RATIONAL BmLI:HE DIWIS'J:m
Bio-Assay Inves t iga t ions
National Anil ine Division
Al l i ed Chemical and Dye Corporation
Buffalo, New York
Prepared by
Croswell Henderson, Aquatic B io log i s t
San i t a ry Engineering Center, USPHS
Cincinnat i , Ohio
and
H. A. Anderson, Pub l i c Health Engineer
IJC F i e l d Unit, UPHS
Buffalo, Mew York
In t roduct ion
Purpose
I n January 1956, f i e l d s t u d i e s were i n i t i a t e d o n = cooperat ive b a s i s
t o determine the possible e f f e c t s of the wastes.from major Buffalo River
i n d u s t r i e s on t h e e a s t e r n end of Lake E r i e and t h e Niagara River. Buffalo
River i n d u s t r i e s cooperating i n t h i s p r o j e c t were the National k n i l i n e
Division-Allied Chemical and Dye Corporation, Donner-Hanna Coke Corporation,
and the Socony k b i l O i l Company.
This r epor t covers an i n v e s t i g a t i o n of the t o x i c i t y t o f i s h of i n t a k e
waters and major e f f l u e n t s from the National Ani l ine Division-Allied Chemical
and Dye Corporation. Chemical and dye was,tes con ta in some chemical compounds
which a r e known t o be t o x i c t o a q u a t i c l i f e i n 'low concent ra t ions . Some of
these chemicals when mixed wi th o r under t h e in f luence of o the r non-toxic
components oY'the e fy luen t o r rece iv ing water may e x e r t a n e n t i r e l y d i f f e r e n t
toxic i - ty from t h a t oT t h e pure compounds. Bio-assays were m d e t o eva lua te
d i r e c t l y t h e t o x i c i t y of t hese chemically complex wastes .
Personnel P a r t i c i p a t i n g
National Anil@e Uivis ion
Dr. it. L. Hess, Coordinator of P o l l u t i o n Fiesearch
C . J. Carney, Chemist,
J, A. Gouck, Chemist
Pub l i c Health Serv ice
Hayse H o Black, I n d u s t r i a l Wastes Consultant
Croswell Henderson, Aquatic B io log i s t
D r . C . M. Tarzwell, Chief, Aquatic Biology Unit
H o A. Anderson, Pub l i c Heal th Engineer
M o W. Ruszaj, Chemist
Organizat ion - of Study
National Ani l ine Divis ion personnel c o l l e c t e d 2h hour composite samples
of t h e i r i n t a k e water and e f f l u e n t s and determined t h e volume of e f f l u e n t s
discharged. !The Pub l i c Heal th ~e l -v ide conducted bio-assays i n l a b o r a t o r y
space provided by National Ani l ine a& made phenol and cyanide de terminat ions
i n t h e I J C Unit l dbora to ryo Phenol and cyani'de de terminat ions were a l s o made
on some e f f l u e n t samples-by t h e National Anil ine P o l l u t i o n Research Laboratoryo
r 3 -
General Information
Production Operations
National Aniline Division is one of the largest producers of dyes, dye
intermediates, and synthetic organic chemicals, including surface active agents,
t e x t i l e assistants, p las t ic intermediates, and pharmaceuticals i n the United
States, Batch type operations are used in the production of many of the numerous
products of the Buffalo plant. The products being manufactured a t any one time
vary greatly according to market demand and stock on hand,
Water Supply #
Buffalo city water is used for drinking, sanitary purposes, and as process
water, The major portion of the plant water supply is used for process cooling
and is obtained from the Buffalo River through a shore intake located about 100
fee t upstream from the National Aniline Plant nAtt Sewer ou t fa l l and downstream
from plant W n , and flEn sewers. During periods of low r iver flow, there is some
recycling of the plant effluents,
Process Wastes
Wastes are discharged into the Buffalo River through three separate outfal le
designated as plants tlAtt, nBB&Bfl, and nEn sewers, A l l of these sewer out fa l l s are
on the North bank of the Buffalo River. Plant nAu outfal l i s located 400 fee t
downstream from South Park Bridge, plant nBfbCn outfal l immediately south of the
bridge, and plant ou t fa l l about 1000 fee t upstream from the bridge.
Each of the above sewers receives wastes from numerous and varied productf on . processes. The wastes i n each may be expected to vary significantly a t times due
to changes in 'quantity o r type of products being manufactured..
Waste Control Mearmre s
1, l o r the past 20 years the company has supported and continues t o support
a program of research on the effect s of and methods f o r t r ea t ing i t s wastes and i t s f u l l scale application t o reducing e x i s t ing pollution and minimizing or avoid- ing pol lut ion by new processes.
2, BOP t h i s purpose 811 especially equipped laboratory hae been provided and manned with special ly t ra ined personnel f o r the en t i re period.
3, Duri ng recent yews a p i lo t plant has been provided and operated f o r in- vest igat ing the chemical and biochemical treatment of objectionable wastes which might resu l t from expansions of production i n the future.
b0 This program is integrated into the en t i re organization.
5 New Processes, equipment i n s t a l l a t i ons and replacements ape not operated u n t i 1 approved fo r wastes control by the Pollution Research Unit.
I
6. Two un i t s for neutral izat ion of waste acids with lime have been constructed and a re operated when necessary to maintain a pH of at least 6 i n the Buffalo River.
7, Some wastes are segregated and discharged to the Buffalo Sewage System, with wri t ten approval by the Buffalo Sewer Authority, for which fees a re paid.
8. Wastes containing arsenic are placed i n containers and disposed of by bur ia l 100 miles at sea.
9. . Certain wastes, including sulf ides and thioeulfates, ape destroyed by cherm- i c a l treatment i n especially designed equipment,
10. Some o i ly wastes are separated and used a s f ie l .
11, Some o i l s , tws, and various other kinds of combustible wastes w e separ- ated and burned i n an especially designed incinerator.
12. Non-combust i b l e so l ids and sludges, includi ng metal l i e compounds, a re sep- a ~ a t e d and deposfted i n dumps or sludge ponds fo r eventual sa le or dfsposal i n a manner t o avoid polluffon,
13. Aluminum chloride wastes are sent t o the Buffalo Sewage Treatment Plant where they are used to advantage a s a sludge conditioner,
14, Other wastes are recovered, reff ned and used i n plant production processeso
. . . Survey Methods
Plow Measurement8
Waste flow i n the sewers was determined by Nat ional Aniline personnel. Flow
i n the Plant Sewer was determined by a water level recorder which meamred the
height of the l iqu id i n a sewer. This deviee had previously been calibrated by
P i to t tube measurement 8,
Flows i n p lan ts "8" and nBBd3n Sewers were estimated from the volume of intake
water. City intake water was measured by pos i t ive displacement meters and r iver
water intake was measured by o r i f i c e meters.
