxide),  cells exposed to solvent fn the medium
                                      147

-------
2.   MATERIALS (Continued)

          are cloned to provide the reference cloning efficiency for the effect
          of the test article.  The final concentration of solvent in the growth
          medium is generally IX or less.  All controls are performed in tripli-
          cate.

     D.   Sample Forms

          Solid samples are tested as a solution or are ground to fine particles
          (less than 5 urn) and tested as a suspension in growth medium.  Dry
          particulate articles, aqueous liquids, suspensions and slurries are
          added directly to the growth medium and tested as a suspension or
          solution.  Liquids containing less than 0.2% organic solvent are
          generally tested directly; samples dissolved in organic solvents are
          solvent exchanged in dimethylsulfoxide (DMSO) before testing.  Original
          sample volumes may be reduced a maximum of 10-fold during solvent
          exchange, and the concentration factor is used to convert assayed
          volumes into equivalent original sample volumes.  All sample manipu-
          lations are performed as described in the IERL-RTP Level 1 Procedures
          Manual.


3.   EXPERIMENTAL DESIGN

     A.   Dose Selection

          Unless the approximate toxicity is already known or the sample size
          is  limiting, the following dose ranges are tested for different sample
          forms.  Dry particulate articles are dissolved or suspended  in growth
          medium and tested at five dose levels from 1000 ug/ml to 10  ug/ml.
          Aqueous samples, suspensions or slurries are tested from 600 uliters/ml
          to  6 uliters/ml in  five dose steps.  Samples that are solvent exchanged
          into DMSO are tested from 20 uliters/ml to 0.2 uliters/ml, also in five
          dose steps.  Solvent concentrations are IX for all dose levels except
          the maximum applicable dose (MAD) of 20 ul/ml, which contains 2%  solvent.
          A second dose study is performed with an appropriate dose  range if the
          EC50 has not been properly located  in the initial test; EC50 values
          greater than 1,000  ug/ml, 600  uliters of aqueous  sample/ml or 20  uliters
          nonaqueous sample/ml are not determined.

     B.   Clonal Toxicity Assay

          Cells  from monolayer stock cultures in  logarithmic growth  phase are
          trypsinized, counted by hemacytometer and reseeded into a  series  of
          60- or 100-mm culture dishes at 200 cells per dish.  The cultures
          are incubated for 6 to 16 hours at  37°C to  allow attachment  of the
          cells  and recovery  of growth rate.

          Test article is then applied (three dishes  per dose),  and the cultures
          returned to the incubator.  If the  test article causes a  color change,
          an additional dish  is treated  with  the  high dose and the  pH of the


                                      148

-------
3.    EXPERIMENTAL DESIGN (Continued)

          medium Is determined.  The pH is also recorded for the highest dose
          which results in a slight color change.  After a 24-hour exposure
          period the medium is aspirated and the cells washed with Dulbecco's
          phosphate buffered saline (PBS; prewarmed to 37°C).  The pH of the
          discarded medium for which initial pH measurements were made is again
          recorded.  Fresh medium  is placed on each culture and incubation con-
          tinued for an additional 6 days to allow colony development.  Medium
          is drained from the cultures and the surviving colonies are washed
          with PBS, fixed in methanol, and stained with Giemsa.  Colony counting
          is performed by eye; colonies smaller than 50 to 100 cells are not
          counted.
4.   ASSAY ACCEPTANCE  CRITERIA

     The assay  is  considered acceptable  for evaluation of the test results if
     the following criteria are  met:

                The average cloning efficiency  of the CHO-K1 cells in the nega-
                tive controls is  70% or greater, but not exceeding 115%.

                The distribution  of colonies in the treated cultures is generally
                uniform over the  surface  of the culture dish.

                The data points for each  test concentration critical to the loca-
                tion of the EC50  are .the  averages of at least two treated cultures.

                A sufficient number of test concentrations are available to
                clearly locate the EC50 within  a toxicity region as defined
                under Assay Evaluation Criteria.

                If the EC50 value is greater than 1000 M9/">1, 600 jjliters of
                aqueous sample/ml, or 20  uliters of nonaqueous sample/ml, the
                plotted curve does not exceed 110% of the negative control.


 5.   ASSAY EVALUATION CRITERIA

     The  screened doses, pH values (if appropriate), colony  counts, percent
     survivals  (colony counts relative to control colony counts) and  EC50
     values are provided.  The percent survival  is plotted as a function of
     applied concentration and the EC50 value  determined graphically  by fitting
     a curve by eye through the data points.   The EC50  is used to  rank the test
     •aterial using the standard evaluation  criteria defined in the table below.

     Sorbent extracts of known organic content are  evaluated as nonaqueous
     liquids and in terms of their solid contents  (pg  orgamcs/ml), and are
     ranked using the more sensitive parameter.  Where  data  on the original
     liquid or gas sample volumes is provided by the  Sponsor the-EC50 value
     is calculated in terms of these equivalent volumes per  mill11 ter of
     culture medium; however, no evaluations are made  as yet on this  basis.


                                        149

-------
ASSAY EVALUATION CRITERIA (Continued)
   Toxicity3
     Solids
(EC50 in ug/ml)
Aqueous Liquids
(EC50 in Ml/ml)
Nonaqueous Liquids
 (EC50 in Ml/ml)
High
Moderate
Low
Not Detectable
10 to 100
100 to 1000
>1000
<6
6 to 60
60 to 600
>600
.2-2
2-20
>20

 Evaluation criteria formulated by Litton Bionetics, Inc. for IERL-RTP
 Procedures Manual:  Level 1 Environmental Assessment Biological Tests.

 Criteria for nonaqueous liquids are tentative and under evaluation.


RECORDS TO BE MAINTAINED

All raw data, protocols, protocol modifications, test article weight and
dispensation records and correspondence between LBI and the Sponsor are
being maintained in a central file within the Department of Genetics and
Cell Biology.  These records will be filed under departmental assay number
and held up to 2 years following submission of the final report to the
Sponsor.  After 2 years they will be transferred to the LBI archives for
permanent storage.
                                 150

-------
                      Q.A.  Inspection Statement
                   (reference 21 CFR 58.35(b)(7))
                                              LSI Assay No.
TYPE Of STUDY  £&.&!+> OtJiQ. (
                                                           J   i^LOuLJ
         This final study report was reviewed by the LBI  Quality
Assurance Unit on  /Qi. u^3
         The short-term nature' of this study precluded inspection while
it was in process.  The Quality Assurance Unit inspects an in-process
study of this type approximately once per month to assure that no
significant problems exist that are likely to affect the integrity of
this type of study.
                                      Auditor, Quality Assurance Unit
                                     151

-------
                     GENETICS ASSAY NO.     5394
                          LBI SAFETY NO.    6378
 MUTAGENICITY EVALUATION  OF
        SAMPLE  #1452
           IN THE
 AMES SALMONEIEATMICROSOME
        PLATE TEST
       FINAL REPORT
       SUBMITTED TO:
   ACUREX CORPORATION
    485 CLYDE AVENUE
MOUNTAIN VIEW, CA  94042
      SUBMITTED BY:
 LITTON BIONETICS, INC.
  5516 NICHOLSON LANE
KENSINGTON, MARYLAND  20795
  LBI PROJECT NO. 20988
  REPORT DATE;  DECEMBER,  1980

            152

-------
                         PREFACE
This report contains a summary of the data compiled during the
evaluation of the test compound.  The report is organized to
present the results in a concise and easily interpretable manner.
The first part contains items I-IX.  Items I-IV provide sponsor
and compound identification information, type of assay, and the
protocol reference number.  All protocol references indicate a
standard procedure described in the Litton Bionetics, Inc.
"Screening Program for the Identification of Potential Mutagens
and Carcinogens."  Item V provides the initiation and completion
dates for the study, and Item VI provides identification of super-
visory personnel.  Item VII identifies the tables and/or figures
containing the data used by the study director in interpreting
the test results.  The interpretation itself is in Item VIII.
Item IX provides the conclusion and evaluation.

