WATER POLLUTION CONTROL RESEARCH SERIES © 12020 EXG 03/7)
OCL
ONMF.NTAL PROTI-XrnON AGKNCY
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WATER POLLUTION CONTROL RESEARCH SERIES
The Water Pollution Control Research Series describes the
results and progress in the control and abatement of pollution
in our Nation's waters. They provide a central source of
information on the research, development, and demonstration
activities in the water research program of the Environmental
Protection Agency, through in-house research and grants and
contracts with Federal, state, and local agencies, research
institutions, and industrial organisations.
Inquiries pertaining to Water Pollution Control Research Reports
should be directed to the Chief, Publications Branch (Water),
Research Information Division, R&M, Environmental Protection
Agency, Washington, D. C. 20460
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SELECTRD WATER
RESOURCES ABSTRACTS
INPUT TRANSACTION FORM
w
EFFECT OF CHLCtflHATIOii Oil SELECTED ORGANIC CHEMICALS
Barnhart, F,. L.
Campbell, G. 3.
Hydroaeience, Inc., "estwood, New Jersey
12020 EXG
/.later Pollution Control Research Series «ov~120S6~S{£, March, 1972, 103 p, 38 fig,
17 tab, 13 ref
Fourteen industrial oiganie chemicals were examined for their persistence through
biological1' treatment, either as the initial compounds, or as degradation products.
Semi-continuous activated sludge systems were employed. The ability 01' each of the
cher.dcals to participate in reactions with fr.3e chlorine was then determined in a
series of batch experiments.
It WQ.O jrOtjrdthe'C certain of the test compounds formed persistent degradation products
during treatment. Five of the initial compounds reacted readily with chlorine, under
conditions commonly employed in effluent chlonnation.
Five of the chlorination products were further studied in respirometer experiments to
evaluate their persistence in mixed microMal systems. Their toxicity to fish was
determined using the static bioassay procedure.
In the final-phase of the study-, a series of bench scale, continuous flow ecosystems
were established for the evaluation of longer tern effects of three of the chlorination
products. Several varieties of organisms, representing different levels in Vae food
chain, were studied. (Lowry,-Texas)
*C'nlorinatioi;, -Cnemioal Reactions, sLiodeGraci£tion, Bicassay, Activated kludge, Toxi-
city. Instrumentation, Ecosystems', Food Chains, Microorganisms, Laboratory Tests,
V.'vistewater Treatment
*Degradation Products, *Crg-:ciic Chenicals, *Respiro:aetcr Studies
05B, 0$i, 05C .
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Send To:
W A TLR F?f ^Ol. ^'"c". S SCI f. N T .f
L. S PL r-t f T ML \ : Of" rnCiNi
v\A^MI'-.»'.ION o c ro.'-io
INFORM A TtON CC'-J T Li:
_Tc.\as University, Austin
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THE EFFECT OF CHLORIMATION ON SELECTED ORGANIC CHEMICALS
by
The Manufacturing Chemists Association
1825 Connecticut Avenue, North West
Washington, D.C. 20009
for the
Office of Research and Monitoring
ENVIRONMENTAL PROTECTION AGENCY
Project S12020EXG
March 1972
For full-liy HIP ftiivrinti'iiilMit ot l)ocu:::r:il.«. I'.?. r.oviTiimt'iii 1'rInlliiK Ollliv. tVntMiielon, '.'.<'. ?>i|'i.' - I
Ifc
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EPA Review Notice
This report has been reviewed by the Environ-
mental Protection Agency, and approved for
publication. Approval does not signify that
the contents necessarily reflect the views
and policies of the Environmental Protection
Agency, nor does mention of trade names or
commercial products constitute endorsement
or recommendation for use.
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The resuJ. t.-; of this uuuciy i," c; Lc? tc.cl that chlorina cion of ef-
fluents contriving certain org-^nic civ. mica Is can result in
the forra'itiun o.1.' stable reacticn products, which may or may
not contain chlorine. IU was further sliov.'n that there com-
pounds exc.rcise a rctnrdar.t influence on aquatic life.
Fourteen industrial organic chemicals were examined for their
persistence through biological treatment as tnc initial com-
pounds, or as deyrdCi.Titi.on products. Semi-continuous ?ctivated.
sludge systems were employed. The ability of. each of the
chemical.s tc participate in reactions with free chlorine was
then determined in a series of batch experiments.
Certain of the test compounds formed persistent degradation
products during f- rent men t. i?ive of the initial compounds re-
acted readily with chlorine, under conditions commonly cm-
ployed in effluent chlcriniition.
Five of the chlorinotion products were further studied in res-
piroiuetcr experiments to evaluate their persistence upon ex-
posure to a heterogeneous microbial population. Their toxi-
city to fish was determined using the static bioassay proce-
dure.
i
Finally, a series of bench scale, continuous flow ecosystems
was established for the evaluation of longor term effects of
three of the ch]orination products. Several varieties of 01-
ganisms, representing different levels in the food chain, were
studied.
This report was ju'... .tit tod in fulf ill-ten t of project ?;12020E>T,
under the partial sponsorship of the Knler Quality Office,
Environmental Protection. Agenny.
1.11
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CONTENTS
Section ' Page
I CONCLUSIONS 1
II RECOMMENDATIONS ''. "' 3
in. 'INTRODUCTION \ 5
IV STUDY OBJECTIVES \ 7
V GENERAL DESCRIPTION 'OF PBASE I 9
VI EXPERIMENTAL METHODS - PHASE I 17
Analytical Procedures
Biological Degradation Studies 18
Chlorination Experiments ; 19
VII RESULTS OF PHASE I '\'\ 21
/analytical Methods ;
Biological Degradation Studies 22
Acclimated Systems \
Unacclimated Systems ' 31
Discussion of Results
Chlorination Experiments .
Discussion of Results - Cirlorination Studies 54
VIII RESPIKOMETER STUDIES -PHAS3\II 65
Experimental Procedure -.1
Study Results \
t
IX STATIC EIOASSAYS - PHASE II ' 73
Series Number 'One
Series Number Two
Scricr, Number Three
Summary
PHASE THREi;
Selection of Study Compounds1
Flov; Throagh bioassay ftud.i-.2S
74
80
81
XI RESULTS CV" filASS III 83
Fisn :
Vascular Plants - 86
Benthic Macroinvertcbratcs 87
Microscopic Flora and Fauna. 89
Summary ' 99
XII ACKNOWLEDGEMENTS 10]
XIII REFERENCES 103
Preceding page blank
V
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riGLT.ZS
Number Page
, SCHEMATIC REPRESENTATION OF BIOLOGICAL TREATMENT ,0
PROCESS
2 BIOLOGICAL DEGRADATION OF ISOPROPANOL 24
3 BIOLOGICAL DEGRADATION OF ACETONE AND Mr.TIIANOL 26
. ( 4 CHROMATOGRAMS ILLUSTRATING m-CRESOL BIOr'RGRADATION 21
5 BOD PROGRESSION OF BENZOIC ACID 20
6 BIOLOGICAL TREATMENT OF Y-BUTANOL 30
1 VOLATILIZATION OF ETHYLBENZENE DURING OHLORINA- ,,
TIOU EXPERIMENT
; S REACTION OF CHLORINE WITH PHENOL 35
9 MOLAR CHLORINE UPTAKE BY TEST COMPOUNDS 37
C11RCHATOGRAHS OF PHENOL AND CHLORINATED PHENOL -_
SOLUTIONS
11 REACTION OF CHLORINE WIT-! m-CRE£OL 40
? CHROMATOGRAMS >,' ' m-CRESCL AND CHLORINATED 41
n-CREuOL SOLUTIONS
13 REACTION OF CHLORINE WITH HYDROQUINONE 43
ULTRAVIOLET ADSORPTION CPECTRA-REAC.TION OF . r
CHLO1-.'IN!E WITH HYEROQ'JIN'C.'.'E J
15 REACTION 0]' CHLORINE WITH ANILINE 47
ULTluY.'IOLET ABSORPTION SPECTRA. OF CHLORINE
.1.6 CONTACTED ANILINE SOLUTIONS: EFFECT OF 49
CHLORIiJi: DOGAGE
ULTi';AVIOLET ABSORPTION SPECTRA OF CHLORINE
17 CONTACTED ANILINE SOLUTIONS: EFFECT Of 50
CONTACT '.'.THE
18 REACTION OF CHLORINE KITH DIMETHYLAMINE 53
,q EFFECT OP 2,4,G-TRICHLC7.GPHENOL ON A MIXED
MICROBIAL 10PULATION
., EFFECT OF 2 , 4 , G-TP.ICHLC P.OPFiENOL ON A MIXED
MICROBIAL POPULATION
VI.
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FIGURES
(continued)
Number Page
.,, EFFECT OF 4~CHLORO-3-«ETHYL?HENOL ON A MIXED ,g
M1CROBIAL POPULATION
9? 'EFFECT OF 4~CliLORO-3-i-iE'.L'HYLPHENOL ON A MIXED f9
''MICRODIAL POPULATION.
23 EFFECT OF CHLORANIL OK A MIXED MICROBIAL 7_
POPULATION
?4 'EFFECT OF 2,4,6-TRiciiLORANii,iNE ON A MIXJ:U 71
MICROBIAL POPULATION
25 RESULTS OF STATIC BIOASoAY SERIES NUMBER ONE 75 .
I
2G .RESULTS OF STATIC BIOASSAY SERIES NUMBER ONE 76
/'
27 RESULTS OF STATIC BIOASSAY SERIES NUMBER ONE 77
nR MEDIAN TOLERANCE LIMIT DETERMINATIONS FOR
2,4,b-TRICHLOROPHENOL AND 4-CHLORO-3-METHYLPHENOL
25 RESULTS OF BIOASSAY NUMBER. 3 - P-BENZOOUINONE 79
30 FATHEAD MINNOW POPULATIONS versus TIME 84
,, BACKGROUND f-ACROIKVERTEBRATE POPULATIONS versus 00
31 TIME 88
^_ MAVIE-UP OF AVERAGE FOUR WEEK MICROFAUNA on
' POPULATIONS
33 M/'vKE-UP OF AVERAGE TWO-WEEK MICROFAUNA POPULATIONS 91
34 TOT; L. MICROFALNA POPULATIONS 92
35 STALKED CILIATE POFUL/.TIOiiS 03
36 MOTILt! CILIATE POPULATION'S 94
.37 DIATOM POPULATIONS - 95
38 BLUE-GREEN ALGAE POPULATIONS 96
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TABLES
Numb_cr ' Page
1 ORGANIZATION OF THE PROJECT 8
? CHEMICAL AND PHYSICAL CHARACTERISTICS OF TEST 10
COMPOUNDS 11
, CHROMATOGRAPHIC ANALYSIS OF TEST COMPOUNDS -,.
RETENTION DATA
I
. ULTRAVIOLET ABSORPTION CHARACTERISTICS OF TEST .,.,
COMPOUNDS
5 SUMMARY OF BIOLOGICAL DEGRADATION STUDIES 23
> SUMMARY OF COD DATA FROM BIOLOGICAL DEGRADATION _,
b STUDIES
7 RESULTS OF FUL'LTMINARY CHLORXNATIOH EXPERIMENTS 32
8 CHLORINATION OF PHENOL 36
9 CHLORINATICN OF m-CRESOL 39
10 CHLORINATION OF HYDROQUINONE ' 42
11 CHLORINATION OF ANILINE 46
12 CHLORIN7VTION OF DIMETHYLAMINE b2
13 PROBABLE PRODUCTS OF CHLORIlvATTOK 62
14 RESULTS OF STATIC BIOASSAYS 80
15 TABULAR SUMMARY OF FATHEAD MINNOW MORTALITIES 83
CONCENTRATIONS (mg/1) OF SUBSTRATES DURING VARIOUS Rc.