Following a re the measured da i ly flows during a one week period. The average
valuea were used i n a l l bio-assay compatati one:
1000 gal ./day
Date Plant MA# Plants MBKGM Plant nEn
1-19 11,349 7, 231 1,753
1-20 11,265 7 9 341 1,680
1-21 11,225 6 9 113 1 s 770
1-24 11,151 6,529 2,090
1-25 11,386 6,898 l o 918
Average mgd 11.30 . 6-82 1.84
c f s -
S a n p l i q
Twenty-four hour composite effluent samples were col lected by means of =to-
matic samplers. Samples of Plant nAVewer were collected a t the ou t fa l l . Plant
nB&Cn samples were collected through a r f se r located 15 f ee t upstream from the
sewer outfal l . Plant flEw samples were collected from %he influent t o the waste
treatment pf lo t plant. This inf luent i s pumped from a point i n the sewer approxi-
mately l O O O 1 above the sewer mouth.
Chemical Methods
A 1 1 chemical determenat ions were made accordi ng t o procedure s i n '%tandwd
Methods for the Examination of Water, Sewage, and Industr ia l Wastes#, 10th edition,
published by the American Public Health Association,
Bio-assay Methods
Bio-assays were made, e s s e n t i a l l y , by t h e method recommended by the
t o x i c i t y subcommittee o'f t h e Federat ion of Sewage and I n d u s t r i a l Wastes
Associations (sewage and I n d u s t r i a l Wastes,' Vol. -23, No. 11, 13809 Nov. 1951) . This method c o n s i s t s of preparing various, concentrat ions of e f f l u e n t i n a
se l ec ted d i l u t i o n water, adding the t e s t f i s h and observing t h e i r r e a c t i o n s
over a d e f i n i t e time period. A logar i thmic s e r i e s of numbers i s genera l ly
most convenient f o r preparing d i f f e r e n t t e s t concentrat ions.
A s t h e e f f l u e n t s were of unknown t o x i c i t y , explora tory o r small s c a l e
t e s t s were made t o de termine . the approximate tox ic range. Test s o l u t i o n s (
were prepared over. a wide range of concent ra t ion (e .go 100, 1, and 0.1 percent
e f f l u e n t ) . Two f i s h were added t o 2 l i t e r s of each concentrat ion i n 6-1/2 inch
diameter, 1-ga l lon wide mouth g l a s s b o t t l e s . Observations f o r r e l a t i v e l y s h o r t
time periods f ndicated t e s t concentrat tons necessary f o r t h e f u l l s c a l e experiments,
I n t h e f u l l s c a l e t e s t s , 10 f i s h were gene ra l ly used f o r each t e s t
concentrat ion. Five f i s h were added t o 10 l i t e r d u p l i c a t e samples i n 10 inch
diameter, $gallon widemout)l g l a s s b o t t l e s . ?he intermediate concent ra t ions
t e s t e d were dependent upon information obtained from the explora tory t e s t s .
For example, if f i s h were k i l l e d i n concent ra t ions above 1 0 percent and not
a f f e c t e d i n concentrat ions of 1 percent, in termedia tes were s e t 'up within t h i s
range (e.g., 10, 5.6, 3.2, 1.8, and 1.0 aercent concentrat ion of e f f l u e n t ) .
The d i l u t i o n water used was raw Lake E r i e water obtained a t the I
Buffalo, N. Y., water p l an t . This water was hauled i n t o t h e labora tory ,
allowed t o come t o room temperature, and ae ra ted vigorously f o r a t l e a s t one
h o w t o b r ing it t o equil ibr ium with atmospheric gases. C h a r a c t e r i s t i c s of t h i s
water a t time of use were a s followss Dissolved oxygen 7.6-8.2 ppm; pH 8.0-8.2;
Tota l a l k a l i n i t y ( C ~ C O ) '94-100 ppm; To ta l a c i d i t y ( c ~ c o ~ ) 0-1 ppm; Versenate 3
hardness (CaCo3) 120-135 ppm.
The t e s t f i s h used were fa thead minnows ( ~ i m e p h a l e s ~ r o m e l a s ) of a
f a i r l y uniform s i z e , ranging in length from 2 t o 2-1/2 inches and i n
weight from 1 t o 1-1 f2 grams. These f i s h were obtained i n uniform l o t s
from the Newtown, Ohio, 'Fish Hatchery and accl imated t o l abora to ry condit ions.
This spec ie s is of in te rmedia te to l e rance to chemicals comparable t o bass,
sunf ish , perch, and o the r warm water spec ies , and w i l l t o l e r a t e f a i r l y low
oxygen condi t ions (1-2 ppm).
Anbther spec ies , t he emerald' s h i n e r ( ~ o t r o ~ i s a t h e r i n o i d e s ) obtained
l o c a l l y from a Buffalo b a i t d e a l e r was used i n some experiments in o rde r t o
ob ta in a comparison of t h e t o x i c i t y t o f a theads and t h i s l o c a l l y abundant spec ie s .
The bio-assa;ys were jmde a t ord inary l abora to ry temperatures which were
wi th in t h e range from 22 t o 25* C . and would compare favorably wi th maximum
summer water temperatures i n t h i s a rea .
The t e s t s were designed so t h a t no oxygenation o r a e r a t i o n w a s gene ra l ly
needed, Absorption of atmospheric oxygen by t h e exposed water sur face was
normally adequate f o r f i s h requirements during t h e t e s t period. *However, i n
s o m of t h e bio-assays, high oxygen demand e f f l u e n t s caused oxygen dep le t ion .
When necessary, oxygen was maintained b y bubbling pure oxygen through t h e t e s t
s o l u t i o n by means of a s u i t a b l e arrangement of va lves and small tubing. The
r a t e (60-180 bubbles per minute) was ad jus t ed t o maintain adequate oxygen f o r
f i s h s u r v i v a l wi th minimum a g i t a t i o n t o prevent t h e l o s s of v o l a t i l e materials. I
Physica l and chemical determinat ions (temperature, d i s so lved oxygen, pHv
a l k a l i n i t y , and hardness) were made on each concent ra t ion both i n i t i a l l y , a f t e r
f i s h - mor ta l i ty , o r - a t t h e co@let+on oT t h e t e s t . . T h i s w a s done p r i m r i l y t o
maintain cont ro l of oxygen condit ions and t o d i f f e r e n t i a t e between f i s h
mortaITty due t o oxygen-defici.ency o r a c i d i t y and o t h e r t o x i c p roper t i e s .