The second part of the report describes the materials and procedures
employed in conducting the assay.  This part of the report also
contains evaluation criteria used by the study director, and any
appendices.  The evaluation criteria are included to acquaint the
sponsor with the methods used to develop and analyze the test
results.

All test and control results presented in this report are supported
by fully documented raw data which are permanently maintained in
the files of the Department of Genetics and Cell Biology or in the
archives of Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington,
Maryland  20795.
                              153

-------
   I.   SPONSOR:  Acurex Corporation

  II.   MATERIAL  (TEST COMPOUND):    GENETIC ASSAY NUMBER: 5394

       A.   Identification: Sample Number 1452 and its Extract

       B.   Date  Received  : October  6, 1980

       C.   Physical  Description: Neat Sample: Colorless Liquid
                                 Extract  : Green Liquid

 III.   TYPE OF  ASSAY :  Ames Salmonella/microsome Mutagenesis Assay

  IV.   PROTOCOL NUMBER: 401 (EPA-LEVEL  I)

   V.   STUDY DATES:

       A.   Initiation:  November  21, 1980

       B.   Completion:  November  25, 1980

  VI.   SUPERVISORY PERSONNEL:

       A.  Study Director:  D.R. Jagannath, Ph.D.

 VII.    RESULTS:

       The results of this are presented  in  Tables  1  and 2.

VIII.    INTERPRETATION OF  RESULTS:

        The test material, Sample No. 1452 was received  as  a  clear colorless
        liquid (1 gallon)  with no visible  suspended  particulates.  An aliquot
        of the neat sample (3250  ml  ) was  concentrated 500  fold  by passing
        through the XAD-2  resin column  at  a  flow rate  of 3  ml  per  minute.   The
        resin was then extracted  with methylene  chloride in a Soxhlet apparatus
        for 24 h.  The extract  (250  ml  ) was  concentrated to  approximately 5 ml
        using a Kuderna  Concentrator.  The concentrated  extract  was  then solvent
        exchanged by sequential addition of  Dimethylsulfoxide (DMSO) and evapo-
        ration of residual methylene chloride in a warm  water bath under a
        stream of nitrogen.  Final  sample  size  in DMSO was  6.5 ml  which repre-
        sents a 500 fold concentration factor.   The  concentrate  was  a clear
        solution with a  pale  blue/green color.

        The test materials, neat  sample #1452 and its  extract were examined for
        mutagenic activity in  the Ames Salmonella assay  in  the presence and
        absence of liver microsomal  enzymes  from Aroclor induced rats.

        Solvent controls,  distilled  water  for the neat sample and DMSO for the
        extract of the sample,  and  specific  positive controls were also assayed
        concurrently with  the test  materials.
                                       154

-------
         MNGE:  The  dose range employed for the neat sample was  from 10.0 ul
    to 200.0  ul  per plate and for the extract it was  from 5.0  ul to 100 0 ul
    per plate (due  to the limited amount of the extract).  The  organic content
    of the  sample was not supplied by the sponsor.

    TOXICITY; The  test materials, neat sample and  its extract  were not toxic to
    any of  the strains used in this assays.

    RESULTS:

    The results  of  the tests conducted on the test  material, neat sample, in the
    absence of an activation system was negative.   However, the extract of the
    sample  exhibited  genetic activity with the strain TA-98 in  the absence of an
    activation system.  The response was slightly above two times the background
    level and was observed at the highest concentration of 100  ul per plate.

    The results  of  the tests conducted on the test  materials neat sample and its
    extract were all  negative in the presence of rat  liver activation system.

    Dose- related increases in the number of revertants were also observed with the
    base-pair substitution strain TA-100 in  the non-activation  and activation
    assays  performed  on the extract.  However these increases did not meet the
    criteria  for a  positive response.

IX.  CONCLUSION:

    The test  material, neat sample #1452 and its extract were tested in the
    Ames/Salmonella assay and were compared  for their mutagenic activities.  The
    neat sample  did not exhibit genetic activity in any of the  assays conducted
    in this evaluation and was considered not mutagenic under these conditions.
    However,  the extract of the sample #1452 exhibited genetic  activity with
    the strain TA-98  in the nonactivation assays conducted in this evaluation
    and is  considered as mutagenic.   Based on the EPA's "Definition of Toxicity
    Categories for  Health Effects Tests" the mutagenic activity of the neat
    sample  #1452 is classified as 'ND1  and the mutagenic activity of the extract
    of the  sample #1452 is classified  as 'low'.   Since organic  content of the
    sample  was not  available the specific activity  has not been calculated.
Reviewed by:
  -
JLf
 "
Date
    Director
    Department of Genetics
    and Cell Biology
                                              Submitted by:

                                              Study  Director
                                         D.R. Jagannath, Ph.D.
                                         Section Chief
                                         Submammalian Genetics
                                         Department of Genetics
                                         and Cell Biology
                                                                      Date
                                       155

-------
           RESULTS
                                                          TAHIE I
O1
         ».    MANE  OR CODE  O'SICNATION OF  THC TEST COMPOUND:  SAHPLF §14«2 (EXTRACT)
         n.    SOLVENT:  ON2C
         c.    TCSI  INITMTIOW DATES:  u/2i/*o
         D.    TESI  COMPLETION OATr: H/It/HO
         £•    s-9 LOTI:  0*009
         NCTC:    CONCENTRATIONS  *RE 6t«CN  IN NICROLITER!   PER  PLATE
TEST SPECIES TISSUC

NOMCTIVATION
SOLVENT CONTROL .
POSITIVE CONTROL** -~
TEST COMPOUND
10.000000 UL
25.000000 UL — 	
50.000000 UL — 	
100.000000 UL 	 	
200.000000 UL
ACTIVATION
SOLVENT CONTROL HAT LIVER
POSITIVE CONTROL*** RAT LIVER
TESI COMPOUND
10.000000 Ul RAT LIVER
25.000000 UL RAT LIVER
•0.000000 UL RAT LIvrR
100.000000 UL RAT LIVER
200.000000 UL "AT LIVEN
*•
U-I53S SODIUM A/IDE
TA-153T 9-AHINOACRI01NE
IA-9R 2-NITHI)FLUORTN?
TA-IOO snoiu" tr'.ot
SOLVtNT 200UL/PLAU

TA-1531:


21 39
6?0 662

11 2«
31 :T
32 •!
29 Ifl
22 19

2T 31
163 164

2T 11
?T 19
4? 1*
2H IK
'1 14





»A-1«3» TA-98


13
TIT

R
1
16
R
Q

10
262

10
*
f.
*
T



It
84 R

11
9
^
«
12

16
210

T
1
1.1
t
*



2«
1013

20
3«
22
79
33

46
130?