TEST PERIODS . .
17 VASCULAR PLANTS GROWTH 86
Vlll
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SECTION I
CONCLUSIONS
Phase I: Chlorinatior. Studies and Analytical Methods
!. Gas chromatographic analysis was successfully employed for
the detection of most of the initial test compounds in
aqueous solutions. The method was applied to the analysis
of effluents from bench scale biological systems, and was
user'l in monitoring the test chemicals during chlorinatior.
experiments.
2. Ultraviolet absorption (UV) spcctrophotonetry demonstratec
applicability to the measurement of aromatic compounds in
relatively pure aqueous solutions. The ir.ct.hod was usi.d in
compound monitoring during the bioassay experiments, and
provided information on the nature of the products result-
ing from chlcrination.
3. , There is evidence to indicate the formation of degradation
products of severs,! of the test compounds, during biologi-
cal treatment, j.n acclimated systems.
4. Five of the fourteen chemicals selected for study were ob-
served to participate in reactions with free chlorine.
under conditions encountered during conventional effluent
chlorination practice. The ability of these compounds to
react with chlorine can be related to the structural char-
acteristics of the chemicals.
5. Evidence indicates that chlorine reacts with these organic
chemicals by both substitution and oxidation, resulting in
a highly co.nplex mixture of products. In some cases, i'_
was possible to identify the products of reaction.
6. The nature and distribution of the products of reaction
with chlorine are affected by a variety of parameters,
including concentrations and contact time.
Phase II: Chlorination Product Persistence ard Toxicity
7. Several of the reaction products identified in Phase I
were examined in respiromoter studies ani found to be
resistant to degradation upon exposure t? a heterogeneous
microbio.l population.
8. Toxicity to fish by these -products was demonstrated in
static bioassay experiments, and 96-hour TLrri values were
established.
9. On the basis of tho results developed in Phases I and II,
it is evident that chlorination of effluents containing
certain organic chemicals nay result in the formation of
persistent and not.entiallv deleterious reaction products.
i
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rh.i.-.e III: Int jrmed: a_te Bicassay
1C. Considering the ^-dative efforts? involved, t.'ie intermed-
iate terra, flow through bioassays a:-; conducted did not
provide significantly Letter insight into the toxic ef-
fects of the test compounds, than did the rout: no four-
day static bioassays, The static to-st appears to pro-
vide a conservative tcxicity lir.it "ic.n minimum eforl.
11. Although some qualitative judgments of compound effects
on the test microorganism populations can be drav:n, the
data were toe variable to allow quantitative statistic.?.]
' determinations of toxicity. Qualitative evidence was
' suggestive of toxic or inhibitory effects on stalked
. dilates and diatoms for each of the three compounds at
. : the highest concentration levels tested.
.12. 7i significant increase in effort, well above that practi-
cal foe a project of this nature, would be needed to pro-
vide statistically definable: determinations of toxicity
to "r.ioroorganisius .
13. Of the two types of vascular plants tested, one exhibited
, an erratic growth pattern and was unsuitc'.ble as an indi-
/' cator of toxicity. The second type was not erratic in
growth, but no toxicity was observed. The general out-
ward appearance of all plants was not adversely afrectud
by "any of the compounds.
14. Stable macroinvertebrate populations could not be main-
tained in the model ecosystems for a time period of
suitable duration for compound testing.
15. On the level of experimentation performed, the energy and
food web interrelationships within the cxocr irr.cn tal cco-
svstems could not be defined.
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SECTION II
RECOMMENDATIONS
1. The practice of routine chlorination of industrial efflu-
ents should be re-examined. In cases where it has been
demonstrated or suspected that organic chemicals are re-
acting with chlorine to produce undesirable products, an
alternative disinfection mothod should be considered.
2. Effluents known to contain chlorine reactive materials
' j should be monitored for those materials, and for unusual
increases in chlorine de/nand.
3. The present study should he extended to provide further
information regarding the nature and properties of the
products of reaction between chlorine and industrial
chemicals, which have escaped biological treatment.
4. Further studies should be undertaken to determine the
effects of theo«? products of chlorination on biological
i systems. These studies should include evaluations of
fish toxicities.
5.: The intermediate term, flow through bioassay such as cm-
ployed herein, is not recommended for future investiga-
tion of compound toxicity. By comp.v.-.son, the routine
four-day scatic bioassay provides a simple method which
yields a conservative determination of toxicity to a test
organism, at considerable saving of effort.
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STCTIOM III
i:.'TRODuT7IOM
The chlorination of raw or treated domestic was tewate rs is
conifionlv practiced to achieve disinfect inn and deodorination.
In many municipalities, effluent chlorination is mandated bv
public health codes. In recent vears, this eountrv has wit-
nessed a trend towards combined nunirinp.l-industr.-i.nl waste
treatment. Additionally, an increas.inn number o ^ industries
have been required to provide waste-water treatment- faoilitios,
which often include.s eff'ucnt chlorination. 'p'-ios" dc^elo^m
have raised carious questions reoarrii. an t-.hr- irv.iot o f certain
industrial chemicals on was tewate1' tr^atren'-. nlant operation
iii general, and on chi.orir.nti.cn in i.art iruT nr. It i.s ooncei"-
able that, under eerta: n conditions, some oraanie comoou-ids
may escape treatment or be onlv partial Iv tlenraded, si.ich that:
they are a\'aiiable for reaction with chlorine in th^ contact
c!iantbcr.' Furthermore, tlic products of such a reaction could,
upon uischnrru.-, exercise a deleterious effect on the reeei^inn
sit rears.
A typical example of such an undesirable reaction is the com-
bination of chlorine with phono?, to produce the nh lornr-ilienols .
These materials are a source o1' obnoxious tastes am.1 odors,
even at very low concentrations (;.n/l) .
More-over. :>tud.ies bv Inools and Jacobs ' have s'rv^wn that the
ch] oroplionols are noro :."c.~ ic. tnnt to b ioder:rat:'.on t'ian !>honoi
itsoif. Trichlorophonol v;as cite:'- as boino toxic to ohenol
("')
ada-Jted microorqanisms . Chambers, ct.al. " also reported on
the increased resistance of the chlorinated uroriucts of both
phenol and ir.-crc.sol to bd occnradation. In an extension of an
earlier \/ork , In no Is; , et.nl. IJ d.eter-ir.ed that the c!iloro'.vne-
nols were more toxic to fish than phenol.
The precec'-'linn cf./.s ir.erat ioj'iS c!earl\r ip.r'i rate the need for a
re-exami:. it ion o ' e ' K\ v.er.v. ch l^'-ina t ion n r a c t i <:?. In addition,
rcsea)'c:i (..-^fort's ^h'lv..1 C. bt.: ciuected to'.-pj-dn id^nt iCi/i. nc: thoso
chemicals v/hic'-i -'.re ;.ir.~se".u ir ^'ac '-':>'o°r oirr'!.Monts and '-hirh
v/ill reart wiMi chlo7"d:.e. Ta"oi-:"Tt' on s'-io1.1.'1'1 >.TPO be d^vr-1 oned
jn. tiie 3 ;..:>act D': tiiope ma~cr i.als i^n ..lie er-^i OT-.- o1" a rereiTTin<^
'.; a t or.
In rerocn i t.ion o" these p?vh'lors , the ^'anu ^act ur i no rho^istR1
Association, ir. coo^-^rr" lep wi/h ':ho V-;:iv\rop~r.;-.<-_T'I ^roterti.on
A nor. t.".', in: t .La ::ed thi o:--.'r-ep.'j -tirh" ''or the r-::r::o-^. c c rrnth^j
ii'ig ir: f orrvi t.i.o:i 0:1 t'u.' <.; I'fe.Tt o '" ch j or1" :'.a ti^n on. certain in-
dus tri al ,ji\~:an i c comnov.r.ds .
Preceding pap blank
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SKCTJO:; i\-
s TP ny on J1": CT F " ' .c
The presort study war-; nndortn!:an in an attest to dovpl.no in-
formation on the ejects of chlori nation on selected ornanio
chemicals. The nood.to -levclon tostino prnoodrres and analv-
ticnl me'iihod.s for the characteri/atinn of tho--:c efforts './as
ju'dqod tr> he of pnranount -inportanc-c;. Specific nrc-]cct ob-
jectives included:
1. The nxani nation of the: i:i ^luonce of
selected oroanic copoounr's an '"'/or thoir
deoradat j.on producer; on chlo.vine de
2. The- identification o* those chenicals
whicii forr1 stable react.ion croquets uoon
contact with cli.lorine, under conventional
conditions of c!ilorinatinn.
3. The cliar.icter.iza tion of tho^e reaction
products in terrir, of their persistence
in biolonical svscr-ns, or their uotentia.l
to act o^ inhibitors or toxicants to svich
4. The deternination oc the effects o f
any persistent rei.c-tion nrodi!^tr- on the
ccolooy of a sir'ulated recei^'.ino stream.
Fron an orqani?.ationa]. standpoint, the studv '-/as arranoed into
three distinct phases, as 'nhov.'n in Tab]e 1. Kach successive
phase was. intended, to rcoresenc a Ionic a 1. invpstinati^e nro-
gression in the accor'olishront of tlie ohiocti^es of the nrolect,
It was envisioned .that sone effort in each of the nhascs would
proceed concur.ront.l-v , since th.c nature o^ the studies necessi-
tated the refincr'cnt or development o.f exne^i'^ental an.'i ana.l\'-
t.ica.l pothridolorrics . Consenuentl\' , su'-i^eenif^nt descriptions of
the studies are not intended to }~>c eh renol '-.: icallv consistent.
Preceding page blank
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1AELE 1
ORGANIZATION OF THE PROJECT
Chemicals ."elected For
__ atucly
Alcohols
Me t ha: 10 1 (1°)
Isopiropanol (2°)
T-Butanol (3°)
Acetone
Benr-eno a_p.d_ Derivatives
B(_T.^c-nr
Toluene
I'.Lhy [.benzene
Bcnzoic Acid
Phenol ur d I'honolj rs
Phase I
Phase II
Phase III
Phenol
m-Crcsol
llydroqiiinonc
Organic Mi t rcxjon Compound:
Aniline
Dime thy lam J no
Mi trobrnzcno
All compounds tested
l:or :
1. j.-uiLability of
analytical methods
2. persistence of
parent or degrada-
tion product
through biological
treatment
3. reaction with
chlorine
4. identification and
characterization
of products
Five compounds selected
on the basis of Phase I
experimental results:
1. evaluation of resis-
tance to biodegrada-
tion
2. determination of
toxicity
Three compounds selected
on the basis of informa-
tion ciathercd in Phases
I and II for evalua-
tion of effects o;; a
simulated ecosystem
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SECT 10?!