F ish reac t ions were observed over a 96 hour period. From t h e mor ta l i ty
i n the d i f f e r e n t concentrat ions, 24, 48, and 96-hour TLm ( ~ e d i a n Tolerance Limit) -
values were obtained. The Median Tolerance Limit i s t h e concentrat ion of
e f f l u e n t i n d i l u t i o n water t h a t k i l l s j u s t 50 percent of the t e s t f i s h . These
values were obtained by s t r a i g h t ' l i n e g raph ica l i n t e r p o l a t i o n from percent 8 .
s u r v i v a l of f i s h and l o g concentrat ions bracketing t h e 50 percent point .
E f f luen t samples were n o r m l l y brought i n t o fihe l abora to ry i n the morning
immediately following c o l l e c t i o n and exploratory tests s e t up. Observations by
l a t e af ternoon indica ted the t o x i c range and f u l l s c a l e t e s t s were then s e t up.
A s some of the e f f l u e n t s were h ighly ac id and a t pH values,which would be
r a p i d l y f a t a l t o f i s h , similar bio-assays were s e t up with por t ions of sample
t h a t had been neut ra l ized with lime t o pH 7.
Bio-assays were a l s o made on mixtures of "A" ttB&Cn, and nEn e f f luen t s ,
mixed i n proportion t o t h e volumes re leased i n t o t h e Buffalo River,
Resul ts
Physical and Chemical C h a r a c t e r i s t i c s - A genera l desc r ip t ion and some of t h e chemical c h a r a c t e r i s t i c s of the
p lan t s rrAts, rtB&CM, and nEE'' e f f l u e n t s and the r i v e r in take waters a r e shown i n
Table 1.
A l l of these e f f l u e n t s were h ighly colored, with the i n t e n s i t y o r shade
varying somewhat i n t h e d i f f e r e n t composite samples. The "A" e f f l u e n t was
genera l ly from green t o blue, the nB&Cn e f f l u e n t from orange t o red, and the nEn
e f f l u e n t from pink t o d i r t y - o r a n g e . A mixturn of these e f f l u e n t s gave a dark
- 9 -
brown color, s imilar to, bu t of higher in tens i ty than, t h a t of Buffalo
River intake water.
Odors were var iable i n i n t ens i t y but some samples of "A" and "Elt
e f f luen ts smelled of very strong solvents (chlorobenzene, e t c .) . L i t t l e
o r no odor was noticeable i n the " B a f i eff luent . A l l e f f luen ts were
r e l a t i ve ly clear, containing only a small amount of suspended matter, o i l ,
or other extraneous material . Some samples, especial ly the tlEtl eff luent ,
formed prec ip i ta tes when added t o the d i lu t ion water.
Dissolved oxygen values were high in most IIAu and I'E" e f f luen t
samples and low values were ref lected only when r i v e r intake values were
a l s o low. Evidently organic materials i n these -effluents a r e very slowly
oxidizable. VlBgrCft sampl.es, however, were e i ther very low in o r f r ee of
dissolved oxygen.
With the exception of two samples of ltAtl, a l l e f f luen ts were highly
acid with pH values ranging from 2 t o and ac id i ty values from 100 t o 1200 ppm.
The "EN eff luent was normally the most acid, followed by "Ea" and then flAtt.
When neutralized with lime t o pH 7, medium t o heavy precipi ta tes formed in the
"B&Ctt and "E", but l i t t l e or none i n the "An ef f luents . A color change from
blue t o green was apparent i n "An samples when neutralized.
Phenols vere highest i n "B&C" ef f luen t samples ranging upward t o about
2 ppm with other eff luents containing generally l e s s than 0.5 ppm. Cyanides
were a t very low values i n a11 eff luents .
. TABm 1 - PHYSICAL AND CHEMICAL DATA
NATIONAL AIOILINB EITLUEETS
Dissolved Alkal ini ty Acidity Bardness hena ale Date Description ' Oxygen (C&03) (OaOO3) (CaC03) P P ~ Cyanide
Source 1956 - - Color,odor,etc, P P IPH PPm PPm PPm 4AA Gibbs - ppb -- Plant QA# 1-26 Dark purp l i sh-blue . 6.0 3.0 0 120 300 820 #3 < 5 Ef f l w n t Faint solvent.
. Slight turbidity.
24-Hr.Com- 2-15 Darkgreenish-blue. 8.0 2.8 0 325 175 24e 146 <5 poei te Burnt rubber.
-3
Slight turbidi ty .
I 2-16 Dark blue. 6.8 3 e 8 0 100 150 113 136
Burnt rubber. <5 P
0
190 turbidi ty . I
2-23 Dark bluish-green . 9.0 3 3 0 150 145 - g - Strong burnt rubber. BTo turbidi ty ,
3-14 Greenish-blue, 6.8 5.8 24 30 130 474 535 <5 Faint solvent, Slight turbidi ty ,
3-21 Bright green, 6,6 3.0 0 155 185 137 134 < 5 Sweetish solvent, No turbidi ty ,
'6-21 Dark green , 0.0 7.8 144 10 135 g - - Faint solvent. l o turbidi ty .
TABLE 1 (Contld) - PHYSICAL AND CHENICAL DATA
D i seolved Alkal ini ty . Acidity Bardneee Phenole Date Description WVZen (caco3 (CaC03) (CaO03) P P ~
Source 1956 color, odor, etc. ppm 2.L ppm ---- 4AA Qibbe
Same efflu- 2-15 Changedcolor fr,om 8.0 7.2 54 16 305 - - ente ae blue t o green . above. Eeu- No precipi ta te . t ral ized with lime 2-23 S l i a t precipi ta te . 8.4 7.4 58 20 330 - - Plant nBfBC" , 1-18 Dark reddieh-orange . 0.0 2.6 0 380 7 k 1026 1180 Effluent Faint solvent . 24-Hr .C om- Sl ight tu rb id i ty . poei t e
- 1-19 Dark red. - 2.4 - - - - - h i n t eolvent , Slight turbidi t y
2-14 Dark reddish-or ange . 2.0 2.9 0 660 200 1 9 3 1836 No odqy. No turbidi ty .