16
79
?f
2«
16
* * *


21
*90

??
20
??
31
70

•in
1263

2M
3'
tT
•>*,
74



TA-IOA
3*
I
126
T01

142
146
115
12R
139

145
1505

ion
TT
RT
•iy
106

10 UI5/PHT* T«-tS3« 2-ANTMl


sp \f.
tO IJG
/PI 'T'
/PLA^f


10 l|f/PL»Tr




T»-11ST
T A-"R
TA-109

2t(
5
1T6
654

119
142
11?
150
12«

115
1«21

TR
91;
RT
It.
9-»

•AMI NT 2.5 UC/PLATr
7-tKTH'AMMC J.5 U«/PlATr
?*I*JTHI
1AMINC ?.5 UH/PLAf
7-ANTHPAMINr 2.« UG/PLATf



-------
                     NAPE OP CODE OFSICNATION OP THE TEST COMPOUND:  SAMPLE
                     SOLVENT:  ONSO
                     TEST INITIATION DATES:  ll'2!'4n
                     TEST COMPLETION DATE: ll/2*/HO
                     S-9 LOTt: 01009
                       CONCENTRATIONS ARE SIVEN IN MICRCLITER*   PER  PLATE
A.
B.  I
C.
0.
E.
NOTE:
               TEST
               NCNACTIVATION
                                                                                   (EXTRACT)
                                 SPECIES TISSUE
                                                  1A-153S

                                                   1    ?
                                                    TI-153T

                                                     t     ?
                           L A T f


                           TA-98

                            1    f
                                TA-10J

                                 1    2
SCLVEkT CONTROL

POSITIVE CONTROL**

TEST  CORPOUN1
       5.000000 UL
      10.000000 UL
      25.000000 UL
      50.000000 UL
    100.000000 UL

ACTIVATICk
                                                  32    35

                                                 620  6*2
            T    T

          797  84ft
                                                   II
                                                   IT
                                                   20
                                                   23
19
22
!2
23
10
II
1?
16
1"
 4
13
                                                                        14
  31

1013


  3D

  3ft
  44
  *9
33
31
12
46
                               144  141

                               Tfll  654
164  162
1T4  1T3
lift  164
213  199
233  220
Ul
-si
               SOLVENT CONTROL      RAT

               POSITIVE  CONTROL***  RAT
                            LIVER   34   57

                            LIVER  163  1M
           1R    9

          262  210
                39

              1302
                                                                                        3T
                          153  143

                         15(« l«23
TEST COMPOUND
5.000000 UL
10.0COOOO UL
25.000006 UL
•0.000000 UL
100.000000 UL

RAT
RAT
RAT
RAT
RAT

LIVER
LIVER
LIVER
LIVER
LIVfR

42
53
26
9*i
32

42
40
3*
4«
34

7
1
13
!
-------
                                                            TABLE A
                                    DEFINITION OF TOXICITY CATEGORIES FOR HEALTH EFFECTS  ASSAYS*
en
00
Assay9
Ames

RAM

CHO

WAT


'Standard
Ames:
RAM:
CHO:
WAT:
Activity Measured6 Type5 MADd
MEC S 5
(Mutagenesls) AL.NAL 200
ECjo s !
(lethality) AL 600
NAL 20
ECSO S 1
(lethality) AL 600
NAL 20
LDSO S 5
(lethality and AL.NAL 5
toxic signs)
test abbreviations are as follows:
Ames SalMonella/nicrosone Mutagenesls assay
Rabbit alveolar macrophage cytotoxicity assay
Rodent cell clonal toxicity assay
Acute in vivo test in rodents (whole animal test)
Ranae of Concentration or Dosage
Units
ing/plate
pi/plate
mg/ml
Ml /ml
pi /ml
mg/ml
Ml/ml
Ml/ml
gm/kg
ml /kg






High
<0.05
<2
<0.01
<6
<0.2
<0.01
<6
<0.2
<0.05
<0.05






Moderate
0.05-0.5
2-20
0.01-0.1
6-60
0.2-2
0.01-0.1
6-60
0.2-2
0.05-0.5
0.5-0.5






Low Not Detectable
(ND)
0.5-5
20-200
0.1-1
60-600
2-20
0.1-1
60-600
2-20
0.5-5
0.5-5






ND at >5
ND at >200
ND at >1
ND at >600
ND at >20
ND at >1
ND at <600
ND at >20
ND at <5
ND at <5






        Standard abbreviations for measured endpoints  are as  follows:
            MEC:   Minimum effective concentration
           ECSO:   Calculated concentration expected to produce effect  in 50 percent of population
           LD50:   Calculated dose expected to kill  50  percent of population

       CS = Solid, AL = Aqueous liquid,  NAL = Nonaqueous liquid

       dMAD = Maximum applicable dose
       "Evaluation criteria formulated by Litton Bionetics, Inc.  for 1ERL-RTP
        Procedures Manual:  Level 1 Environmental Assessment BiologicaT Tests.

-------
                   AMES SALMONELLA/MTCRQSOME PLATE A«;AV



 1.    OBJECTIVE




                              assay with and without a mammalian S9 activation"
 2.    RATIONALE


      luxof?^?^                         at Ul are all histidine
                   n
      DUte  cotMln  PJ"dent  5*1  S *re 9rown 1n a m1nimal ^1. petri
      to  MitldlSi J2.I  I?" 1?1s'1d1ne. only those cells that revert
      t?ace  Liolnt I!  !?%"•;•"  CMi*) *re able to fon" "lonies.   The
                      h'?Vdlne  a11?« all the plated bacteria to  undergo
                     ;  ?hls-9fowtl? 1s essential for mutagenesis to occu?.

                          "I6 "Slly $C°red " C0lon1es a9a1nst the sll'9ht
                        *  J6 5pontaneous -utitlon frequency of each strain
                    const?nt! but when a "«"tagen is added to the agar the
                           ™™?" *- to 100-fold.   Cells which grow to
                   *       "inimal media petri P1ates  are therefore assumed
      to  have reverted,  either spontaneously or by the action of a test
      substance to  hj_s+  genotype.



3.   MATERIALS


     A.   Indicator Microorganisms


     The Salmonella typhimurium strains.used in this assay were obtained
from Dr. Bruce Ames, University of California at Berkeley.3
following 4 strains were used:
Strain
Designation
TA-1535
TA-1537
TA-98
TA-100
Gene
Affected
his G
his C
his D
his G
Additional Mutations
Repair
A uvr B
A uvr B
A uvr B
A uvr B
IPS R Factor
rfa
rfa.
rfa pKMIOl
rfa pKMlOT
l'5 The
Mutation Type
Detected
Base-pair
substitution
Frameshift
Frameshift
Base pair
                                                                  substitution
                                  159

-------
3.    MATERIALS (Continued)

     The aforementioned strains have, in addition to the mutation in the
     histidine operon, a mutation (rfa-) that leads to defective lipopoly-
     saccharide coat, a deletion that covers genes involved in the synthesis
     of vitamin biotin (bio-) and in the repair of ultraviolet (uv) - in-
     duced DNA damage (uvrB-).   The rfa- mutation makes the strains more
     permeable to many large molecules.   The uvrB- mutation decreases repair
     of some types of chemically or physically damaged DNA and thereby
     enhances the strain's sensitivity to some mutagenic agents.  The
     resistant transfer factor plasmid (R factor) pKMlOl in TA-98 and
     TA-100 is believed to cause an increase in error-prone DNA repair
     that leads to many more mutations for a given dose of most mutagens5.
     In addition, plasmid pKMlOl confers resistance to the antibiotic
     ampicillin, which is a convenient marker to detect the presence of
     plasmid in the cells.

     All indicator strains are kept at 4°C on minimal medium plates supple-
     mented with a trace of biotin and an excess of histidine.  The plates
     with plasmid-carrying strains contain in addition ampicillin (25 ug/ml)
     to ensure stable maintenance of plasmid pKMlOl.  New stock culture
     plates are made as often as necessary from frozen master cultures
     or from single colony reisolates that were checked for their genotypic
     characteristics (his, rfa. uvrB. bio) and for the presence of plasmid.
     For each experiment, an inoculum from the stock culture plates is
     grown overnight at 37°C in nutrient broth (Oxoid CM67).