Phase I v/as -''_ voted to tho c:? the sta.rt.inr:
conpourids to carbon dioxide, water, new mi c rob in. I cells, and
possibly, sor".e interncdiate cop pounds ; or (?.) incornlete de-
gradation or no degradation, such that the starti.no rornnounds
appear in the effluent. In continuous 'svsters , SOPO residual
organic material will always remain in solution as a result or
an equilibrium bof-./cen the microbial cells and their liquor.
The compositions of treated effluents are hiohlv comnlex, and
have been chnracterized onlv in t'.:rrs of oeneral classes of
materials, e.g., carljohvdrat.es, proteins, tannins, "fulvlc"
and "hiu-'ic:" acids The nature and di.s tribntj.on ..T(: snecific
components is kno.vn to !^c affected )iv selection o'" treatment
parameters, as well as by the composition oc the food stock.
A we.ll function: rrr continuous bioleniral svstom, re reivinn a
feed stock of uniform composition, v.-ill achieve essential Iv
complete conversion of Lh.e organic- mater ia] initi-il.lv i.irese.nt.
Such a system is said, to be a_cc lira tod , in contrast to a svs-
te:v, v:h.ich is unacnl imatcd . A biolocical oroc-^r.s v/hir'n is sub-
jected to internfiTtent or shoe!: loads mav be iri -\ccli.-vite-l to
certain cornonents associated v;ith these io.ids .
:\t t'.: is junct-.ure, it :s useful ti re vi " the conditions under
v;hich a ij.i.o ].>(.: Leal fr.'SiicM'1 ac'iieve" accl ir-'iti. ration . Th'is ria>;
bo vi sual i:-;of' b a con.sj Jems: i on o-" tii^ fieneral .1 7,ed b.i.olonical
u rcatrn-n t process, as illustrated in ''inure 1. It is noted,
that \'.\\c v:>:owf;h. of .1 picrobial. noirnlation (s.ludno) iiroi-irosse'5
<.;) .rouch se\'oral p!ir.-,i\°. unon rxnosi.'--!--- to a substrate: first, a
- 9 -
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Compound
Mcthanol
Isopropanol
T-Butanol
Benzene
Ethyl benzene
Toluene
Benzole Acid
CHEMICAL ANI
. Molecular
Formula t, . . .
Wciaht
CH,OH 32
C!'V-CH -C.T .
CH3
CH,-C-CH., 74
J OH '
Ml 73
i |C2H5 106
:- li -3 92
,.-~ ,COOH 122
:^ '!
..04
;- 09
.12
.11
.16
.13'
.12
TABLE' 2
3 PHYSICAL CHARACTERI,
D^1:L"g Solubility
P?^nt (mg/lOOnl)
64.65
82.3
108.4 9,500
80.1 82
136.1 14
110.6 ' 47
249 270
STICS OF TEST COMPOUNDS
BOD ,- A . A , *
Density Oxygen 5 . ___
(q/ml) Demand (ma/ma) , / *
0.796
. 0.785
0.801
0.879
0.867
0.867
1.266
(mg/mg)
1.50
2.39
2.59
3.07
3.16
.3.13
1.9f>
1 ~" " VITlCJ/ lUCj /
1.0 1.395
1.45 1.79
0 1.65
0 0.13
0 0.05
0.86 0,13
1.42 1.83
(mg/mg )
1.46
2.34
2.46
0.82
0.89
1.58
1.87 .
*T<'ctinicon AutoAnalvzer m'ocedure.
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TABLE 2
(continued)
CHEMICAL AND PHYSICAL CHARACTERIST
, , Molecular B°llin9 Solubility
Compound Formula ,.,;,., Point ,_/, ~is
Phenol
M-Cresol CH3
Hydroquinonc _
Aniline
Nitrobenzene
>
^__/ °H 94.11 182 6,700
!'' :i°H 108.13 202.8 2,350
X -., OH
^ ' 110.11 286.2 5,900
--' , NH
1| 93.12 IS'!. 4 3,400
^ 'j1; N°2 123.11 210.9 190
i ^^^^ ';
Dimethylamine CHINCH, 45.08 7.4 V.S.
Acetone CH,-C-CH- 58.08 56.5
O
ICS OF
Density
(g/rhl)
1.072
1.034
1.358
1.022
1.198
0.68
0.792
TEST COMPOUNDS
rTotal BOD, A. A.* Reflux
oxygen s COD CCD
Demand (mn/mg) / / \ / / \
(ma'nc-) (mg/mg) (mg/ng)
2.38 1.75 1.785 2.32
2.52 1.70 l..:3 2.31
1.89 0.75 1.83 1.66
3.18 1.8 1.67 2.47
3.C5 0 0.19 1.39
3.75 00 0
2.20 1.4 .1.13 1.80
*Technicon AutoAnalyzer procedure.
-------
UJ
h-
CJ
O
o
CO
CO
llj
_J
LD
/I.MTi-V. COO REMOVAL
CLIOCE C.'JOWTH
I i AG | I.CG c:;0'.viM i CLCLI::I:-:G o.;;0.v!ii 1
A B . C D
TIME
FIGURE
SCHEMATIC RLlv^SENTATlCN OF BIOLOGICAL TREATMENT PROCESS
-------
lag phase, characterized by adaptation of the culture from
the previous environment to the present; second, a period of
maximum growth under conditions v/here unlimited food is
available; third, a period of declining growth where food
availability becomes a .limiting condition; and finally, an
endogenous phase v/here., under severely limited food condi-
tions, cell" die and are, in turn, consumed such that the
mass population is reduced. In heterogeneous systems, this
process occurs within the biological culture for each sub-
trate as a subset of the overall reaction. In a system
which is unaccliiT.ated to a particular compound, the lag
phase associated with that substrate may be prolonged, and
may result in the passage of that compound through the treat-
ment process, without any significant degradation.
In the evaluation of the; effects of chlorine on organic chem-
icals subjected to biological treatment, two cases should be
considered: (1) an investigation of the possible reactions
of chlorine with the parent compounds which would persist
through unacclimated systems; and (2) the determination of
the probability of chlorine reactions with degradation pro-
ducts of the starting compounds, which v/ould appear in the
effluents from, acclimated-biological systems.
The case of chlorine reaction with possible degradation pro-
ducts was examined first. Requisite to the evaluation of
potential effects of chlcrination is the establishment of
the existence of significant degradation products which uni-
quely result from .biological treatment of any of the test
compounds. In certain cases, the presence or .absence of de-
gradation products can be inferred from a consideration of the
nature of the reactants and their known degradation character-
istics. A simple substrate, such as methanol, should undergo
virtually direct conversion to carbon dioxide, water,, and
cellular material. For more complex organic molecules, degra-
dation may be accomplished by a series oi' reactions, involving
intermediate forms. .
A series of experimental tests can be applied for the deter-
mination of intermediate product production. Ideally, the
detection of the intermediate product itself would be the
most direct approach. However, no single analytical techni-
que is universally applicable, and the selection of a method
must necessarily be based on some information about: the ma-
terial being detected. In the absence of direct experimental
evidence, indirect procedures are often employed in the deter-
mination of the presence of degradation products. Monitoring
of the effluent for tie disappearance of the parent compound
(by chrome.tographic or fjpectrophotometric procedure j, for ex-
ample) and for a decreuso in oxygen demand may provide some.
useful information. If the rate of compound disappearance
closely parallels the rcite of COD removal associated with
that compound, it may be generally asserted that no signifi-
cant: degradation product buildup has occurred. On the other
- 13 -
-------
hand, if a not residual COP is observed, relative to a control
system, intermediate product forr-ation mav bo takino nlaco.
The lack of adherence to .?;toichiom,..trv for cxv<-r-n utilizatir,n
may provide additional evidence for "rhe production of i'l.ter-
mediates. If the complete disanncarr ncr.- of a parent conuci'r.d
is accompanied by substai.'-.iallv less thar- thi ^retieal oxvqen
consumption required to produce CO , war.'hotocheinJ cal reaction, etc.) of
conpound(s).
The nature of the b.ioloqical treatment T?vocesn itsel^ is not
conducive to the naintcnar.ee of true stead"-statc cojiditions.
Transient changes in microbial pooulat.ion d-'narirs i^av vielcl
erratic results. Bachoround levels of residua], byproduct ma-
terial from cellular l"3.is (on do at: no us rosvir^t i.on) nnv obscure
tlie presence of specific intermediate coFivx-unds.
Perhaps even more pertinent to the evaluation of the orobabil-
ity of chlorine reactions with deqradati^n product?, is the
fact that the nature and, distribution of doo.rad.Tt-.ion oroducts
nay be controlled by the selection of trontnent pararieters.
As an exariple, ammonia ("!U) nav be reo-irded as a byproduct of
organic nitroaen degradation. ilov;evcr, b-.' aporonriate acVjust-
r^cnt of loading conditions (food-to-r-,v:;roor.-;an is;-' ratio) , the
conversion of ammonia to oxidi?.od riitrcqon forrs (N'O^ !JO_)
may 'be encouraged. r'rirthcrmoro, it is know'i that anmonia will
participate in roactior.s with chlorine, lerd.inrr to the forma-
tion of chloranines, but nitrate underaoes no reaction with
chlorine.
In summary, it may be concluded that:
- 14 -
-------
1. present methods for establishing the
existence of intermediate products
resulting from trie biochemical de-
gradation of specific organic chemi-
cals lack general applicability and
are of questionable validity;
2. the nature and distribution of degra-
dation products may be drastically
< altered by adjustment of treatment
parameters;.
i 3. due to recognized experimental limi-
tations, the more practical case for
investigation is the ability of chlo-
rine to react with specific organic
chemicals, which hav; escaped biolo-
gical treatment.
In accordance with the considerations detailed above, the ma-
jor effort of Phase I was devoted to the examination of the
ability of chlorine to react with each of the initial test
compounds. Test conditions were selected to simulate those
that would be anticipated for full scale treatment plants:
dilute solutions of organic compounds, pH ranges near neutral-
ity, conventional applied chlorine dosages and contact times.
It should also be pointed out that all applied chlorine was
in the form of free chlorine, since ammoniafree solutions
wore used. The principal criterion applied in the determina-
tion of chlorineorganic chemical reactions was thi.t of the
chlorine demand, as derived from residual chlorine analysis.
Chromatographic techniques provided the means for monitoring
the concentrations of test compounds during the chlorination
experiments and, in some instances, facilitated the identifi-
cation of reaction products. Supplemental information was also
gained by the use of ultraviolet absorption spectrophotometry.
- 15 -
-------
SECTIO!-: .VI
EXPERIMENTAL METiiODS - PHASE I
Analytical Procedures
In view of the need for monitoring of the test compounds
throughout the experimental program, a substantial portion of
the effort in Phase I was devoted to tht; dovelopr.cnt of ana-
lytical, techniques. It was recognized that any r.etihod worthy
of consideration should have general applicability to the
range of selected organic chemicals end their derivatives, oe
capable of selective identification o.:~ snecific components in
complex systems, and be suitable for quantitative analysis at
low (mg/1) concentrations.
Liqvid-gas chromatography was initially chosen for these in-
vestigations. The instrument used.was a Perkin-Elrner Model
881 chromatograph with flame ionization detector and linear
temperature programing. Heliun was employed as the carrier
gas, while hydrogen arid air were supplied to the flame ioniza-
tion detector. Chromatograins were recorded on a Leeds and
Northrup Speedomax W recorder equipped with a Disc integrator.