2-21 . Dark or ange-re d . 2.0 3.2 0 350 280 2091 2039 No odor. &o turbidi ty .
3-13 Dark orange-red . 0.0 2.9 0 810 480 1014 . 1071 Faint burnt rubber. No tu rb id i ty .
3-21 Dark orange-re d . 0.0 2.5 0 600 300 - - No odor. Sl ight turbidi ty .
Cyanide PPb
TABm 1 (Cont 1 d) - PHYSICAL AND CHFMICAL DATA
SAT1 OWL AN1 LINE EBgLUEBPS
p i ssolved Alkalinity Lcidi t y Jlardne ss Phenols Pa te Descripti on Oxygen (caco3 ) (CdO3) ( G a g ) P P ~ G yanide
source 1956 - - Color, odor,etc. P PH P P PPm PPm 4A.A Gibbs -- P P ~
6-20 Dark orange-red . 0.0 2.6 0 420 520 - 210 <5 Sweetish solvent. l o turbidi ty ,
Same eff- luent a s above. Neutralized with lime.
1-19 E'airly heavy broom - 7.0 - - prec ip i ta te .
2-14 Dark brown precip- - 7.0 - - i t a t e .
2-21 Small amount of brown - 7.0 - - precipi ta te .
3-13 Heavy dark brown - 6.7 44 18 prec ip i ta te .
P l a t ' IlEH 1-24 Light orange. - 1.9 0 770 240 864. 896 <5 Effluent Strong solvent 24-~r . Cow (chlorobenzene) . pos i te No turb id i ty .
2-15 Light pink. 9.4 3.0 0 28 0 150 774 739 (5 Faint solvent (sweet ) . No turb id i ty .
2-17 Light dirty-orange . 7.0 2.7 0 480 125 - 166 (5 Taint solvent (sweet ) . Slight turbidi ty .
2-21 Light t a n . 10.0 2.5 0 1020 135 461 422 <5 ra in t solvent (sweet ) . Slight tu rb id i ty .
T A B U 1 (Cont 1 d ) - PHYSICAL A I D CHEMICAL DATA
. Diesolved Alkal ini ty Acidity Hardnese Phenols D a t e Description 0 4 g m (Oaco3) (CaC03) (CaC03) P P ~ .Cyanide
Source 1956 Oolor, odor, e t c. - PPm pH PPm P P s PPm 4M Gibbs - -- P P ~
3-13 Dirty bluish-gray . 8.8 2.6 0 960 135 182 179 20 Strong so lventi (but y 1 aldehyde ) , Slight turbid1 ty .
Same eff lu- 1-24 e n t s a s above. NBUI t r a l i z e d 17 wfth lime
Pink, 4.4 Solvent (sweet ) . 190 tu rb id i ty .
Pink . 10.4 Strong solvent (sweet ) . No turb id i ty .
Light dirty-orange . , 0.0 S o l v ~ t 0
No t u r b i d i t y o
Light colored pre- - c i p i t a t e . Fa i r ly heavy l igh t 6.8 colored prec ip i ta te .
Heavy 1 i&t colored 7.6 f locculent precipi- t a t e .
3-13 Dark brown flocculent - 7.4 90 0 1130 - prec ip i t a t e-set t l e s to c lear solution.
TABXI 1 (Oont Id) - PHYSICAL AlVD CHIWICAL DATA
XATIOHAZI ANILIrJBI EmO%NTS
.Dissolved .Alkalinity Acidity .Hardnee8 .Phenol8 S a t e Description O X Y ~ ~ (Ca00,) ( a d o 3 ) (CacO3 ) P P ~
.Source 1956 - Color,odor,etc. P P 9H PPm PPm P P 4M Gibbs -- Mixed nAn, A -3-14 Dark purplieh- 4.0 2.6 0 320 230 - - MB& n, and BBbd -3-13 brown. nBIn efflu- 8 03-13 Sweetieh solvent. ents. I n Slight turbidity. proportion to volume A,B8C, Dark brown color. 4. 4 2.6 0 h 0 28 0 - . - releaeed. and 8- Solvent odor.
3 - a Slight precipitate.
A 6-21 Dark red-orange. - 5.5 . 24 100 325 - - B&-6-20 Sweet i sh eolvent . B 6-20 Slight turbidity.
River In- 1-18 Brown. 8.0 6.7 72 30 216 651 980 takewater . Faint o i l 6 sewage. 8-Hr. Com- Slight turbidity. -
poeit e 2-14 Light grayish-tan. 11.0 7 -3 80 6 135 92 79
Faint sewage. Slight turbidity.
2-16 Light grayish-tan. 7.6 7.6 64 4 110 52 29 Faint sewageo Slight turbidity.
2-21 Light grayieh-tan. 9.0 7.5 66 6 125 70 73 Oaeoline & eewage. Slight turbidity.
!PABGE 1 (Cont 1 d ) - PEKSICAL AND CHEMICAL DATA
-Dissolved .Alkalinity Acidity 3ardness Phenols Date Description OJWgen (0aC03) (CaC03) (CaC03) PPb Cyanide
Source 1956 - Color ,odor, etc. PW fl PPm PPm PPm 4M Oibbs -- PPb
3-13 Light grayish-tan. 10.0 7.5 72 4 115 11 10 5 Faint gasoline. Sl ight turbidity.
3-21 Clrayish-tan. 10.8 7.2 8 6 Paint gasoline. Sl ight turbidity.
6 Dark brown. 0.4 6.8 72 Faint solvent. S l i & t turbidity.
Toxic i ty t o F i s h
A summary of bio-assay r e s u l t s showing t h e d i r e c t t o x i c i t y of t h e
e f f l u e n t s t o f i s h i s shown i n Table 2 . The i n f o r m t i o n on which these r e s u l t s
a r e based, inc luding concent ra t ions t e s t e d , time f o r f i s h mor ta l i t y , percent
of mor ta l i ty , and bio-assay c o n t r o l d a t a are'shown i n the Appendix. . .