     B.   Media

     The bacterial strains were cultured in Oxoid Media #2 (nutrient Broth).
     The selective medium was Vogel Bonner Medium E with 2% glucose7.   The
     overlay agar consisted of 0.6% purified agar with 0.5 mM histidine,
     0.05 mM biotin and 0.1 M Nad according to the methods of Ames et. al_.6

     C.   Activation

          (1)  S9 Homogenate

               A 9,000 x £ supernatant prepared from Sprague-Dawley adult
               male rat liver induced by Aroclor 1254 (described by Ames
               et. al_.6) was purchased from Bionetics Laboratory Products,
               Litton Bionetics, Inc. and used in this assay.
          (2)  S9 Mix

                                            Concentration per Milliliter
                 Components                            59 Mix

            NADP  (sodium  salt)                         4 umoles
          D-glucose-6-phosphate                        5 umoles
                  MgCl2                                8 umoles
                   KC1       .                          33 umoles
          Sodium  phosphate buffer
                  pH 7.4                              100 umoles
          Organ homogenate from  rat
             liver (S9 fraction)                      100 uliters

                                   160

-------
4.   EXPERIMENTAL DESIGN

     A.   Dosage Selection

     All tests are  run  at a minimum of four concentrations.   In the
     Standard EPA Level  I Ames assays, five dose levels of the test
     material, dissolved in a suitable solvent,  are added to the test
     system.  The standard test doses for the extracted material are
     10, 25, 50, 100  and 200 ^liters per plate.   The solids  are tested
     up to 5 mg per plate and at lower concentrations of 2.5, 1, 0.5,
     0.1 and 0.05 mg  per plate.  The samples are retested over a
     narrower range of  concentrations with strains  showing positive
     results if there is enough sample.
     B.   Mutaqenicitv Testing

     The procedure used is based on the paper published by  Ames et. aj.6
     and is performed as follows:

          (1)  Nonactivation Assay

          To a sterile 13 x 100 m test tube placed in a 43°C water bath
          the following is added in order:

               (a)  2.00 ml of 0.6% agar containing 0.05 mM histidine
                         and 0.05 mM biotin.

               (b)  0.05 nl of a solution of the test chemical to give
                         the appropriate dose.

               (c)  0.1 ml • 0.2 ml of indicator organism(s).

               (d)  0.50 ml of 0.2M phosphate buffer, pH 7.4.

          This mixture is swirled gently and then poured onto minimal
          agar plates (see 3B, Media).  After the top agar  has set, the
          plates are incubated at 37°C for approximately 2  days.  The
          number of his-*- revertant colonies growing on the  plates is
          counted and recorded.

          (2)  Activation Assay

          The activation assay is run concurrently with the nonactiva"
          tion assay.   The only difference is the addition  of 0.5 ml of
          S9 mix (see 3C:2, Activation System)  to the tubes in place of
          0.5 ml of phosphate buffer which is added in nonactivation
          assays.  All other details are similar to the procedure for
          nonactivation assays.

          A detailed flow diagram for the plate incorporation assay is
          provided in Figure 1.
                                 161

-------
                                   FIGURE 1

                          REVERSE MUTATION ASSAY
                          [Agar Incorporation Method]
                          Molten [43 to 45* C] overlay agar
                          appropriately supplemented
                                          0.05 ml
                                          0.1 ml to 0.2 ml
Aliquot of   0.5 ml
buffer       —
-S9
                                           Test article i positive
                                            control or solvent control

                                            Aliquot of an  overnight
                                            culture of bacteria
      0.5 mi  S9 mix [hepatic
4- SB
  **  •*—    homogenate from
              PCS pretreated rat
              plus necessary
              co factors]
                           Overlay poured on selective
                           bottom agar medium

                Plates incubated at 37* C for approximately 2 days

                     Number of revertants per plate counted

                                       \
                        Data entered onto preprinted forms

                            Interpretation /conclusion
                                          162

-------
4.   EXPERIMENTAL DESIGN (Continued)

     C.   Control Compounds
                C°nt!r01 Con!1st1n9 * the solvent used for the test
         «       perfofmedA1n a11 "ses.  For negative controls,  step
         «! N°nactlvat10n Assays is replaced by 0.05 ml of the solvent.
      *  "fSJtive controls are employed for each indicator strain and
     are performed In the absence and presence of S9 mix.   The solvent
     used to prepare the stock solution of the test material  is given
     in the Results section of this report.   All dilutions of the test
     material are made using this solvent.  The amount of  solvent used
     is equal to the maximum volume used to give the appropriate  test


     Specific positive control compounds known to revert each strain  are
     also used in the assays.   The concentrations and specificities of
     these compounds to specific strains are given in the  following table:
                                             Concentration
       A            ^                          Per  Plate     Salmonella
       Assay        Chemical      Solvent           (ug)        strains


  Nonactivation  Sodium azide     Water             1         TA-1535, TA-100
                 2-Nitrofluorene  Dimethyl-       10         TA-98
                   (NF)             sulfbxlde
                 9-aminoacridine  Ethanol         50         TA-1537
                   (9AA)

  Activation     2-anthramine     Dimethyl-         2.5       For all strains
                   (ANTH)           sulfoxide
                                 163

-------
5.   EVALUATION CRITERIA

     Statistical methods are not currently used and evaluation is based
     on the criteria included in this protocol.

     Plate test data consist of direct revertant colony counts obtained
     from a set of selective agar plates seeded with populations of mutant
     cells suspended in a semisol id overlay.  Because the test material
     and the cells are incubated in the overlay for approximately 2 days
     and a few cell divisions occur during the incubation period, the
     test is semiquantitative in nature.  Although these features reduce
     the quantitation of result, they provide certain advantages not con-
     tained in a quantitative suspension test:

               The small number of cell divisions permits potential mutagens
               to act on replicating DNA, which is often more sensitive
               than nonreplieating DNA.

               The combined incubation of the test article and the cells
               in the overlay permits constant exposure of the indicator
               cells for approximately 2 days.

     A.   Surviving Populations

     Plate test procedures do not permit exact quantitation of the number
     of cells surviving chemical treatment.  At low concentrations of
     the test material, the surviving population on the treatment plates
     is essentially the same as that on the negative control plate.  At
     high concentrations, the surviving population is usually reduced by
     some fraction.  Our protocol will normally employ several doses
     ranging over two or three log concentrations.  This does not apply
     to spot tests and tests performed on fabrics and like materials which
     are tested at a single concentration.

     B.   Dose-Response Phenomena

     The demonstration of dose-related increases in mutant counts is an
     important criterion in establishing mutagenicity.  A factor that
     might modify dose-response results for a mutagen would be the selec-
     tion of doses that are too low (usually mutagenicity and toxicity
     are related).  If the highest dose is far lower than a toxic concen-
     tration, no increases may be observed over the dose range selected.
     Conversely, if the lowest dose employed  is highly cytotoxic, the
     test material may kill any mutants that  are induced, and the test
     material will not appear to be mutagenic.
                                  164

-------
5.   EVALUATION CRITERIA  (Continued)

     C.   Control Tests

     Positive and negative  control assays are conducted with each experi-
     ment and consist of  direct-acting mutagens for nonactivation assays
     and mutagens requiring metabolic biotransformation in activation
     assays.  Negative controls  consist of the test material solvent in
     the overlay agar together with the other essential components.  The
     negative control plate for  each strain gives a reference point to
    • which the test data  is compared.  The positive control assay is con-
     ducted to demonstrate  that  the test systems are functional with known
     mutagens.