Five columns were employed for chromatoqi-v.pr.ic separations
during these studies, as listed below:
1. 6' x 1/8" (uncoated) Foropak Q,
100/120 mesh, packed i:i st;i:-.iess
steel
2. 6' x 1/8" (uncoated) Poropak S,
150/200 mesh, packed in stainless
. steel .
3. . 6' x 1/3" 15% K20M Carnov:ax TP/.
on Chromosorb V7 HDMS, 8U/100 mesh,
packed in stainless steel
4. 12' x 1/8" 15% K20M Carbovax TPA
on Chromosorb '.-1 HDMS, 60/80 nosh,
packed in glass
5. 6' :< 1/8" Chromoscrb 103, SO/100
mesh, packed in glass
In all cases, dual column systems were utilized to facilitate
baseline stability by compensating for colur.in "bloc-cling" ef-
fects.
For purposes of parameter optimization and calibration, stan-
dard solutions of each of the test: compounds v.'ere prepared in
distilled water. Where available, chro~ato.;:raphic or reagent
Preceding page blank
- 17 -
-------
grade chor'.icals wore used. Injection of sr.mr>los '-/as accom-
plished with n llanilton microlitor syrinao of suitable size
(1, 5, or 1.0 ',:1) . Calibration curves were cons truotod hv
plotting the product of the attenuation factor and nrei (rel-
ative Disc integrator in its) an a function of samole weioht.
Standard}, xdtion was chocked frequently durina the experimental
program to insure quantitative rcproducibilitv.
It becanc evident that chromatoaraphic teohni nu.->s, employing
direct aqueous injections of sanplos, were not applicable to
the separation and quantitative detection of sore o *: the oom-
pouncls under investinat: on. It therefore was decided to ex-
plore the use of HV spoctrophotometrv as an analytical tool.
The instrument used for those, .studies v;as a Porkin-Hlnor Model
202 double-beam ratio-rerordino spectrophotemotor. Standard
solutions of each of the compound?; of interest were prepared
in distilled water. Absorbance-wavolonnth recordinns '-/ere
obtained in the 190-35C nm region of the spectrum, us inn dis-
tilled water as the reference. Calibration curves wore ore-
pared by plotting absorbance as a function of concentration
at each peal: wavelength. Ultraviolet spectral scans f^r san-
ples dissolved in <~ solvent other than distilled water en-
ployed the appropriate solvent in the reference coll.
B i o I o g i c a 1 DC g r a da { ion Studies
A series of semi-continuous activat.~, the feed consisted of 100 nr;/l of
each test compound, expressed as Theoretical OXV--IT. Demand
(TOO) , in combination with 100 mo7l (as Con) or a svn~t.hpt.ic
"sev/age1', containing a mixture OF rn-'idilv dofsradablo organic
compounds and inorganic nutrients. A control unit, which re-
ceived 200 mg/1 (as COij) of tin.- synthetic mixture v/as also
maintained durina those studios. This synthetic mo<-"Ma --jas
prepared at froquont intervcils in accordance '-;ith t.iie ^ollo1-/-
inn formula:
-------
SYNTHETIC DOMESTIC WASTE COMPOSITIOIJ FOR A COD OP 100,000 MG/L
' Ingredient- grams/li ti-r
skim milk 48
peptone 43
gelatin 1C
soluble starch 32
urea 8
disodium hydrogen phosphate 8
KC1 " * 1'. 12
' CaCl., 1.12
MgSG^j 0.30
i Fe2(C04)3 . O.L'O
MK,C1 8
H
Following a two-week period of acclimatization, each ot the.
systems was monitored for reduction in soluble COD and for
disappearance of test substrate, where chronatonrapr.ic techni-
ques wer,e available. Analyses were performed immediate."'.;/ after
addition of t-«?st substrate and synthetic mixture, and at se-
lected /Intervals tliereafter.
I
In an aerated biological reactor, substrate remcval may also
occur by diffused air stripping of volatile components. To
investigate the significance of this phenomenon for each of
the test chemicals, a separate series of tests was conducted
using the same apparatus as previously described. In these
experiments, the biological culv.ure was omitted, and :>n aque-
ous solution of each test ch-_mi:;al was aerated, us inn an air
flow rate similar to that employrd in the degradation studies.
The concentration of each test compounds was monitored with
time, using chronatographic or spcctrophotonetric procedures.
Chlori.nation Experiments
To determine the effect of chlorine on each of tho selected
compounds, two series of batch chlcrinatior. experiments verr.
conducted. The first set oi tests were designed to identify,
in a qualitative sense, which of the selected chemicals wer>_
capable of reacting with chlorine, under conventional treat-
ment conditions.
Aqueous solutions, containing approximately 10 me:/! of each
compound, were prepared using distilled writer and adjusted to
p!l 7.4, with a phosphate (K2I!1 0..-IC11. ro ) buffer. Where rosr,-
ib].e, the concentration of test chemical was chachcd using
chromatographic procedures. A stock chlorine solution, hr.v-
ing an approximate con':entration of 1,000 no C!.-,/! , v.'as pj-o-
pared from a commercial grade oi sodium hypochloritc. T'.is
solution was standardized frequently by iodir ^tric i:itra"ion
in accordance with the orocecures described in Standard :'ot!-.ods
- 19 -
-------
Each tost run consisted of four (!) 500 r:l sa~r>~'.es containinn
the tost compound, to which vnrvinn dosanos op chlorine were
applied. Since a gradual loss of chlorine? witn tir>e had boon
demonstrated in prelir-inarv tests, a control solution contain-
ing only 10 mg/1 of chlorine in phosnhato-buf fered distilled
water was included in each experiment. T'ho four solutions of
test compound received nominal chlorine dn^r.rTps o^ 0, S, 10,
and 20 rug/1. Mixing was accomplished on a r-ul hiplo stirrinn
("jar test") apparatus, usino 1" < 3" stai-iTess steel naddlos
ro.tating at 80 rpm. Solution temperatures worn maintained at
25:°C - l°C.
Samples were withdrawn at 0.5, 1.0, 2.0, nr.c' .->.i hours ,-fter
chlorine addition and im.mediatel" analvr.cd ^or free and com-
bined residual chlorine. The orthotol i;:inc- (nr) and ort-.ho-
,r,
toliJine-arscnitc procedures, as d. . ascribe. -1 in Standard "r-thods ,
were followed in these determinations . Whore oh romatoo r c-'.^h i c
calibrations were- available, the sa^o'.es were also ana.lvzed
for the presence of the test, comnound. A sufficient quantitv
of sodium thiosulfate >.'as added, to eac'i si~r-!o ori^r to
chromatographic analysis to destroy the free chlorine resi-
dual. This step was taken to preclude the nossibilitv of anv
extraneous compound ch. lor i no renctiop.r, «.:uri nc exnosure to the
e].evated temoerature conditions rf the chromatooranhie column.
Tiiose compounds which had been fib served to react with chlor-
ine in the preliminary chloriiv.it ion nxpnrirc".t'-'; '-/ere sub-ic.cten
to further exardna.tion in a series oc detailed, tests. T.nc
experimental procedures were (.ssentin.n v the same as dcsor.ibed
above. A wider ranne of anp.lied ch lori ;ie riopnricjs (\\r> to 1.00
ng/1) was employed in these studies in an atter-it to ^urther
elucidate the. stoich j.ometrv of these re-lotions. I51t.ravio3.pt
absorption spoctropnotorne trie procedures '..TO used to nrovido
sup-piemen tal information on nnrent cor"r .'.".:. d. isnnT^oaranc^ and
reaction product formation.
- 20 -
-------
SKCTIOX VII
Pj:SbLTS OP PHASE I
Analytical Methods
On the basis of previous experience, chromatoqraphic techni-
ques, appeared to offer promise for the separation and quanti-
tative detection of the compounds selected for study. It was
envisioned that these procedures could also provide the means
for the identification of products of biological degradation
and/or chlorinotion.
Of the' fourteen chcm.i.caJ s initially selected, reliable chro-
matoqraphic techniques wore developed for seven. These .com-
pounds Wv-:rc amenable to chroma toy raphic analysis at low " (no/1}
concentrations using the Poropak Q column. Peaks wore well
separatF . from, the water response and observed to be symmetri-
cal, with little or no tailir.q. Reasonable response times
were obtained for these ricitc.ri5.lG, as summarized in Table 3.
Note' that thu relative retention times of: these species arc
sufficiently different, so as to allow the resolution of each
of the commoner.ts in a complex mixture.
TABLE 3
Ci.j-.criATOC.RADilC ANALYSIS OF TEGT COMPOr.XDP
RiiTnKTiOM DATA
Column: 6' 1/8" Poropak Q 100/120 Mc-i-h
Compound
?''.o t h a no 1
i.'sopropano.I.
T-Butanol
Acetone
benzene
7'oluop.e
Lt'nylben.-:ene
Carrier Flow
/.,, 1 -. ; r N
v . i i. . ;..!/
35
35
35
35
3 5
35
20
Colunv.i
Tcru'orature
'(°C)
120
ISO
150
1 5 0
160
190
225
Hctenti on
Ti.no
. (mi n )
2 . s
5.5
9.5
4.0
11.25
10 . 0
7.0
AF; a re.'uilt o!. tiie lack rf suitable cas chj'o;"?.Vrv-:raph.i.c r\>-
ti.ucls i-'or th.o ar.aly^is of certain of the test cor.pru.--.dr-, ul-
traviolet absorption spact.rophqtor-.etric procedures vor-. a?ro
evai'jatci.1 durinn the r};a?o I c.;t\:c.ias. The result.1; of ore'; im-
inary tests confirmed the sui tsbi J:! t.y of UV analysis for v.he
Jetcrr.iinntions of several of th..-: -ost compounds, includinc
-------
'phenol,, m-cresol, nitrobenzene, aniline,.and hydronuinone.
Diraothylamine v;as found to be inactive in the \J\' region.
The absorption characteristics of the compounds tested are
summarized in Table 4. Subsequent experiments also revealed
the utility of UV analysis for chlorinated product identifi-
cation and monitoring. These studies will be detailed in a
later section of the reporc.
TABLE 4
ULTRAVIOLET ABSORPTION CHARACTERISTICS OF TEST COMPOUNDS
Compound ,\ (nm) ,\ ' (nm) \ (nm)
; _i .-! 3
Phenol 207 235 288
m-Cresol 222 271 277
Hydroquinone 193 220 283
Aniline 199 230 280
Nitrobenzene 195 . 214 270
4chloro3-methylphenol* a-200 279 287
2,4,6-trichlorophenol* -v214 245 312
2,4 ,6-trichlcroaniline* %210 242 304
p-bonzoquinonc* 245
*Studied in Phar.es II and III
Biological Degradation Studies
The purpose of this phase of the investigation was. to iden-
tify the test compounds or their degradation byproducts which
could react with chlorine under condition." that occur in mun-
icipal waste treatment. .Two conditions of treatment must be
identified; first the acclimated condition, when: the? test
substrate is in the system continuously or for a sufficient
period for acclimatization of the !. iclooical system to occur,
and second, the unacclimated, or partially acclimated system.
The unacclimated condition is the result of a njn-rcnular dis-
charge of a particular chemical to the treatment plant. Both
conditions regularly occur in practice.