Median to l e rance l i m i t (T4,) values were obtained f o r 24, 48, and 96 - hour per idds . The 96 hour T4, was used t o compute t h e d i l u t i o n . r a t i o , which - i s the r a t i o of a u n i t volume of e f f l u e n t t o d i l u t i o n water which produces a
50 percent m o r t a l i t y of t h e t e s t f i s h . This r a t i o - t imes t h e e f f l u e n t f low
g ives t h e d i l u t i o n volume o r t h e t o t a l - f l o w o r volume o f rece iv ing water
requi red t o reduce t h e t o x i c i t y of t h e e f f l u e n t to where 6 50 percent
m o r t a l i t y of t h e t e s t f i s h i s produced. The fol lowing formula hay be used
f o r d i r e c t l y computing t h i s d i l u t i o n volume:
x E f f l u e n t Flow = Di lu t ion Volume
The d l l u t i o n volumes r ep resen t coriai t ions which would cause d 5 r e c t
i n j u r y t o f i s h upon s h o r t t ime exposure and a r e a d i r e c t comparative measure
of t o x i c i t y . L i b e r a l a p p l i c a t i o n f a c t o r s must be app l i ed t o t h e s e r e s u l t s f o r
complete p ro tec t ion of a q u a t i c 1Ffe.
A l l samples of nAlt, nWn, and e f f l u e n t s were t o x i c to f i s h . bAn
and "EtI were about equa l ly tox ic and ng&Ctl cons iderably l e s s . The t o x i c i t y was
v a r i a b l e wi th 96 hour TLm values ranging from 4.2 t o 15 percent f o r t h e "An -
e r f luen t , 13.5 t o 28 percent f o r the nB&Cn, and hi0 t o 24 percent f o r t h e I1Eff
e f f luen t . Intake water samples were r e l a t i v e l y non-toxic, a s i g n i f i c a n t
mortality' of f i s h occuring only i n t h e June (low flow) in take sample.
The -d i f ference -in 24 and -48 -hour TL values f o r the "An o f f l u e n t was s i g n i f i- m -
c a n t l y g r e a t e r than f o r t h e others , which may r e f l e c t some chronic o r accumu-
l a t i v e tox ic i ty .
Neutra l iza t ion d t h lime did not s i g n i f i c a n t l y reduce the ' t o x i c i t y of
the "An o r "En e f f l u e n t s bu t considerably reduced that of the " B a n e f f luen t .
In some of the samples, t h e add i t ion of d i l u t i o n wa te r t o the raw e f f l u e n t
r a i sed pH values t o *safel1 l e v e l s and yet a rapid mor ta l i ty of f i s h occurred.
Neutra l iza t ion would not be expected t o reduce t o x i c i t y very much i n these
s i t u a t i o n s .
Some d i f f i c u l t y was encountered i n maintaining adequate oGgen i n the
high concentrat ions of neut ra l ized "B&Cn e f f l u e n t . Whether f i s h l o s s was
due t o oxygen deple t ion o r t o x i c i t y could not be c l e a r l y diAtinguiqhed.
This d i f f i c u l t y was a l s o encountered i n a few samples of the "A1! e f f l u e n t . I n
most samples of f lAw and nE1l, oxygen was adequate f o r f i s h su rv iva l without
maintaining it a r t i f i c i a l l y .
The ef f luents were more toxic t o sh iner? ( N o t r o ~ i e a ther inoides) , a
l o c a l l y important forage species, than t o f a thead , minnows.
Mixing the "An, nB&Cn, and lbEV e f f luen t s d i d not s i g n i f i c a q t l y a l t e r
the t o x i c i t y i n two of t h r e e samples. I n t h e o t h e r s t h e t o x i c i t y was reduced
by one-half .
TABLE 2 - SUMMBRY OF BIOASSAY U T A
EATIOIOAL AIPILIm EFB2UXl!l?S
affluent TLm ( ~ e d i a n Tolerance Limit ) Date Blm (Per cent concent rat ion)
Source 1956 cfe 24 hr. 48 hr. 96 hr. - Plant I A U 1-26 47.4 10 4.2 4.2 Effluent 24-hr, 2-19 n 20 16 15 Compoei t e
Smt r a l i zed 42 24 21
lPetlt ralized '24 - - 3-14 n 10 7.5 7.5
3-21 n 16 7 6.2
Shiners 7 4.5 4.5
Average 17.4
Dilution Dilution ' Volume ' Ratio. cf e
(1) Intake toxici ty subtracted.
TABU 2 (Cont Id) - SUMMARY OF BIUSSAY DATA
BlllTIOlsAL ANILIIIB BFFLUENTS
$f fluent TLm ( ~ e d i a n Tolerance L i m i t ) .Dilution Date Flow (Per cent Concent rat ion) Dilution Volume
Source 1956 cfe 24 hr. 48 hr. 96 hr. Ratio cfe - .Plant WJOI' 1-18 10.5 28 20 28 1 8 2.6 27 Pf f luent 24 hr. 1-19 I 24 24 24 113.2 33 C ompo e i tie
Neutralized 75 - - 2-14 11 2 l 19 18 114.6 48
Neutralized 65 42 - 2-21 It 22 22 16 115.3 55
Neutralieed >56 47 47
Neutralized >56 39 35
3-21 n 13.5 13.5 13.5 ig6.k 67
Shiner e 8 6.2 6.2
6-20 11 24 24 24 ~ 3 . 2 (1)
Average 10.5
(1) Intake toxicity subtracted.