     D.   Evaluation Criteria for Ames Assay

     Because the procedures used to evaluate the mutagenicity of the test
     material are semi quantitative, the criteria used to determine positive
     effects are inherently subjective and are based primarily on a his-
     torical data base.   Most data sets are evaluated using the following
     criteria:

          (1)  Strains TA-1535,  TA-1537

          If the solvent  control value is within the normal range, a test
          material producing a positive response equal to three times
          the solvent control value is considered mutagenic.

          (2)  Strains TA-98 and TA-100

          If the solvent  control value is within the normal range, a test
          material producing a positive response equal to twice the solvent
          control value for TA-98 and TA-100 is considered mutagenic.

          (3)  Pattern

          Because TA-1535 and TA-100 are both derived from the same parental
          strain (G-46) and because TA-1538 and TA-98 are both derived
          from the same parental strain (D3052), to some extent there is
          a built-in redundancy  in the microbial assay.  In general, the
          two strains of  a  set respond to the same mutagen and such a
          pattern is sought.  Generally, if a strain responds to a mutagen
          in nonactivation  tests, it will do so in activation tests.

          (4)  Reproducibility

          If a test material produces a response in a single test which
          cannot be reproduced in additional runs, the Initial positive
          test data lose  significance.
                                  165

-------
5.   EVALUATION CRITERIA (Continued)

     E:   Evaluation Criteria for Toxldty

          (1)  Complete toxicity

          When there are no revertants observed on the plate(s) treated
          with the test compound, the test compound is defined as toxic
          to all or any of the indicator strains at that (those) particular
          dose(s).

          (2)  Slight toxicity

          When there are fifty or less percent revertants on the plate(s)
          treated with the test compound as compared to the solvent control
          plate(s), the test compound is defined as slightly toxic to
          all or any of the indicator strains at that (those) particular
          dose(s).

     F.   Relation Between Mutagenicity and Carcinogenicity

     It must be emphasized that the Ames Salmonella/Microsome Plate Assay
     is not a definitive test for chemical carcinogens.   It is recognized,
     howerver, that correlative and functional relations have been demon-
     strated between these two endpoints.   The results of comparative
     tests on 300 chemicals by McCann et.  aJL1 show an extremely good
     correlation between results of microbial mutagenesis tests and j_n
     vivo rodent carcinogenesis assays.

     All evaluations and interpretation of the data to be presented in
     the final report will be based only on the demonstration, or lack,
     of mutagenic activity.
                                  166

-------
                                REFERENCES


1.   McCann,J., Choi, E., Yamasaki,  E.  and Ames, B.N.:  Detection of
     carcinogens as mutagens  in the  Salmonella/microsome test:  Assay
     of 300 chemicals.   Proc. Nat. Acad. Sci. USA, 72:5135-5139, 1975.

2.   Ames, B.N., Gurney, E.G.,  Miller,  J.A. and Bartsch, H.:  Carcinogens
     as frameshift mutagens:  Metabolites and derivatives of 2-acetyl-
     aminofluorene and other  aromatic amine carcinogens.  Proc. Nat. Acad.
     Sci. USA,  69:3128-3132,  1972.

3.   Ames, B.N, Lee, F.D. and Durston,  W.E.:  An improved bacterial
     test system for the detection and  classification of mutagens and
     carcinogens.  Proc. Nat. Acad.  Sci. USA, 70:782-786, 1973.

4.   Ames, B.N, Durston, W.E.,  Yamasaki, E. and Lee, F.D.:  Carcinogens
     are mutagens:  A simple  test system combining liver homogenates
     for activation and  bacteria for detection.  Proc. Nat. Acad. Sci.
     USA, 70:2281-2285,  1973.

5.   McCann, J., Springarn, N.E., Kobori, J. and Ames, B.N.:  Detection of
     carcinogens as mutagens:   Bacterial tester strains with R factor
     plasmids.  Proc. Nat. Acad. Sci. USA, 72:979-983, 1975.

6.   Ames, B.N., McCann, J. and Yamasaki, E.:  Methods for detecting
     carcinogens and mutagens with the  Salmonel1a/mammalian-microsome
     mutagenicity test.  Mutation Res., 31:347-364, 1975.

7.   Vogel, H.J. and Bonner,  D.M.:   Acetylornithinase of E. coli; Partial
     purification and some properties.  J. Biol. Chem., 218:97-106, 1956.
                                 167

-------
                      Q.A. Inspection Statement
                   (reference 21 CFR 58.35(b)(7))
PROJECT   3-C*!??                          LBI Assay No.
TYPE of STUDY       ft Mrs  7/^-rr  Tfsr~
         This final study report was reviewed by the LBI Quality
Assurance Unit on     /£ / ^ / fo         A report of findings was
submitted to the Study Director and to Management on    /3~/ s
         The short-term nature of this study precluded inspection while
it was in process.  The Quality Assurance Unit inspects -an in-process
study of this type approximately once per month to assure that no
significant problems exist that are likely to affect the integrity of
this type of study.
                                      Auditor, Quality Assurance Unit
                                      168

-------
                         GENETICS ASSAY NO.   5394
                             LBI SAFETY NO.   6378
     TOXIC EVALUATION OF
         SAMPLE #1452
(WATER TANK DRAIN,  END OF TEST)

            IN  THE
    RODENT QUANTAL  TOXICITY
             ASSAY
         FINAL REPORT
         SUBMITTED  TO:

      ACUREX CORPORATION
       485 CLYDE  AVENUE
   MOUNTAIN VIEW, CA 94042
         SUBMITTED BY:

    LITTON BIONETICS,  INC.
     5516 NICHOLSON LANE
 KENSINGTON,  MARYLAND   20795
     LBI  PROJECT NO.  22064


 REPORT DATE:   DECEMBER,  1980
        169

-------
                            PREFACE
This report contains a summary of the data compiled during the
evaluation of the test compound.   The report is organized to
present the results in a concise  and easily interpretable manner.
The first part contains items I - VIII.   Items I - III provide
sponsor and compound identification information and identify the
type of assay.  The assay was conducted  and evaluated according to
procedures recommended in IERL-RTP Procedures Manual:  Level 1
Environmental Assessment Biological Tests (Litton Bionetics, Inc.,
Kensington, MD, September, 1980,  in press).  Item IV provides the
initiation and completion dates for the  study, and Item V provides
identification of supervisory personnel.  Item VI identifies the
tables and figures containing the data used by the study director
in interpreting the test results.  The interpretation itself is in
Item VII.  Item VIII provides the conclusion and evaluation.

The second part of the report, entitled  Assay Design, describes the
materials and procedures employed in conducting the assay.   This
part of the report also contains  evaluation criteria used by the
study director, and any appendices.  The evaluation criteria are
included to acquaint the sponsor  with the methods used to develop
and analyze the test results.

All test and control results presented in this report are supported
by fully documented raw data which are permanently maintained in
the files of the Department of Genetics  and Cell Biology or in the
archives of Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington
Maryland, 20795.

Copies of raw data will be supplied to the sponsor upon request.
                               170

-------
   I.   SPONSOR:   Acurex Corporation

  II.   MATERIAL  (TEST COMPOUND):   GENETICS ASSAY NO.:  5394

       A.   Identification:  Sample #1452 (Man Test 5/21/80,  #5  Gal, water tank
       D   n +  B   •  j   «              drain, end of  test)
       B.   Date  Received:  October 6, 1980

       C.   Physical  Description:   Clear, colorless liquid

 III.   TYPE OF ASSAY:  Rodent Quantal Toxicity Assay

  IV.   STUDY DATES:

       A.   Initiation:   October  22, 1980

       B.   Completion:   November 24, 1980

   V.   SUPERVISORY PERSONNEL:

       A.   Study Director:  David J. Brusick, Ph.D.

       B.   Laboratory Supervisor:  Joan McGowan

  VI.   RESULTS:

       The data  are  presented in  Table 1 on pages 3 and  4.