Acclimated Systems
The setup of the physical systems has previously been des-
cribed. The control systems received a syntnetic "spv:a:jo"
with a COD strength of 200 mg/1. Each, of the test streams
were composed of a mixture containing 100 mc/1 COD of synthe-
tic sewage and 100 mg/1 as COD (calculated basis) of the test
substrate. The effluent from each test system was examined
to identify the' component or its degradation products in
effluent.
- 22 -
-------
Except in the case where the test substrate or its known de-
gradation products are directly measurcabie, a significant
determination problem results. This problem can, in most
cases, be surmounted by considering supplementary data on
COD removal, and degradation studies on the particular com-
pound. Employing a combination of the analytical information,
it is possible to examine each of the test compounds. Table
5 summarizes the studv results.
. I TABLE 5
SUMMARY OF BIOLOGICAL DEGRADATION STUDIES
Compound Results
T , Complete losr; of primary substrate (oas chroma -
Isopropciiiol . , , T, . _.,,-'. , ,. ,-
tccjrapliy) . Idcntiflaolc intermediate acetone.
,, . , Complete loss of primary substrate (oas chrcma-
tocjraphy) . Mo identifiable degradation product.
'm-Crcsol Biological oxidation of substrate.
Phenol Biological oxidation of substrate.
Complete loss of primary substrate (gas chroma-
tography). No identifiable degradation product.
Benzene Removal by stripping and biooxidation.
Toluene . Removal by stripping and biooxidation.
Ethylbenzene Removal by stripping avid biooxidation.
Presumptive evidence of complete loss of pri-
Benzoic Acid mary substrate with no degradation product for-
mation. biological oxidation.
II" Irortr ' o-«e Possible substrata persistence or degradation
''' ' product formation.
Coriplcte loss of primary substrate (gas chroma-
Nitrobenzene tograph). Possible formation of unidentified
degradation product.
Presumptive evidence for some primary .r,uLstrate
Diroethyj.amine survival. Armenia is a produce of biological
oxidation.
, r. , , Pc.rtial survival of v-rinarv substrate, dooraJa-
i-butauol . . . , , .-
tion product torniation unknov:n.
-. 1 % :''ossib.lf survival of ;-rir.-.ary substrate suspec-
ted. I.onradation product f or nation suspected.
Figure 2 prosc-ntr, batch study data illustrating t;:e disappear-
ance of isoprapanol \-:ith a corrc'-st end i ng increase in acetone,
-------
-\ --A\'OL A!K ST9 JP!f-."j DATA-.
NG- HO'JRS
FIGURE 2
BIOLOGICAL DEGRADATION OF ISO^ROPANOL
-------
as ' determined-by chrbmatographic -analysis. A complete stoich-
iometiic evaluation of the completeness of reaction was diffi-
cult because of acetone loss by stripping. No evidence of any
other product formation was uncovered. Acetone itself was
eventually lest from the system at. longer aeration times (see
below}'.
Methanol disappearance, as measured by gas chromatography, and
COD disappearance data indicated that msthanol is completely
degraded without the production of an intermediate product.
Comparison studies indicated that strapping is not a major
factor in removal, as shown in Figure 3.
Both phenol and in-cresol could be tracked by gas chromato-
graphy and COD analysis. A typical gas chromatcgraphic output
for m.-cresol is presented in Figure 4. In another cxnorimFnt,
the phenol concentration was reduced from 33.7 mg/1 to 4.7
mg/1 after one hour. An analysis of residual COD d:ta, as
shown in Table 5, indicated that no intermediate products were
present in detectable quantity.
Acetone was monitored by' gas chromatography and showed cor-
plete disappearance from the test units. COD date. U'efer to
Table 6) indicated no accumulation of an additional product.
Examination of the removal pathway indicated that a signifi-
cant portion of the acetone was removed by stripping, a? 'evi-
dent from Figure 4. Approximately 50?; of the renewal was ac-
complished by biological oxidation.
T'.BLE 6
SUMMARY OF COD DATA FROM BIOLOGICAL DEGRADATION STUDIES
Compound
Acetone
"sopropanol
;a-C'resol
.Methanol
The no 1
Ben ;:er.e
Eti.ylL.eni'.ene
Toluene
Ben zoic Acid
Hvdrocuip.one
Nitrobenzene
Dime thylanine
T-Butano].
Aniline
COI.VKCL .
! Includes 100
*"'..';. Icula ted i:
Theoretical Total
TOO 7'- -Irlodt
V* Wi-^ '. * -1 *«-l V, V_» 1
200
200
200
200
200
200
200
200
200
200
200
200
200
200
-200 .
.mg/1 synthetic ncwag
rom Table 2.
I n i t i a 1
COD Recoverable
Bv Test *
'
151
175
150
193
1 9 5
104
101
1 0 4
200
1 9 7
106
100
164
152
200
e (as COi:) .
Residual
COD
(mg/1)
n t- ?'- '- 1-
Cl ... tL -1 i . .1
AV crane
-11.6
50.1 .
4 ': . 7
-19.3
4 j. . 8
4 8 . 5
5 i? . 2
45.3
2- F . }
3?.0
~ ? :'
5 :- . 0
6 9 . 9
68 . 5
5 f. t -,
-------
60
_J
V.
o
o
t-
2°
TIM?-HOURS
_0 _ __
FIG'JRE 3
BIOLOGICAL DEGRADATION OF ACETONE AND ME7HANOL
-------
\:
FIGURE 4
CHROMATOGRAMS ILLUSTRATING M-CRESOL BIODEGRADATION
-------
The removals of benzer.e, toluene, and etnyibcn/.ene. were moni-
.tored by gas chromatography. Studies indicated that they
'.-.ere, to a very significant degree, removed from the system
by stripping. if .any biological 'degradation products v:ero
formed by a small portion of these materials that were ac-
tually oxidizc-d, th-zy were bclov: detection limits.
A primary substrate method for evaluating benzoic acid pre-
sence v.'as not available. COD date, (refer to Table 6) indi-
cated complete oxidation of the suostrate with no app£ircnt
degradation products. For supporting evidence, a sciir.ple was
'studied for the regularity of oxygen use with both acclimated
and unacclinated seed. The results of this study are shov;n
in Figure 5. The regularity of both curves supports the con-
tention that the reaction is complete, with no significant
intermediates being formed. Although the evidence is not
totally conclusive, there is sufficient presumptive evidence
to infer the improbability of degradation product formation.
The lack of a totally satisfactory method of analysis made
determination of hyurcquinor.e in low concentrations imprac-
tical. The substrate was substantially degraded, as shov/r.
by the residual COD data in Table G. However, indirect
oxygen utilization data suggested imcomplete oxidation, or
possible degradation product formation. Possible products
v:ere not identified experimentally.
Cas chromatographic studies indicated that nitrobenzene dis-
appeared from solution during the study period. Analysis by
ultraviolet absorption indicated that removal by air strip-
ping was insignificant. CO!; and oxygen use data suggested
the formation of .a degradation product of the reaction. Th^
product could not be identified ext:ori.~:er.ta] ly in this study.
The product concentration was in thr order of 10%-lSx, of the
original nitrobenzene as CO'J.
Although no satisfactory method of analysis was available
for low concentrations of dinet'r.ylarninc, there is presump-
tive evidence that some pri.nary substrata persists for n
long period during biological treatment. 7-.nnor.ia v.-as found
as a degradation product of oxidation..
T-butanol degrades veiy slov:iy snd is detectnbl-'j even after
exhaustive periods of biological oxidation. '.To degradation
product was identified, but consiclori.no the r,mount of the
t-butanol present, it cannot be positively concluded that a
product does not exist. Typical degradation data arc shown
in Figure 6.
'.'o suitable analytical method was available fcr low concentra-
tions of aniline. Based on COD data, (Ta^lc 6) it may be con-
cluded that 'either aniline or a degradation product remains,
even after substantial tir.c. The .nnalyticil evidence is in-
sufficient to dr.~w a valid conclusion. However, indirect oxy-
gen utilization studies suggested the persistence of aniline
ti.rough biolcgioal systems, particularly .in t'rf- nrescr.ce of
mixed substrate.-.
- 28 -
-------
500
! « 400
FIGURE z
BOD PROGRESSiC.v C^ BEiVZOIC ACID
-------
VOLATILIZATION OF T-BUTANOL
BIOLOGICAL TREATMENT Or T-3UTANOL
-------
Unacclimatod Systems
In an unacciirnated system, the discharge n>ay contain any of
the products found in the acclimated system, as wall as the
original test substrate. The concentration and distribution
of products v/ill depend on the actual conditions of discharge
and treatment.
Discussion of Results
The results of the biological degradation studies indicated
possible degradation product formation in rcclinated systems,
for the follov.-ing test substrates:
aniline
hyclroou: -tone
n i t roh/en zone
t-butsnol
isoprepanel
diir.e'.-hyiair.inc
Airononia was determined to be the product of dimethylamine de-
gradation, and acetone v:as identified as an intermediate in
isopropanol breakdov;n. Due to the liir.itatioriR of the available
analytical netnods, de-gradation products of the remaining ma-
terials could not be confirmed or identified. As a consequence,
it was decided to concentrate efforts on the determination of
problems associated v/ih chlorination of the original test
chemicals. This approach has direct application t<. "spill"
situations, in which a substrate: passes through an unacclimated
biological system, without degradation. Such occurrences are
commonplace in industrial wastev.-acer treatment practice.
Chi orination Experiments
The first series of chlcf-ination experiments v.-as designed to
determine which of the selectee: chemicals were capable of re-
acting with frue chlorine under conditions commonly encountered
in conventional treatn'.-nt plants. A portion of these prelimin-
ary investigations v;as devoted to the evaluation of the preci-
sion of the residual chlorine test and the stability of chlorine
in test solutions. 7-. .precision' of 5\ (relative standard devia-
tion) was .obtained fron a statistical t'reatr'cnt of thirteen rep-
licate chlorine residual determinations on control solutions
having nominal chlorine concentrations of 10 r.ig/1. In accord-
ance with this observation, a froi': chlorine demand of 101- of
the applied dosage v;a£j the- criterion used to dcterninc any sig-
nificant compoundchlorine reaction.
The results of the- -first sor:.es of cc:"rou.-.d chl or i nation exper-
iments are given in Tahlc 7, >::.ich lists the chlorine demand as
a function of applied chlorine coaice nr.:l contact timr. for each
of the tost chemical.-,. The cata indicate.- than, under t.h° test
conditions, no significant c: Icrir.e ':'-:".ir.d :::.:: c.-xc-rci sod by the
fol lov.'inci cor.oouncs:
-------
TADLIJ 7
;[.TS or P^;:LI:-II;IA?Y CTLOPINATION
Nominal
Cl.lotine Demand (OTA) At Indicated
i_f_i;iii-/uui:_i i..;y/j.;
;-U-. thanol, 8
Isopropanol , 10
T-Duta.no 1, 10
Acetone, 10
Acetone, 20
Scnzer.e, 10
Toluene, 10
n'-'-.ylLer.zene.-, 17
iienzoic Acid, 10
P'/.er.ol , 13 . 'J
n.-Cre/sol, 20
Hycl-.:c::j:.nc.:-:c.-, 10
Aniline, 10
Uinothylarr.i -.'-., 2
;.'itroLer.7.ciic , LO
sr L; L;. '.:.
f'r 1 p .
>16.
.^s .
M6.
7 '
' 1 C .
- o .
KJ .
:'. ."on : re
^..
l>
3
fj
G
j.