- 20 -
FABLE 2 (Cant Id) - SUMMARY OF BIOASSAY DATA
NATIONAL ANILIm ETI'LUEN!M
~ f f l u e n t T L ~ (Median Tolerance L i m i t ) 'Dilution Date Flow (Per cent Concentration) Dilution Volume
Source 1956 cfe 24 hr. 48 hr. 96 hr. Rat lo c f s - piant VP 1-24 2.8 6.2 4.2 4.0 1:24 67 Effluent 24 hr. Beutraliz ed 7.5 7.5 5.6 Compo el t e
2-15 11 24 24 24 1:3.2 9
3-13 n 7.5 7.5 7.5 1:12.3 35
Neutralized >18 >18 >18
Average 2.8
Mixed flA1', A -3-14 n w y , md m-3-13 30.7 Q t t ,
9 93-13
aff luent e 3-21-56 n 13.5
In propor- A -6-21 . t ion t o BM-6-20 11 13.5
E -6-20 volume released
(1) Intake toxicity mbt rac teb
'eWmTJ3e Lq pepnqyrquoo p q p iuymrepep op s p u e n ~ j j e prra~d r o j j s q p m u j pepos~pqne s 8 ~ m ~ o a uorpntra apuw~d Lq peen ~ e p 8 ~ r e a y ~ (6)
'p@W=Jlqne &rap09 @qeWI ('I)
9 * u h s $ 0 0 ~ * u n e @or *AJ- @or 9 r-z epyeod 4 0 9 *Jq-Q
- *Arne $ 0 0 ~ *Arne $00~: * m e @ot M qr-z J@ %ah 64e In1 - ( z ) * ~ . m $OL me $06 '-0 $ 0 0 ~ L*o€ 8‘1-r =ATU
s igni f icance of Data
Charac te r i s t i c s which Affect Aquatic Life - While bio-assays have shown p lan t ffAr1, "W", and "Eft e f f luen t s t o be
toxic t o f i s h , i n s u f f i c i e n t inTormztion %as ava i l ab le t o i n d i c a t e the m j o r
components responsible. The highes t tox ic i ty ; however, was genera l ly i n
e f f l u e n t s with a s t rong solvent odor. The chemically complex nature of
these wastes and t h e numerous processes from which t h e y ; a r e derived may make
it q u i t e i l i f f i c u l t t o determine sources or t o x i c mater ia ls . Bio-assays would
be of considera%le -help i n t r ac ing t h i s t o x i c i t y t o process e f f l u e n t s o
Bio-assays conducted an phenol and 'or tho-cresol under experimental
--condi-tions similar .- to t h a t of - t h e -e f f luen t s gave 96 hour TLm values of 40 - and 24 ppm, respect ively . This would ind ica te that phenol was n o t t o x i c t o
f i s h a t l e v e l s found i n the e f f luen t s . Other phpnolic compounds, e s p e c i a l l y
some of the chlorophenols, have been r e p o r t e d t o x i c t o f i s h i n concentrat ions
a s low a s O 0 1 ppm.
While the quan t i ty and nature of t h e solvents i n these wastes a r e not
known, many hydrocarbon solvents a r e t o x i c to f i s h . Bio-assays with some
solventg gave 96 hour TLm values f o r benzene, 56 ppm, - f o r toluene, 51 ppm, and - f o r xylene, 28 ppm. ,
Sub-lethal concentrat ions of phenols a d benzene de r iva t ives , such as
chlorobenzene, a r e known to impart a t a s t e t o f i s h f l e s h o
Cyanide concentrat ions i n t h e s e e f f l u e n t s were a t l e v e l s which would
not be expected to e f f e c t aquat ic l i f e .
The high acia'ity and corresponiing low pH values or these effl.uents
would have a detrimental e f f e c t on aquatic l i f e , unless adequate buffering
capacity was ava-ilab>e i n t h e rece'iving water or neutra l izat ion was affected
before release. Normally, pH values below 5 may be expected t o cause f i s h
mortali ty. Even above l e t h a l levels , decreasing the pH may have considerable
e f f e c t on increasing the t ox i c i t y of ce r ta in materials, especial ly metal s a l t s
and su l f ides . For example, l a rger amounts of metal ions such a s copper,
lead, zinc, iron, etc., a r e i n solution a t lower pH values. A decrease i n pH
of 1 u n i t may cause a 10 t o 100 fo ld increase i n t h e t ox i c i t y of some metals.
Likewise, decreasing-the pH releases hydrogen su l f ide from su l f ides and the
t o x i c i t y -may be .-greatly increased. Cun~snt ra t iuns of H2S a s low a s 1 ppm
a r e known t o have an e f f ec t on aquatic l i f e o A reduction i n pH m y have an
e f f ec t not only on matiri'als released i n these e f f luen ts but on those
released by other .induskries and present i n the receiving waters.
Color, apparenEly, has no a i r e c t e f fec t on f i sh . No re la t ionship
could be established between color in tens i ty and tox ic i ty i n any of the
eff luents . An ind i rec t e f f ec t would be possible by a dacrea.se i n l i g h t
penetration and consequently the productivity i n receiving waters.
Oxygen depleting charac te r i s t i cs may have a de f in i t e effect: on aquat ic
l i f e unless adequate volumes of receiving water a r e avai lable f o r d i lu t ion o r
assimilation. Dissolved oxygen concentrations should be maintained a t 5 ppm
or above i n receiving waters f o r complete protection of aquatic l i f e .
The re lease of l a rge volumes of e f f luen ts a t high temperatures which
increase those i n receiving waters may have an e f f e c t i n increqsing the
t ox i c i t y of some materials, reducing dissolv?d oxygen, and a d i r e c t off e c t
on some aquat ic organibmso
Effec t on Receiving Waters - AT1 of -these e f f luen ts were toxic t o f i s h . The d i l u t i on volumes
necessary t o reduce t h i s t ox i c i t y t o SO percent mortal i ty of t he t e s t f i s h
(Table -2) would obviously not proviae Tor complete protection of aquat ic
l i f e . L i h r a l appl icat ion f ac to r s must be used, which a r e based on the
following major considerations:
(1) The bio-assay procedure measures 50 percent mortal i ty during a
r e l a t i ve ly shor t time period i n non-renewed solutions. .This must be re la ted
t o no e f f e c t from continuous long tik exposure.
(2) Test data apply d i r e c t l y t o the species of f i s h used. While fathead
minnows a r e of intermediate tolerance and compare favorably q t h many warm
water "game" f i shes , some-locally important species of f i s h and f i s h food r
organisms may'be more sensi t ive . Bio-assays conducted with a l oca l l y important
forage f i s h , t he emerald shiner ( ~ o t r o p i s a tber inoides) , indicated t h i s species
was approximately twice a s sens i t ive a s fathead minnows.
(3) Some efTluents may vary in t ox i c i t y , A few samples may give an indicat ion
of t o x i c i t y -but maximum contiitions may 'be missed , In many s i tua t ions , maximum
condition3 o'f t ox i c i t y ore t;he".limi.ting Tactor a s far a s aquat ic l i f e i s
concerned.
(4) Some conditions which may generally reduce bu t . . in some cases, increase the
magnitude of the appl icat ion f a c t o r a re the l o s s of vo la t i l es , hydrolysis,
ox'idation, precipi ta t ion, and changes i n water qua l i t y cha rac t e r i s t i c s by the
entrance of other e f f luen ts . I
Based on t h e b e s t ava i l ab le information an app l i ca t ion f a c t o r of a t
l e a s t 3 would be necessary t o prevent f i s h k i l l s and a s much a s 1 0 f o r
complete protec t ion and propagation of f i s h . It i s evident t h a t low seasonal
f lows in t h e Buffalo River would not provide t h i s necessary d i l u t i o n ; however,
only a small por t ion of t h e Miagara River flow would be necessary.