VII.    INTERPRETATION OF RESULTS:

       The test  material, Sample  No. 1452, was concentrated  10-fold (as
       described under Assay Design) prior to administration by oral gavage
       to  10 male and 10 female weanling CD-I mice.  A single average dose
       of  5 ml/kg was given, which corresponded to 0.08  ml for  the males
       and 0.07  ml for the females.  The animals were observed  for 14 days
       for toxic signs or lethality.  Survivors were weighed and necropsied
       on  Day 14 after dosing.

       All twenty animals survived the exposure with no  evidence of any
       compound-related toxic signs.  Both the male and  female  mice showed
       good weight gains, and no  observations were obtained  at  necropsy
       that indicated compound-related lesions (Table 1).  The  test material
       was therefore considered to have no detectable toxicity  in this assay,
       and according to the Evaluation Criteria, an LD50 determination was
       unnecessary.
                                    171

-------
VIII.  CONCLUSIONS:
       The test material,  Sample No. 1452, concentrated 10-fold, had no
       detectable toxicity to weanling CD-I mice in the Rodent Quantal
       Toxicity Assay.   The applied dose was 5 ml/kg.
                                         Submitted  by:

                                         Study Director
                                     ,               £    ^)       U/3/lo
                                    JLf '. David J.  Brusick, Wi.D.     Date
                                         Di rector
                                         Department of Genetics
                                         and Cell  Biology
                                      172

-------
                                  TABLE 1

                 QUANTAL TOXICITY DATA WITH WEANLING MICE
              Quanta!  Toxicity:  Weanling mice
              Sponsor:  Acurex Corporation
              Test Article:  Sample No.  1452
              Vehicle:  Not Applicable
              Study Dates:  11/10/80 to  11/24/80
              Animal Room No.:  4S
              Animals:  Charles River CD-I mice,  P.O.  100760
              Dose:  5 ml/kg administered P.O.

Initial
Height
Animal No. gm
Final Visible
Weight Toxic
gm Signs3


Gross Necropsy Findings
Males
  1101
  1102
  1103
  1104
  1105
  1106
  1107
  1108
  1109
  1110
           24.2       NTSb      Small mucoid lump in bladder
           25.5       NTS      Both eyes opaque
           22.1       NTS      Small mucoid lump in bladder
           21.7       NTS      NSLC
           23.2       NTS      NSL
           22.2       NTS      Both eyes opaque
           23.7       NTS      NSL
           23.7       NTS      NSL
           23.8       NTS      -Both eyes opaque
                               •Mesenteric lymph nodes
                                slightly enlarged
                               •Left kidney slightly pale
16.6       25.2       NTS       Both eyes opaque
16.1
16.7
14.2
14.4
15.6
13.
16.
13.0
14.8
.5
.2
  Mean Body Weight:
           Initial  15.1 ± 1.3 gm (Standard Deviation)
             Final  23.5 ± 1.3 gm (Standard Deviation)
a Animals observed for 14 days.

b NTS = No Toxic Signs.

c NSL = No Significant Lesions.
                                     173

-------
                           TABLE 1  (continued)

                QUANTAL TOXICITY DATA WITH WEANLING MICE

Animal No.
Females
1111
1112
1113

1114
1115
1116

1117
1118
1119

1120
Initial
Weight
gm

14.1
14.4
14.3

14.5
12.5
14.0

13.3
12.1
13.5

11.9
Final
Weight
gm

19.8
20.0
20.4

20.2
19.0
21.2

18.7
19.9
19.7

16.7
Visible
Toxi c
Signs3

NTSb
NTS
Possible nasal
irritation
NTS
NTS
NTS

NTS
NTS
NTS

NTS
Gross Necropsy

NSLC
NSL
Uterine horns
fluid
NSL
NSL
Uterine horns
fluid
NSL .
NSL
Left kidney si
smaller than
NSL
Findings



enlarged/



enlarged/



ightly
right

Mean Body Weight:
               Initial
                 Final
13.5 ± 1.0 gm (Standard Deviation)
19.6 ± 1.2 gm (Standard Deviation)
aAnimals observed for 14 days.

bNTS - No Toxic Signs.

CNSL = No Significant Lesions.
                                    174

-------
                              ASSAY  DESIGN

                             PJJANTAL IOXICITY ASSAY
SllLa+-ay S0nf0rm?rJ^°?fn?f the b1oassay tests in the US Environmental
Protection Agency  (EPA)  IERL-RTP Procedures Manual :  Level 1 Environ-
mental Assessment  Biological Tests  (September. TQan)   »o»0i i Mna
obtain preliminary information on the harmful biological effects of
chemicals found in industrial feed  and waste streams.


1.  OBJECTIVE

    The objective  of this assay was to evaluate the acute toxicity of a
    test material  when administered by oral gavage to male and female
    weanling mice.

    The assay consisted  of recording any lethality and toxic signs that
    occurred over  a 14-day period following a single treatment and then
    collecting necropsy  information on animals that died or were killed
    at the end of  the observation period.


2.  TEST MATERIAL

    A test material described as Sample #1452 (Man Test 5/21/80, #5 Gal,
    Water Tank Drain, End of Test) was received as a clear, aqueous
    liquid containing no obvious particulate material.   The sample was
    stored at 4°C  in its original one-gallon amber glass container.

    An aliquot of 600 ml of test material was concentrated 10-fold to
    60 ml by lyophilization prior to testing.  A total  volume of 1.5 ml
    of the concentrate was then utilized for oral  dosing of a group of
    20 weanling mice.  The concentrate was stored  at 4°C in an amber
    glass bottle.
3.  EXPERIMENTAL DESIGN

    Nine nursing female Charles River CD-I  mice with six  pups  each  (three
    male and three female) were obtained from Charles River  Breeding
    Laboratories, Inc., Wilmington, Massachusetts on November  5,  1980.
    The birth date of the pups was October  19, 1980.  The animals were
    quarantined for 5 days upon receipt. The litters were individually
    housed on Absorb-Dri bedding in polycarbonate cages and  were  cared
    for according to Litton Bionetics, Inc.,  Department of Genetics and
    LAMS Standard Operating Procedures.   Purina certified laboratory
    chow and water (pH 2.5) were provided ad  libitum. The pups were
                                     175

-------
3.  EXPERIMENTAL DESIGN (continued)


    maintained with mothers until  weaned.   The animals were identified by
    eartags and cage cards and were released for study on November 10,
    1980.

    Prior to initiation of dosing, ten male and ten female weanling mice
    were individually weighed; the volume  of the test material  to be
    administered was based on the  mean weight values for each sex.  The
    test material (10-fold concentrate of  Acurex Sample #1452)  was ad-
    ministered once at 5 ml/kg on  November 10, 1980.  The weanling mice
    were 23 days old.  The animals were observed at the time of dosing,
    2 hours later, and daily thereafter.  On day 14, the surviving animals
    were fasted overnight.  They were subjected to gross necropsy the
    next day.
4.  ASSAY EVALUATION CRITERIA

    Immediately following administration of the test material  and at
    frequent intervals during the first day, observations  of the frequency
    and severity of all toxic signs or pharmacological  effects (as listed
    in Table A) will be recorded.  Particular attention will be paid to
    the time of onset and disappearance of the signs.  Daily observations
    will be made and recorded on all animals through a  14-day period.  At
    termination of the observation period, all surviving animals will  be
    weighed, killed, and then gross necropsies performed.   Necropsies  will
    also be performed on all  animals that die during the course of this
    study.