6
2
2
2
0
1
7
1
0
2
r .
0
0
0
0
1
0
1
f 1
0
0
r
UK U
J.
-0
-0
-1
+ 0
-0
-0
*0
-0
-fO
-0
-0
-J-0
-0
+ 0
+ 0
-i-O
> 8
11
-G
+ 0
tor
ct(_ U
.0
. 6
.7
.1
.8
0
f-
0
0
t
0
.3
.3
. 3
0
.2
0
0
. 2
,2
-
-
.0
. 1
. C
-
'-
(\
, 4
:.-os
J. J-ilitJ f
J.
-0
-1
-0
-0
-0
-0
-0
-0
-0
-0
+ 0
-0
-0
+ 0
-0
+ 0
-0
..-0
+ 0
+ 0
-0
+ 0
+ Q
40
idual
:iuu
.0
.1
.2
.2
.4
. 9
0
0
.4
.8
. 1
.2
.2
.3
.2
0
.2
0
.2
-
-
-
-
-
. 2
0
t on
r :;
24
-0
-1
+ 0
+ 0
+ 0
+ 0
+ 0
-0
-0
-0
- v
-0
-0
+ 0
+ 0
-(-0
+ 0
lie. i n t
.0
. 5
0
.0
. 3
.6
.8
.-I
0
. 6
_
:
.3
.7
.4
0
0
0
0
-i
-
-
-
-
-
.3
o
or-t
-------
jcotoiir-
to] .:-<««
o*~-hv locri /.r*n^ *
"li^r^ijor zono
Tnose corr.o'jr.'i.s r.nrko:! v;ith on ast-'ris;' (*) vo_o ronitorn-I bv
ess c/: r'..'~a"or;r anh-v ana cxh. j b.i to'.; no s ian j fic'int not corr)ounri
Io.v3 (corr,o ,;n-2 control rinu?; trvst} rirrir.cr the. ^x~er.i.r-2r.'-.s
."-oroovo;f, no rocc-n'lr. / peai-'.s '-/ore obssrvfc in onv of tr G chro-
ratovr-jrs to indicate the ox-is tenco c~ anv r '':'.:': ion pro.'l^.rts .
T ; shcul'": !";'- ootO': that be:.x<".'T.r:, "zoluer.o, an:" ';thv Iher. z---:1. <> '-'or
rj-..i'"]v il-',-"? t fror both to1;^ -^r:',: control =;oi. r. ~ ion1; r";urir." ri:or:::cr:'.j , -'" 3h'7-T. in Fiour^ 7.
71" ov.i'V.rt fror Tobl0 7, chlorine vras oh^orvo''1 to rp-ic'; ;.! tM
oh(':nol:, :- c ''": ^ '; 1 , h"-'lrooi!ir.or;ri. -in i? ino, a~:.'l ^irr- f-'-;'/l.p.ri-, -.
P:,C:;:I! ''..':.' r~/;.}, r-Cro-o] (?.'i r.-:/l) f.n-f' .i-.i'.iro (1.0 ~o/l)
cornlot '!' ccD'-r'-irc'1 cill o ~ the arv. lie'" chlorir.'? uo t"- tho
hinho^t :o';'?':-i (lr. .2 r.c/I) v; i '.-.'-. i. p. i. contact ri---! or 0 . '. hours.
iiy^j'cquinor.',-- {I") :"'-/!) anc; c":-.-r: -hvlari. n-- (2^ r<-/l ) cor-l.-Tiolv
ror'Ovo'J ?. 1 1 of the. -v-Til ic-"' chlorine at ~h. -3 t'-to ]'". or c1oc. .T'^cr;
{'.0, ?.l rv/1; -if tor a 0.5 :/o-.;r conta -?t tiro.
ji-i'.'in^ i 'J'': r. '-. i f i 'vii tho^o corr;r;"jn':s v/h- i. rh 'iG:~r-. cnoab.le or r^:?.c-
tir:'.' v:j. -.h '_'". lor inn , a roro u'^tGilc:-? G:-:r,ori~-ontal rrocr^r1 '-.'as
:1 -. itiatsr: ~-j r.rovic"o .-juonlcr'or.tal. infr.rr":t ion rocr-:r'"i"c t.-toi. r
ci' ' " r i ;": :' ~ .' ".-" 'jror<"-rt ios . "'h.or.o st'/'lios v;crc- n^0; i':ne"/ to: (1)
ru'-t.'ic-r "'fin': _h'- chlorjno ':nr;,n-:^ o-~ cich of v.:," choricnls,
-'ct ion , an^" (3)
('!') I'Stz^lL**.':* tihr- contact tirra
i -.:c.-j. '. i. :' , '.''.-' ro r.r-n ^ in lo , "ho f.or.-r;t inr: o^ sr.o~; fie croquets.
7;.c- c-Tf; -;r L" r-r.-n 1 cor.rii ti^ns o:' -hj"; sorifs o~ jxnoriron':- v/orr:
/ o nor z 1 ;.'' s;.-il.--;r 10 chose pr^v: '"jslv doscribr--: (T:.;: '/.^t 2"°C,
ofc.; , h'.'/r----"r , :i v;i'1=!r ::ir-.c:: of corsonr. -1 an.r" aor.'. ic-'" chlorine-
P.-iCMio] V/.-JT oxnr i nfr'i at c")"ce.-:*,r-:-ti^n? of n --Tf: 2""' ~'c/l, --. -h
.'.:.:. J i o;: 7h ' :, r '! r. r. ".o ^ac;os o" 20, o1" fjnr 1''0 r:-/}. The=;p !?:'.:
nr-'.- o ro .s 7 r. ; o ' in Tonic 3 anc J'icuro 3. It is r.r. .tre :t thnt
'-;':' rio 1 ^ .".'.' '_ r*'-* -.o o " ~i rol nt i*."ol"f rr.nf " rc;r:C". i '"-r '"' th chl^rir.0
i 2° -'
;tt;cr '>."'>"., . ~ "' "'' cal-:;v. l-^t-.-'" ".h^1- :
I">ri TV.- .:::: ": of .' . 3 ": re: I'l^-'rv oh^n^:
/
i',"*_ ;iir"': . '.* ! ^ i " is c**i'.ivr] C-T*". "-'i '* . ^ r'
o- nh^no?. cor-;"1. Pto TV
"r I' .-"'1 or rh^nol,
.'."". '. xr>rci~c>" -
-------
tjj
<_>
2T
o
O
!JJ
LJ
20
16 -
0
:_/:_ (COM-'-OL)
O-iO VGCL. -L
VOLATiLIZA^'CX OF E'HVL3E\ZEKZ DURiNG
-------
-------
r.olc.of phcr.ol. .The consuration o" c'llorinc :iv r>ne".ol i.~,
?.l.~o sho--;a in -Fiaurft 9, in which t'vr? 'V.-n hn"<-> bo^n exorossoo
i:-. tarrs of nolar ratio." (r.-.ol chlorine nor rr~>1 oaonol) .
TA3L.r: S
C!iLORi:TATi.;-: f'"!i:op2.7
0 : 2 0 - 2 . 7
17. ° 3^.° -'-.1
? 2 4 2 ^ A
o. 2ir'"> ':-r-
~> ^ r. 1 r- . K
:' rjf 7..-1
3° P'1 ^ 1
1 ' ~ .', ? ~ ~ '
-! . " -:' =5 . "> ." . n
fi r)a -? . "-
0 ~r~ ~3. "'
0 FS" ^3.3
12 ~ " " . "
o - a 7 ' fi ~
» ' .
rj ^. T ^ o -, .*-j ^
0 *>1^"! '- "' . C
A-~\- ir'-rl-^v f- -,ror:uctr,
ch lorv'.f ''-.h ::"''.onol, v:sin«
:ay. 7n\. In th -~ n X" or ipon t , a
ojr.ti.~iri c'intaini.nn 25 r\c. /1 of p1-! c"."1 3. a-.'1 2" r~. '1 o~ c:-.lorine
:.':: :>:\v-arpf:. '."'lii.^; soloct i .">:-, oc co-iror.': r.it .!"".." :at; race to
*"."! '-^ ar. oMcms o ^ '^h cr/^l , ro ?.?.*". i "o t"> c%: Virf. ". o , i ~ir/'Ioj" to
_L . j ': .:c the- posr. ih.i li t'/ of ciiract c!i lor? r.c c:-:'.r"~" i°r. or tho
!io;-r> ". i - -s tr'.ict urr- . A:"'-~r ;; c~>ntac" tr ^ o' ~." -."<:.:r'r , a sa~-
:V. i Iv; i M . ."4 ;n t j ci -:; to ": , :i ^ c'~l~r;.:-i r^=; i ";~". ~:.= \otootod.
. ".i o r ~. '~c"v"ai~.o>~: rap o">i a~. r.c1-:: on
7',]..r>...n.;r.i,.,.n:..
-------
o .-;. r
6 3 10 12
CI'LO;?iNE APPLIED
(!.! l.iOL r. I. -. / M fvlOL COMPOUND)
14
MGURE 9
MOLAR CHLORINE UPTAKE GY TEST COMPOUNDS
-------
" '.':! (" E _ ,-T' V T. 0X' T S
FIGURE 10
CHROMATOG'RAMS OF PHENOL
CHLOR,'N!Ai"ED PHENOL SOLUTIONS
-------
These wore subsequently identified, on the basis of relative
retention data, as o-chloropher.ol (1), 2,6 dichlorophenol
(3), 2,4 dichlorophenol (4), 2,4,6 trichlorcphenol (5), and
p-chlorophenol (6).
The results of similar experiments, using m-cresol as the test
corn-pound are shown in Table 9 and Figures 11 and 12. Tha re-
action of chlorine with m-cresol also proceeded quite rapidly
in the first 15 minutes, and was essentially complete after
two hours contact time. In the case of m-crcsol (10 and 20
mg/1), the application of 50 and 100 mg/1 of chlorine produced
a free chlorine residual after two hours. The maximum observed
chlorine uptake (after 2 hours) was computed to be 3.84 nc Cl?/
mg m-cresol (5.9 moles Clp/mole m-cresol), corresponding to
the solution which initially contained 10 mg/1 of m-cresol and
100 mg/1 of chlorine. The consumption of chlorine by m-cresol
is also shown in Figure 9, and indicates that m-cresol exer-
cises a lower chlorine demand than phenol, on a molar basis.
TABLE 9
CHLORINATION OF m-CRESOL
m-Cresol Applied Contact
Concentration Chlorine Tine
(mg/1) (mg/1) (hr)
0
10 20 J
2
0
10 so ;
2
0
1C 100 °
2
0
20 50 °
2
0
20 ' 100 °
>
.25
.5
.0
.0
.25
.5
.0
.0 .
.25
. 5
.0 .
.0
.25
. 5
.0
.0
.25
.5
.0
.0
(OTA)
Chlorine
Residua
(mg/1)
3.
1.
0.
0.
30.
30.
28.
17.
81.
77.
61.
61.
16.
11.
8.
S.
61.
58.
56'.
46.
3
5
5
2
8
8
3
0
i-i
0
6
6
3
1
0
0
6
2
6
0
Net Chlor
Demand
inc
1 mg/1 mnol C12
Tino! m-Cresol
16.
18.
19.
19.
19.
19.
21.
33.
18.
23
38.
38.
33.
38.