This conctribution of t o x i c i t y t o the Buffalo River adds tot t h a t present
from o the r i n d u s t r i e s i n the b a s i n , While t h e t o x i c i t y from ind iv idua l
i n d u s t r i a l ' ~ T f l u e n t s is subs ' t an t i a l ly reduced' 'in t h e basin, enough remains
to present a t h r e a t ' t o t h e Niagara River under abnormal condjtions. Toxici ty
bu i lds up i n t h e Buyfalo River during low3'lows and then may be r a p i d l y
f lushed i n t o t h e Niagara River. Addit ional d i l u t i o n water i n the Buffalo River
during low flow periods would minimize t h i s p o s s i b i l i t y . A s long a s l a r g e 4
q u a n t i t i e s bf unknown t o x i c ma te r i a l s a r e enter ing t h e Buffalo River basin,
it cannot be s t a t e d w i t h . c e r t a i n t y t h a t t h r e a t t o a q u a t i c l i f e i n t h e Miagara
River w i l l be completely el iminated.
While oxygen dep le t ing c h a r a c t e r i s t i c s of these e f f l u e n t s wqre somewhat
masked by h igh t o x i c i t y , .in t h e bio-ssays, dep le t ion of oxygen was 'observed in
nB&Cll and some "An samples. A s t h e volume of - these e f f l u e n t s may exceed flows
in the Buffalo River during low flows, oxygen dep le t ion would no doubt occur
in t h e r i v e r . This would no t s i g n i f i c a n t l y a f f e c t the Niagara River bu t may
have an f l r e c t i n a - b o r d e r zone where the ByfTalo River en te r s Lake Er ie .
The p o s s i b i l i t y . a l s o : e x i s t s .that i n .'off - 'flavor i n f i s h f l e g h may be
produced by small q u a n t i t i e s of phenols, solvents , o r o the r chemicals. . .
I ! Conclusions
Bio-assays have shown t h a t a l l samples of P lan t s nAm, s W ~ , and
"Em a f f l u e n t s were - toxic t o - f i sh . The t o x i c components were no t iddn t i f i ed .
River in take samples were genera l ly non-toxic o r had only a s l i g h t degree of
t o x i c i t y .
The 'IAW e f f l u e n t was t h e most s i g n i f i c a n t from t h e standpoint of
t o x i c i t y cont+ibdted t o -the Buffalo River. The "An and "En e f f l u e n t s were
equal ly t o x i c - b u t ' t h e volume of *An was much la rge r . The 11Bdd3 ! e f f l u e n t
was considerably l e s s tox ic . ,
NeutrQl iza t ion wi.t%.lime s i g n i f i c a n t l y reduced t h e t o x i c i t y of t h e
"B&CW e f f l u e n t bu t had only a s l i g h t o r no e f f e c t on samples of "Am and "Em
e f f luen t s . Most of the nAn and nEn raw samples rem-ined tox ic a f t g r d i l u t i o n
with..Lake E r P e . w t e r t o concentrat ions wi th pH values which would be %to le ra ted
by t h e t e s t f i s h . ',
Some oxygen dep le t ion was apparent i n concentrat ions of "BMW e f f l u e n t
during t h e 96 hour t e s t period. When t h i s e f f l u e n t was neutra l ized and
oxygen maintained, very l i t t l e t o x i c i t y remained. Nost samples of l1Av and
"Em e f f l u e n t , however, w e r e t ox ic i n concentrat iqns t h a t r e t a ihed adequate
oxygen f o r f i s h support.
These e f f l u e n t s a r e major con t r ibu to r s of t o x i c i t y t o thq Buffalo River I I
and add subs tmnt ia l ly t o that contr ibuted by o ther i n d u s t r i e s i h thq basin.
Seasonal low'flows i n t h e Bu'ffalo River do not provide f o r adequq te 'd i lu t ion of
these wastes. Under normal-flow conditions, however, l i t t l e o r no e f f e c t on
aqua t i c l F f e i n t h e Niagara River would be expected.
The buildup of t ox l c l t y i n the BGTTalo River b a s i n during low
flows, with ths poss ib i l t ty of a rapid re lease i n t o the Niagara River
under abnormal conditions does.present soma th rea t t o aquatic l i f e i n
the Niagara . Additional d i l u t i on wiiter i n the ' ~ u f f a l o River during low
flows would minimize t h i s poss ib i l i ty I
A s long a s l a rge quant i t i es of unknown toxic materials a r e entering
the Buffalo River basin , t he th rea t t o aquatic l i f e in the ~ i a ~ o r a River
cannot be coingletely eliminated. Bio-assays could be used t o t race sources
of toxic materials within a plant which would3e of help i n fu r the r reducing
o r eliminating the major toxic wastes.
Pweible lw D.O.
Burnt rubber o b r
~h
I
( & d i m boleranee
i
~ i m i t ) - PQ cent concentration
I - 21 - 21 - 42 - 24
Dark blw-'peen h t rubber o&
Slight pteoipitnte when m t m l i r e b .
brer comentmt ion
Bo rednotion In t a d c i t y imiimtma f r o m neutralination.
~~~ TABLE A3 -BIOASSAY DATA T m Ilss - rABlBID UIIBOYS W I O U AEILIHE DIVISIOE. AUIED C H S f I C b l AND DT8 C O R F ' ~ T 1 ~ DIWTIOB MA= UXE m L BWALO, rn TOBg
S O U R C E rna C O N C E R - N U M B E R VOLIIME T I M E O F D E A T H P E R C E N T S U R V I V A L C H E M I C A L A N D P H Y S I C A L D A T A
C O L L E C T I O N O F COLLECTlON T E S T E D T E S T S A M P L E 2 9 9 8 7 2 96 T ~ ~ E T E M P . DISSOLVED T O T A L VERSERATE loaL ALK A C I D I T Y ~ R D R E S S
R E M A R K S
P E R C E N T F I S k F I S L 1 F I S H F I S H H O U R S H O U R S H O U R S OC OXYGEN ppm.
S A M P L E (CiCp:3) ( c a c o 3 ) p p n ppn.