    If no mortality occurs in the quantal study, no further studies will
    be performed with the test substance and the LDso should be reported
    as greater than 5 ml/kg or 5 g/kg.  The test material  is ranked as
    having nondetectable toxicity (ND) at the maximum applicable dose (MAD),
    Effluent samples which produce harmful effects in vivo and do not
    result in deaths will be noted in the results summary.  Such observa-
    tions are difficult to quantitate but provide insight  into the sub-
    lethal effects of a sample on rodents.  Further investigations may be
    recommended from observations of nonlethal toxic effects.

    If a single animal in the quantal study dies in the 14-day observation
    period, a quantitative study will be performed.  An LDso will be
    calculated by the method of Litchfield and Wilcoxin.  If the data are
    not suitable for calculation of a precise LDso, i.e.,  total mortality
    occurs for the lowest dose, an estimate of the LDso could be made or
    the LDso could be expressed as 0.05 ml/kg or 0.05 g/kg or less.  Oc-
    casionally, it may be necessary to use a different  series of dosages
    in a repeat study to accurately locate the LDso. The  calculated LDso
    value is used to rank the toxicity of the test material according to
    the dose ranges presented in Table B.
                                    176

-------
             TABLE A.    DEFINITION OF PHARMACOLOGICAL TOXIC SIGNS
  Organ System
   Observation and
     Examination
    Common Signs of Toxicity
CNS and
somatomotor
Autonomi c
nervous system

Respiratory
 Cardiovascular
 Gastrointestinal



 Skin and fur

 Mucous membranes

 Eye

 Others
Behavior
                     Movements
Reactivity to various
stimuli
Cerebral and spinal
reflexes
Muscle tone
Pupil size

Secretion
Nostrils
Character and rate
of  breathing

Palpation of cardiac
region

Events
abdominal shape
feces consistency
and color   ,
Vulva,  mammary
glands
Penis
 Perianal  region
Color,  turgor,
 integrity
 Conjunctiva,  mouth

 Eyeball
 Transparency
 Rectal  or paw skin
 Injection site
 General Condition
Change in attitude to observer,
unusual vocalization, restless-
ness, sedation
Twitch, tremor, ataxia, cata-
tonia, paralysis, convulsion,
forced movements
Irritability, passivity,
anaesthesis, hyperaesthesis
Sluggishness, absence

Rigidity, flaccidity
Myosis, mydriasis

Salivation, lacrimation
Discharge
Bradypnoea, dyspnoea, Cheyne-
Stokes breathing, Kussmaul
breathing
Thrill, bradycardia, arrhy-
thmia, stronger  or weaker
beat
Diarrhea, constipation,
Flatulence, contraction
Unformed, black  or clay colored

Swelling
 Prolapse
 Soiled
 Reddening
 eruptions
           flaccid skinfold,
           pi 1oerection
Discharge, congestion,
hemorrhage cyanosis, jaundice
Exophthalmus, nystagmus
Opacities
Subnormal, increased temperature
Swelling
Abnormal posture, emaciation
                                        177

-------
4.  ASSAY EVALUATION CRITERIA (continued)


    Observations are also made and recorded daily on all  animals through
    the 14-day period.  As in the quantal phase, no attempt is made to
    quantitate or rank the observations.   The average animal  body weight
    of each group is determined initially and at the termination of the
    experiment.  The average weights and  the weights as fractions of the
    control are reported for each dose level.  Necropsy observations are
    recorded and reported.


        TABLE B.  ACUTE IN VIVO RODENT ASSAY EVALUATION CRITERIA
                             Solids                     Liquids
      Toxicity3           (LDso in g/kg)             (LDso in ml/kg)
High
Moderate
Low
Not Detectable
<0.05
0.05 to 0.5
0.5 to 5
>5
<0.05
0.05 to 0.
0.5 to 5
>5

5



    Evaluation criteria formulated by Litton Bionetics, Inc.  for IERL-
     RTP Procedures Manual:  Level  1  Environmental  Assessment  Biological
     Tests.
5.  RECORDS TO BE MAINTAINED

    All raw data, protocols, protocol  modifications,  test article weight
    and dispensation records and correspondence between LBI and the Sponsor
    are being maintained in a central  file within the Department of
    Genetics and Cell Biology.  These  records will  be filed under depart-
    mental assay number and held up to 2 years following submission of the
    final report to the Sponsor.  After 2 years they  will be transferred
    to the LBI archives for permanent  storage.
                                     178

-------
                      Q.A. Inspection Statement
                   (reference 21 CFR 58.35(b)(7))
PROJECT    1G                                LBi Assay No.
TYPE of STUDY     ?0e/eW  Quwr*t.TaK,c;
         This final study report was reviewed by the LBI Quality
Assurance Unit on     )£ //'/So _ .  A report of findings was
submitted to the Study Director and to  Management on    t
         The short-term nature' of this study precluded inspection while
it was in process.  The Quality Assurance Unit inspects .an in-process
study of this type approximately once per month to assure that no
significant problems exist that are likely to affect the integrity of
this type of study.
                                      Auditor, Quality Assurance Unit
                                      179

-------
    SECTION 6



SAM IA WORK SHEETS
      180

-------
  I
1. SOURCE/CONTROL OPTION:
                                                                                               tgVR. t
                                                                                               LEVEL 2
». EFFLUENT STREAM
  CODE NO.
  NAME
                                                             3. TOTAL MASS RATE OF DISCHARGE
4. COMPLETE THE FOLLOWING TABLE FOR THE EFFLUENT STREAM OF LINE 2
                         C
     n
                is
SKOESOM
CATEGOMV
NCENTRATION
COkOOiCAL
OM

VERITY
HEALTH
(C.'O)
DIS
SE
-
DISCHARGE
SEVERITY
ECOLOGICAL
iC/f)
                                                                                                     WCMXTED IMOMMC SC«IMTT
    UMTS
               01
           2_
                                           p//>a«o

                                                                    t*fio
                                                                           *
              CrtA
l-oito3-
                                           09.000
                                                                                                     I'M***1
 ^O-

                                                                                                     1-21*10
               27
                                                                                                      /•i
I* MORE SPACE IS MEMO USE A CONTINUA1ION SHCEf I
 S TOTAL DISCHARGE!
   HEALTH BASED ( r col H) &a
   ECOLOGICAL BASED ( I col 11 So    .     .  	
   (ENTER HERE AND AT LINE 8 OF THE SUMMARY SHEET)
                                         6 TOTAL WEIGHTED DISCHARGE SEVERITY
                                           HEALTH BASED | £ col LI 6a	
                                           ECOLOGICAL BASED (I col M| 6b	
                                           (ENTER HERE AND AT LINE 8 OF THE SUMMARY SHEET)
-•/»
_•/•

-------
          OMTMUMWH
                          torn vmxtMCcr
          1. SOUMCf/CONTHOt OPTION
           COM NO
                                                                                                   LCVCL I.
                                                                                                   LfVCl.2.
                         NAME
              f
                        it
WCOUM
e«TiGOTf
NCIMTMIlOM


                                                                                                    •MntU


                                                   It
                                                                      f
                       £29
                       3ft
                        tl-OP/O*
                                                                                                    a./
                                       J-SL^tO*
                                                             _
                                                         -3*10
                      31
00
tV)
                      33
                          .     3
                         6 -o* to

                                                                                                    6.2*10
                      3*

                                                                                      3-3*10*
                      35"

                                         0+10*
                                                                            -a

                       ar
         8 flu-t
urn
         31
So+lo

-------
         COM NO.
                           NAME
                                                                                                JL.
            fj


            S3

            ii
  „;

   r-
+
  i
  I
                                                                                       »
                                                            1 *


           42.
            flCSflZ
tt-O+IO*
                                    OBiae


                                            fe&azg
                                                                       "'

                                                                                           * *1**lo
                    {&.
                       o
                  :fta*/c>
          (£P£
                           /«/K/0

                           ^•0"/o'
         fc^^00
00
co
         i£tt£.
                           29
                           '•****'
         MSL.
'^fUof^u
                           2-2
         »&*>0
?***>
                                                                       '*
         flMriMt,*/*/
                                    ssasoii

                                   jLssta£..
         Soy.
                                                                                    9.8*10*

-------
          *+** HMI »O»MMMT
1. SOUMCbCOMf MM. OPTION
                                                                                      »f L I.
t IMLOCMT.TfttAU
COM NO
   i!
'

i!
                                                     i"
                                                                                *
   iwn
                           «e
                                                                                      1.3*10
                                                                                            '

                  }•**•* to
       2-0* /
-------

           1 WXJNCMONf MM. OPTION
                               t 0 # /

                        /*t 0 #  HaT> » >cK.
           COM NO.




