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