,Pbmt A 100 Wf lusnt P 130 Ormieh-blue 98lnt molvent odor Blight trubibity
Black preelpltate h mte? add&
48 hr. D.O. - 3.4
24 hr. D.O. - 2.4
i?b hr. D.C. - 6.0
8081~ r d n e t l m in toxicity w h e n ovgen m l n t a i n d
S l w t turbidity
Sam precipiLatlon rben neutralised
D u k rsd-oram
eilgbt turbidity
46 hr.D.9. - 1.4 Asnrted after 48 hr..
Aerated after 48 hr..
A P P ~ ~ TABLE B2 -BIOSSAY DATA ~PB(FP PI= - OA!BUU UHHWS BaTIOlJaL AEILXE DIVISIOE, ALLIED WBIICAL A D DYE COBPQ3ATIo8 DILUTIQB k%T= - ULHB B r n a L O , Bay T m x
SOURCL A N D CONCEl i - NUHaER VOLI l t tF T I M E OF DEATH I PERCENT SURV I V A L C H E M I C A L AND P H Y S I C A L DATA METHO? OF 'ITE O F T R A l I O N OF OF T E S i
F I R S T HALF O F A L L OF 2'4 '4 8 72 96 TIME TEMP. DISSOLVED C O L L E C T I O N OF T E S T E D T E S T S I M P L E
F I S n F IS l l HOURS noURS OC OXYGEN ppn. T O T A L VERSEMATE R E ~ A ~ ~ ~
AC 1 D l TY MRDIIESS SAMPLE PERCENT F l S h L I T E R S (C$:!) ( C a C q ) p ~ n
Blflutmt 7.0 h k brom Recipimte frmosd whfm mtral lred
h a t e d aft- 24 LPm.
Ananted after 21) be.
mrk o m - r e d
9-11 aDWIlt or bran preoipitate
br 48 hr.D.0. - 0.6 Anrated after 48 hrs.
72 hr.P.0. - 2.4
Poeeible l o r D.O.
2b hr9.0. - 2.1
mrk ornwe-red Brsetleh odor
48 hr. D.O. - 2.0 to 1 s a d m after 48 DW ' 24 h*. D.O. - 4.4
m e n nsutralited. l i g h t brom preclpi- w t e forms d C h se t t l e0 repldlg. Posaible lw 0.0.
Sl ight solvent odor.
Possible l o r pB
Dirty blue g n y . Strow wlvent odor. Sllght turbidity.
h k bro& f l o e d e n t precipitate d i c h s e t t l e s n p i b l ~ . n - i e i w may have baen reducod on
Sveetish solvsnt odor.
rtss pink preclflur i n all brer con?-
With mortality a* thie pH valaw,cum t n l i s o t i o n d d not decrease be erpectd toxicity.
~ 1 1 centretlons f i sh dled met 1n up. con-
Complete survival i n tb next lover con- centration (1.b) rodd e v e a Tlrm nlb of
Light dlrtg orawe Solvent odor S l u t turbidig
With mortality 0% pe 6 to 6.5. - trelicatian wnl4 not be ~ p e o t s d )o
n t h c q k e - . ~ h taiv at p~ 6.2. aeutralirntim rotrld not be erpected to reduce toxloity.
Brk red-orange Swetish solvent odor
Possible 1 4 D.O. via morwity at pa 6.8.neutraliration auld not be expected to reduce toxicity.
NP&mU TABLE II-1 -BIOASSAY DATA TEST PXSE - ?ATHEAD ~ ~ ~ 0 1 9 rumom ~ L I I I B DIVISOB. ALLIEO QIWU ABD mn c o a ~ m ~ ~ o n D'm'On Lm - EB'*
BUBBAW, BEI rm C-
SOURCE I N @ C O I C E N - NUMBER VOLUME T I M E OF DEATH PERCENT S U R V I V A L ' C H E M I C A L A I D P H Y S I C A L DATA METHOD OF OF T R A T I O N OF OF TEST
F I R S T H A L F OF A L L OF 2 4 C O L L E C T I O N OF
cOLLEcTlON T E S T E D T E S T SAMPLE U 8 7 2 96 TIYE TEMP. DISSOLVED T O T A L 'ICRSENATE TOTAL A C I D I T Y YRDE~S
REYAR1S
SAMPLE PERCENT F l S h L I T E R S F I S H F I S H F I S H HOURS HOURS HOURS 'OURS OC OXY6EI ppl.
(C;C,::' (CaCq)wl pm.
Bioer Intake 1-16-56 100 10 20 40 me. - - 100 70 I n i t l a l 20 8.0 6.1 12 30 216 Bmrm. Pbint a i l and a w e
Slight t*lbiQ.
L*t g n y i a h (so. h i n t aeng8 *. rUi@at turbidity.
List gnyimb tan. mint aelbg. odor. S lUht t&i&Q.
Light grnyiah tan. Oanolin and acme
alight t ~ b i a i t y .
light Faint grayiah p e o l l n e t.n. odor.
al ight turbidity.
& P ~ I X TABLE -BIOASSAY DATA TES? FISH - ~ r h ~ d m u~miows AED SHIBEBS
UTIOHAL NIILIBB D I ~ S I O B . ALLIFS C~WICAL AND DYE COBPOR~TIOB DIUIT~OU WHI - IAXE a 1 3
BUPOU, m YOBg
S O U R C C A ~ D M E T H O ? O F
C O L L E C T I O H OF S A M P L E
River In take m i n t gasol ine odor S l igh t t u r b i d i v
Same am a b m e
Dark B r w n m i n t solvent odor Low D.0.
Same aa above. . Bo redraction i n tox ic i ty %%en oaygenatcd.
O A T E OF
3-21-56
C O N C E N - T R A T I O N
T E S T E D P E R C E N T
100
N U H a E R OF
T E S T F I S k
5
VOLIIIIE OF T E S T SAMPLE
1 0
T I M E OF D E A T H
F I S H
P E R C E N T S l l R V l V A L
H A L F O F
F I S H
-
2 s HOURS
100
REMARKS
Grayieh t an
A L L OF F l S l l
-
C H E M I C A L AND P H Y S I C A L D A T A
s 8 HOURS
100
T I H E
I n i t i a l
7 2 HOURS
100
TEMP. OC
23
96
100
DISSOLVED OXYGEN ppm.
10.g 7.2
ALK
(Ci:i?) g6
T O T A L A C I D I T Y
( C ~ G O J ) P P ~
1 2
VERSENATE HARDNESS
ppm.
155
Docket No. 54 Bio-Assay Investigations for LIC