                                                                  if
                                                 n-
           ItOfl
*
         AOX /P
                               /. 5*

                       7*+

                       79
               3-o
                                        3 -0*10
                                                                      3-0
00
01
 ?
f+y/o
                                    3
lo+to
                       €1
                        Suttee
                       83.
                                                                      /•€>»./£?


-------
      E/CONTROL OPTION:               *

                 tuMAce  /y? A-Al-
                                                                                                     ,.,.-. ,
                                                                                                     LEVEL 1
                                                                                                     LEVEL f
          1 IFFLUtNt STMAM
            OOOCNO.
                                                                    3 TOTAL MASS RATE OF DISCHARGE
                                                                       O» -  -   -
          4. COMPLETE TMf fOUOMHNO TABLE FOR THE EFFLUENT STREAM Of LINE 9


               H

                                 o
                                 SE

                                                                                               MMHIU
VIWT
MEAkT
UV
f CQl
                              tt.ot.icr1
                                                                          ^•0*10

                                                                                                                  -8
                                               /g8»/o
oo
CTl


                                         a-two^
                                                       3£££g.

                              3-0K./01
                               St-o
                            ff-Jt-x/o*
                                    / 1*10
                                                                                                                 "*
                       M-t,
                   "70X/01

         l*MOMtMCfMM MOUM ACOMfMUA1IOMSI«It|
          S TOTAL DISCHARGE
            HEALTH BASED ( f col
            ECOLOGICAL BASED 11 col I) Sto
                             /•a^/c*Z_
(ENTER HERE AND AT LINE • OF THE SUMMARV SHEET)
                                                         • TOTAL WEIGHTED DISCHARGE SEVERITY
                                                           HEALTH BASED 11 col LI ft*	
ECOLOGICAL BASED ( £ col M| Cfc	& • «
(ENTER HERE AND AT LINE • Of THE SUMMARV SHEET|

-------
             COM NO.
                 H
                 si
                 u5
                 (a
            "

H
i?
if
                                                                                     <
                                                                                     8
                                 ^
          ^
                                          7-Sv.lct*

                                J-o*. to
                                   SJBiltD
                                                                            3
oo
                                                    SMUoo
                         STOOD
          /Vx/o5"
                   STtUoO
                          ss
Sox/c
                          LSL

                                3 ox /o

-------
                                                                                                      *~^* J   *f*
                                                                                                        *L
            ft If riUfNt SfMAM
            COM NO
                                         W*ff*.
                        -o  Sr/tfM/t^
                                 .£-1
               |f

               !i
               ts



                      I
                  I"
                            .

                         1"
III
                                         4#
                        LS
          2-Sx'c?
b&loco

                                                          -
                                                     -7,5*10
                         tfi
•70*10*
                        3-8
                                                I/

                                                  (A/toot)
                                                                                                                 £•0*10
                                   lO
                    TbAeeu
00
oo
                        -rl
  9*. so

                                   I'1?
                        -73.
          l-Svtct*
                                 Jt.-ot.io
                                                                                              I/
                                                  TWlCC)
                                                        2-SxtO
                                                                                                              -1
                                                                  76*00
                                        S-Cfa /O
                                           /CO-
                        79
                        €1
       j£a	



       11*10*
                                    Tfflooo
                                                      Lo

-------
           COM NO.

              IE




                                  It
                                                                       K-

              rtHUMtt


                 $w°
                                                                                                £.6*10
                                                                                                       *'
oo
vo
          ££/*Ht#r
&0_

8*4000
                                                                          '

-------
                                TECHNICAL REPORT DATA
                         (Pleau nod Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/7-82-038b
                           2.
                                                      3. RECIPIENT'S ACCESSION-NO.
4. TITLE AND SUBTITLE
 Environmental Assessment of a Low-emission oil-
  fired Residential Hot Water Condensing Heating
  System; Volume II Data Supplement	
                                                      5. REPORT DATE
                                                      May 1982
                                                      6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)

 C. Castaldini
                                                      B. PERFORMING ORGANIZATION REPORT NO
                                                      10. PROGRAM ELEMENT NO.
9. PERFORMING OROANIZATION NAME AND ADDRESS
 Acurex Corporation
 485 Clyde Avenue
 Mountain View, California 94042
                                                      11. CONTRACT/GRANT NO.
                                                      68-02-3188
 12. SPONSORING AGENCY NAME AND ADDRESS
 EPA, Office of Research and Development
 Industrial Environmental Research Laboratory
 Research Triangle Park, NC 27711
                                                      13. TYPE OF REPORT AND PERIOD COVERED
                                                      Final: 7/80-2/81	
                                                      14. SPONSORING AGENCY CODE
                                                       EPA/600/13
 is SUPPLEMENTARY NOTES !ERL.RTp project officer is Robert E. Hall, Mail Drop 65, 919/
 541-2477. Volume I. gives technical results.
 i6. ABSTRACT The report gives results of a test program measuring air and water emis-
 sions from a high-efficiency hot-water residential heating system of European
 design, utilizing a condensing flue gas system and a low emission burner. Criteria
 and noncriteria emissions,  including trace elements and organic species in both flue
 gas and condensate waste water streams, were measured. NO (as NO2), CO,  total
 UHC (as propane), and total particulate emissions measured about 37, 12, 1. 5, and
 2.7 ng/J heat input,  respectively. Absorption of sulfates and nitrates in the waste
 water resulted in a constant pH of 3.0. Total organic emissions in the flue gas mea-
 sured 3.5 mg/dscm; they were below the detectable limit in the waste water. Several
 inorganic trace elements, including  chromium,  copper, iron, and nickel, in the
 waste water were attributed to leaching of heat transfer metal surfaces by the warm
 acidic water. Bioassays were also performed to evaluate the potential health hazard
 of the streams. Results indicate nondetectable to moderate toxicity and mutagenicity.
17.
                             KEY WORDS AND DOCUMENT ANALYSIS
                DESCRIPTORS
                                         b.lDENTIFIERS/OPEN ENDED TERMS
                                                                  c. COSATI Field/Group
 Pollution           Waste Water
 Assessments       Condensates
 Hot Water Heating  Measurement
 Residential Buildings
 Fuel Oil
 Flue Gases
                                          Pollution Control
                                          Stationary Sources
                                          Environmental Assess-
                                           ment
13B
14B
13A
13M
21D
21B
07D
13. DISTRIBUTION STATEMENT

 Release to Public
                                         19. SECURITY CLASS (This Report)
                                          Unclassified
21. NO. OF PAGES
    193
                                         20. SECURITY CLASS (Thitotge)
                                          Unclassified
22. PRICE
EPA Form 2220-1 (t-7J)
                                        190

-------