42
42
38..
41.
43.
r .1
-J *!
7
5
T;
8
2
2
7
0
6
4
4
7
9
4
8
4
0
?
2
i
3
5
?
3
5
>
3
C
5
2
3
3
3
2
3
4
. D
.3
.0
.0
.9
.9
. 3
.0
. c
. 5
_ 9
.9
.6
.0
m 2
.2
.9
_ 2
. 3
. 1
39 -
-------
-------
T~l
I i
i ,
J i/
FIGURE 12
CHRC'.'ATQGRAMS Or M-CRESCL
;H_C-;SATE:> J/.-CRCSOL SOLUTIONS
-------
Chroma tograrns obtained for m-cresol and a chlorinated solution
of m-cresol arc' shown in Figure 12. Several additional peaks
are evident for tlK: latter sample, reflecting the formation ,
of a -complex mixture of products. None of these species could
be positively identified, due to the lack of commercially
available chlorino-substituted cresols. However, it is prob-
able i that the products constitute a mixture of chloro-n-cresols
and oxidized forms.
The reaction of chlorine with hyclroquinone was studied next,
using compound concentrations of 10 and 25 ng/1, with nominal
applied chlorine dosages of 10, 20, 25, and 50 rug/1. The
data for this experiment are presented in Table 1C, and Figure
13. As in the case of phenol and m-cresol, a rapid initial
reaction v/as observed, followed by a declining rate of chlor-
ine uptake. The reaction appeared to be complete after tvo
hours. A maximum chlorine demand of 2.37 mcj Cl/ng hydroqui-
none (3.9 moJ es/mole) was observed under the test conditions.
(The molar consumption of chlorine is plotted in Fiqure 9,
and shov/s a lower chlorine deraand by hydrcquinone , relative
to phenol.)
TABLE 10
CHLORINATION OF HYDROQUINONE
Hydroquinone Applied Contact
Concentration Chlorine Time
(mg/1) (mg/1) (hr)
10
10
10
10
0
f\
10.4 . ^
2
0
?0.8 J
2
0
1C ^
1.
2
0
52 J
2
0
Q
1
2
.25
s
.0
.0
.25
.5
.0
.0
.25
. 5
.0
.0
.25
. 5
.0
.0
.25
.5
.0
.0
(OTA)
Chlorine
Residual
/ /i ma
(mg/1)
6
4
'2
1
9
10
5
4
30
32
30
28
9
7
3
1
0
n
0
0
,
.
.
»
t
*
.
%
,
B
B
1
75
75
8
8
0
5
5
3 .
7
0
3
^
5
5 .
0
>10
>" 1 0
>10
i?-°
1'!
1C
18
19
Id
16
20
21
2.1
19
22
23
4"
44
48
51
Ket Chlorine
Demand
/I OT
.4
4
.4 '
.4
.7
.05
.05
.0
. .i
.0
.5
.5
. 2
.3
.0
~J
, L
-,
.5
.0. '
nol C19
ll H-'
>1.
^~ -i
^"1 .
> I .
2.
2 .
2.
2.
" .
2 .
o .
3 .
3 .
3.
3 .
^. .
2.
3 .
o
-<
e,
c,
6
6
3
5
r-
^i
5
2
3
3
0
7
6
3
0
2
- 42 -
-------
CO
LJ
:r
10 M? HrDrtoo'jiMo;.?: /L
L - r. > ."-./ "*
FIGURE 13
REACTION OF CHLORiXE WiTH KYDROC'JiNONE
-------
In the absence cf on- ';;-pJ -'"-'-'.-- chrorvstociraphic procedure, the
use o;i ultraviolet ---.i.-or:.-..or. :;pocrr or,ho tone try for reaction
predict identification vo:: oxoj.o-red . Oaring the previous
studio.,, it ..-as no tod t.-.ui tho addition of chlorine to hydro-
:;uirono occasio.-.-':.My rc.zui-.od in the production of orange
colored solutions. This ho-ravior V.-.TS particularly evident at
the highc-r reactar.t cor contritions. Such phenomena are not
unconrr-on in aquc-cu:; :.;olv> v1 on:: of aromatic organic chemicals,
and gone-rally arc- tr.o ro^uii of oartiaj. oxidation of the ring
str1. cturo. It v;as thus r,o:-. t > <. vtod that hydroqui none reacts
to form p-benzoquinone as
0
ii
I;
0
To test this hypotoo^is, a separate chlorination experiment
?.s conducted. (JI: c -.-:.; ol-.:: :,; octra \.-er-: first obtained on
bufforod (pH 7.4) ."CJ.utlor.:i of both h'/droquinone (10 and 20
r.g/1), and p-benzc-juirc-n'.: ('_, 2, and 5 r,-.;/l) , as shown in Fig-
uro 14. ::ote that hydro'-j i nono ox::i bit;; absorption maxima
at "-22G n~! and '-'/.'<.''. c-.:'*, v.'horoa': T-.--': <-;r.7.ov;uinonc absorbs maxi-
r.ally at '-245 n::.. The L"7 n:.:-.-^ r:..r i on chnract-.eristics of each
cf the pure compounds are tnuc. ::ufficiently different to a.l-
icv: their identification in a ::.: ::ed cr.nplc.
.-. solution containir.',: 10 r'.g/l of hydrc-:uir.or.e aiid 20 mg/1
ci". Jorine v/as next nro- arod. '.his c:orresponds to an applied
chlorine to hydroquinone -o:ar ratio of approximately 3:1,
in correspondence- to the previously observed chlorine demand.
-.& in previous studies, the p?{ v.-as nainrair.cd at 7.-1, using
tho phosphate buff or. 7 f'.or a conta<;t tirr.c of 10 sninutes, a
-2.--.pie v.-a.s v:ithdrav;n for ."." ^n-vlysis. The resulting spectrum
Ls also shown in Figure 1 , i:.d demonstrates the rapid forma-
tion of p-ber.2ocuinc.no, r.-r: : r.'' \ catod by the appearance of an
absorption r.axinu.~. at "-il^D r.". Correspond ingly, the peak at
2Z.?. nr-, i.-; observe'; oo d::"ir.i:.:-, rofIr-ctir.c; a substantial de-
crease in the initial hydro-;uinonc concentration. The peak
:.t '-245 r.~ ?o-honxo';u: nono j v.-ac. found to gradually diminish
-.urine: subsequent ~o-u.'-;urc::-on-;.i over tv.^.'.-.ty-four hours. Cor-
ro'-;pondin^ly, a br:--'id "nhoul-j-i-r" in tho 265 - 295 nm region
;.:volcp;:d over chi- :;:.i." o. ronou. Tho ;.: ^-cies associated vith
.:.-.-> absorption ::--.:- not j-:-jnti f io;;.
..n: line v.'as also ro-oxa.T.ine'j in the .scries of detailed chlor-
in::tion oxr,c-ri~ents. .-'-. n:. 1 ic-'.i chlorine.- dosages of 18, 45, and
-------
240 290 l'-r.
WAVELENGTH-KixNO.'v'ETE^S
FIGURE; i
-------
90 no/I v;orc added to solutions containing 10 and 20 ing/1 of
anilirio. The data derived from these tests are -jiven in Table
II and I^igure 15.
TABLE 11
CHLORINATION OF ANILINE
Aniline Applied Contact
Concentration Chlorine Time
. (ir.g/1) (mg/1) (hr)
j
0.
0.
10 18 1.
2.
3.
0.
0.
' , 10 45 1.
' 2.
3.
0.
0.
10 90 1.
2.
3.
0.
C.
20 45 1.
2.
3.
0.
0.
20 90 1.
2.
3.
25
5
0
0
0
25
5
0
0
0
25
5
0
0
0
25
5
0
0
0
25
5
0
0
0
(OTA)
Chlorine
Residua!
(me,/! )
1.
0.
0.
0.
0.
27.
20
14
6
4
58
52
43
34
28
4 .
0.
0.
0.
0.
40
30
.10
3.
2.
75
4
2
2
2
5
0
8
G
2
2
6
G
:."et Chlorine
2 Demand
1 mg/1
.pv
u"
16.25
17.6
>17.8
>17.S
717.3
17.5
25
31
39
41
32
38
47
56
62
41
44.2
-44.4
744.3
~44.8
50
GO
80
86.4
87.4
mmol C
.\ "1 A Tl T
iW J_ f \ i 1 J.
2.
2.
>2.
72.
*~ "
2.
3 .
4.
5.
5.
4.
5.
6.
7.
8.
2 .
2.
>2 .
72 .
I2'
3.
3.
5 .
5 .
5.
L2
-i ' _ -.
J. X 1 1 1;
1
3
3
3
3
j.
3
1
1
4
2
0
2
3
1
7
9
9
9
9
3
9
3
7
7
As previously observed for hydroouinone, the appearance of
orange colored solutions was evident at'the higher applied
chlorine levels (50 an<_; 100 mg/1). It may be noted from the
figure uha~ aniline also initially undergoes a rapid reaction
with chlorine, follov.ed by a declining rahe of chlorine up-
take. Chlorine consumption war. observed to continue over
contact, tir.es in excess of two hours. The ruixi-^n chlorine
uptake at three hours was calculated to be 6.2
line (y.l roles/mole).
Cl2/mg
- 46 -
-------
"
FIGURE i5
REACTION OF CHLORINE WITH ANILINE
-------
In a separate experiment, an attempt was made to gather m.ore
information pertaining to the nature of the reaction pror.urts
of chlorine and aniline. In the absence of reliable chrema-
tographic techniques, ultraviolet spectrophotometric mothcdr
v.'erc employed for these tests. Solutions were prepared, in
accordance with previously described procedures, certainino
10 mg/1 of aniline, to which chlorine dosages of 5, 10, and
20 mg/1 were added. Samples were withdrawn at varyinr inter-
vals of time for L'V spectral scans. Kecordod spectra fro:-:
this sequence of measurements are shown in ri:;urcrf 16 and ] 7 .
Figure 16 illustrates the effect of chlorine dosaoo on tho
ultraviolet spectra of chlorine-contacted aniline solution::-.
The scans shown were taken on samples after a contact tire cf
one hour. Tho spectrum indicated by a dashed lir.o in I'igurt.
16 is that of a id ng/1 aniline solution, to which no ch.lorino
had been added, and is included for comparison. Aniline
alone exhibits fairly well resolved absorption maxima cen-
tered at 2"i(i nm and 281 nm, with no significant absorption
above 31C n.v,. The addition of 5 mg/1 of chlorine has tho-
effect of shifting the peak ahsorbance to about 233 r.m in a
broad band which extends co 330 nm, with evidence of unre-
solved inflections, and a shoulder in tho region near 2S5 r.m.
7\ dcsagc of 10 mg/1 of chlorine produces a further -rhict -ir.
the absorption pea.-: to about 252 nr,. '. broad band is r-.-->.in
evident which extends to 3GO nm and rota ins the shoulder
around 285 nm. The highest chlorine addition. (2J r.r/] } was
observed to criur-o 2 further :;hift of ~he most prcmironv. poah
to about 267 nm. The broadness of the absorption bar.d : r
aiso increased, extcndir.n to about 370 :\~, ard shcwi.".o sovo-
ral unresolved inflections between 220 and 270 nm.
Ultraviolet absorption spectra wore a'Jr.o veoord.ed at varyir."
time increments for the 10 mg/1 atulino r 10 mr/1 chlorir.o
solution. Those results aro i 1 lustravo '. in Pio-viv 17.
The first scan was obtained after 10 minutes contact, tiro a-.d
shows a si:i-ft ::: the aiscrption peak, producing: two porr'y-
rcsol\'od ;.'axi:";a :i t about 253 r.m ai'.d 2J.',"1 nr, wi t :. a r.'"";-". d.-r
cer.tered arov.nd 286 n.r;. Aftur a contacv time of o:-c h.ov.r,
there is no evidence of tlie dual, poahs :>.L ~52 nr ar.; l.i' :.r ,
but a sinclo bread a; sorption peak now a; ; o ;r.- a
:;oto also Li-nt the a! sorbancc of ti'c ah: 'Oder ha
/ final sca;j, U'.":-':-. at t'-.'o hours, siicws a furtii
the bread band maximum to about. 2-if nr'.. Those
cor.sisten1- v:ith ti^e data developed in i he chlcr
studies, and confirm the :?rc;-:rnssivc naturo cf ~ .c
chlorine reaction over contact timer- in excess of two h v. r r-.
Speczrn v.ere obtained on dilute aoueous sclution:- of sc-vora.:
chlcrine-subctituted analinos. The absorption maxima cf i-tr'
compound are as foilov/s:
-------
00 -
"o'1 C-'./WV?
I O
OC
CO !-
10 mn/l A\'1L!\'E »
10 i'":/I CH..CRI','!-;
: -0.'> C'j-; v.C"
F--~'-TRA OF CHLOrviVir CONiTAC
'.'--"C' 0- CriLOSi.Vi DOSAGE
-------
oo p ,:...T
r j' ' 0 ! '
\
'
2<0 290 340
WAVELENGTH-NANOMETERS
FIGURE 17
ULTRAVIOLET A3S03PT',ON SCECTRA OF CHLORINE CO\ITACTEC
AiSTJNE SOLUTION'S: EFFECT OF CO\|T;CT T'ME
-------
aniline
o- ch lore ar. i 1 i ne
p-chloroaniline
2,4-dichloroaniline
2,6-dichloroaniline
2,4 , G-trichloroani 1 ir.c
220
222
22S.
2-;o
228-72C
' =s
A in a >:
B-hand (r.n)
*).' 7
231
286,
293
04
It is evident that chlorine substituticn on the aniline struc-
ture promotes a bachochrcmic shift in the wavelengths of maxi-
mum absorption. This effect is enhanced with, increasing sub-
stitution and is more pronounced with para-substitution, than
with ortho-substitution. A comparison of Figure lf> with the
data given above reveals that none of the spectra obtained on
the pure chloroanilines can be precisel" r.'che-.l with those
recorded for che chlorinated aniline solutions. Althoucil.
there may be some indication of o-chioroaniline formation at:
the ' 5 mg/1 level of applied chlorine (a.= evidenced by absorp-
tion at '--233 nm and ^285 nm) , it is apparent izhat a hichly
variable and complex mixture of products is formed. The nr.-
ture and distribution of thc.se reaction product:-? are influ-
enced both, by chlorine dosage, and by contact time. The pro-
gressive shift of the absorption maximum to hioher wave-lengths
and the appearance of a broad band vi:h in~rna?ir.g chlorine
dosage suggests the formation o" a rr.xtv.re of chlorine-sub-
stituted anilines. In additirn '.7- the formation of chloro-
anilines, it is also possible that scr.c decree of ring oxi-
dation proceeds simultaneously with ring substitution.
The final series of detailed chlorir.aticn '_v:perimcnts v.v.s
devoted to the re-examination of the chlori nc-dinethylaini no
reaction. Dimethylamir.e concentrations of 20 mg/1 ami .100
mg/1 were employed in these tests, with nominal applied
chlo.ri.ie levels rangine ];etv.x--n 20 and 150 ma/1. :: in:-]«-s
were taken at varvir.f time in~.''.'. Is for free rcsidua.1 ch.lo-
rine analysis (by the CTA proc-jdure) and for total f.-hlorine
residual
iodimetric titration) . The results of ther.c in-
vestioatior.s are aiven
12
1 r«
lo .
It is apparent from the data th
rine to dir.othylamine solutions
tion of free chlorine; and (2)
: v. il chlorine ror-idval ,
including 5 combined fraction fo:. ,'.-. : :-; ':-- initial =r.,:>-os rf
the reaction, gradually dim: r.i ;'._:- vi ~ . ::.-'. 7hc- initial
reaction appeared to be cor.plc-';o .--.'-..or :. contr.crt time of lii
minutes or Jess, v.-herea? the los.i of cor; mod residual ehJor-
ine wr s observed to proceed over snvvra! hours. The data
also indicate that the der-ree of c.- lor i nc- u; :.'..(.: i? vari.a! lr,
51 -
-------
TAELE 12
CHLcni::;.TiOM OF DI::KTHYLAKI::E
n.., API. lied
u -!». r " v*or
Concentration ': °^7n~, 7i
, ,-, , , , mol Cl_ ,.
(r.g/1) .-g/ ] 2 (;-
n.rol D.".A
0.
0.
20 21.3 .68 I'
3.
4.
0.
' 0.
20 42.6 1.35 i'
1
f
0.
0.
20 S5.2 2.71 ^'
3.
T .
0.
]..
100 75 .48 2.
3.
5-
0.
-1
IOC 112.5 .715 2.
3 _
5 .'
0.
1.
IDC i:-, .1;5 . 2.
3 .
Chlonr.
cact _ . ,
I-.csicun
J? ' (r^:/1
rr-0*- T7v-
25
5
0
r>
u
0
0
25
5
0
0
0
C
25
5
0
C
0
0
5
0
0
0
r\
5
0
0
o
0
5
0
C
C
1
3
3
8
0
0
1
.3
-
.0
. j
^ C)
0
C
n
0
C
1
,
.
-
0
- 10
2
1
3
5
8
2
58
58
53
/>
4
6
4
2
1
9
L)
3
1
11
C
4
7
3
2
o
y
1
?
i
5
7
o
7
o
5
2
.9
o
.0
.6
* ^
-
. 5
.2
.9
. -'
"i
^ 4
.3
.7
. 1
. 0
^ o
. 7
o
. o
. 6
.0
r.
1
4
4
3
*t
3
o
o
0
8
0
6
-7
3
7
0
0
1
i
C-^
V.
75
3
~>~
f '
25
2 .
.0
.0
. 5
.0
.0
-5
. 5
f u
. 5
. 5
. 5
. 5
.3
0
0
-
.2
0
0
-
_ 2
.55
. 2
Net Free Chlorine
Denand
r.o/1 ' nnol Cl?
nrio 1 n:-'.A
2 0 . b 5 1
21 i
21 vOl "7
_, , n _ ," 2 1 . D 7
21.05
21.1- j
32. G ^
32.6
33.1 -, . , ...
--. , roj.J. I.OJ
j3 . 6 1
32.6
34.1
38.7 '
38.7
4^7 f41-5 1'32
41.7
44.7
75 .48
112.5 .715
150 .95
19.5
- 52 -
-------
eo
a:
o
FIGURE !8
REACTION OF CHLOR!\'E \V!7H DC/E'r.VL A'-/;\' E
-------
anci increases with ii.crcasin.j applied chlorine dosage. This
is evident from Table 12, which shows both the molar consump-
tion or." chlorine per mole of DMA, and the applied chlorine to
L>MA molar ratio. Xote that dimethylaminc readily consumes at
least 1 mole of chlorine per mole of DMA. 7-.t higher chlorine
applications, chlorine demands greater than unity were ob-
served, but a free chlorine residual was maintained. A max-
imum chlorine.- uptake of approximately 1.3 moles Cl~/mole DMA
, £
was noted, at an applied chlorine to D.MA molar ratio of about
A rigorous characterization of the nature of the products of
the; reaction of DMA with chlorine could not be made, due to
the lack of suitable analytical tests. In preliminary exper-
iments, it was found that DMA docs not respond to the chemical
oxygen oenan-J. test. AIL hough it was determined that DMA could
be detected in aqueous solutions by gas chromtography, the
response v. as found to be nor.-cuantitative. Additionally, d.i-
moti.ylani.: ; vas observed to be insensitive to ultraviolet
spcctrophotometric analysis. These difficulties precluded
the positive ident.ificatior. of any products of the DN.7.-chlo-
rine: reaction.
!.;iscussion of Results - Ci'^lor.i nation Studios
One of the r.ajcjr objectives of the experirental program of
Phase I was the determination of the ability of each of the
sol.jcf.ed chemicals to react v:ith chlorine. A corollary, but
nor.etheletjs vital requirement, was tho establishment of test-
L::c procedures by v.'iiich tii.'i s characterisation could be
achieved. It was rocogni-iLx., a pri ori, that virtually all of
thr tost cor-.pov;nds could, ur.dcr tiie apprci^riatc conditions,
.-'.-:\\c', with chlorine. In fact, halogenation reactions are
ai.'ong the nose important in the synthesis of complex orgnnic
cher;icals. ;io'.'ever, many of these reactions do not proceed
to any appreciable extent, unless rigorous chomica.1. or physi-
c:r. 1 Jriviiig foj'ccs are applied (elevated temperature, p.ves-
sure, catalysis, c-cc.) . Obvio\n;ly, such conditions would not
be encountered in convcntion.r.1 effluent chlorir.ation practice.
The selection of experimental parameters was thus constrained
t:o a rather narrow ranee o1" \r\--: .^r.t t-_-r;pv j. ci cures, pH values
:i<_ar neutrality, and d ilutc. aqueous solutions of roactnnts.
1- was anticipated uh.at some of the test compounds, which are
strJal halogcr.ation processes, would
by the levels of chlorine commonly
is oyrurvot.i.or. was confirmed bv the
commonly employee in inch
be essentially unaffcctoc
.ipvlied to effluent?.
ox; e.rimental data, whi.oh indicated that only five of the fcur-
tcv.-n original tost c.i<.:mica 1« wero o':'served; to react with, chlo-
rine to any nu'. stantJal degree.
:.'one of t! e r.lcoi-.ols tested (methanol, isopropanol, t-butanol)
;\ tendency to exercise a chlorine demand. Al-
r.lcohois may undergo halogenata.cn, the reactant
-------
employed is usually a hydrogen halidc (HX). "Groover, the
reaction is co:vu:\only achieved at elevated tcnperatureo, in
the. presence of a catalyst or .strong acid, or in a nonaquoous
gas phase. -:-.s sue!;, i.t is unlikely that a dilute aqueous sol-
ution of chlorine would affect any of the alcohols examined in
this study.
:."o significant chlorine uptake was observed in trie case or
acetone. At first inspection, this result sccnod anomalous,
as it is hno\ T. taut saturated hetcncs a.-.d al.ceh.ydes will
generally undergo haloyenatior. in aqueous solutions. It has
been reported that the rate of halogenation of acetone is
dependent on the acetone concentration, and is significantly
increased by either acidic or alkaline catalysis, as repre-
sented by the following expression :
V = [ (CiU) 0C.O] [G x 10""' -f- 5.6 icTMl-r] + 7[0i!~]]
/here V i? ti'.e reaction rate in. nclcs/litcr/sccond, and the
!?r,:icketcci terns correspond to the molar concentrations of the
respective species. Using this expression, the calculated
rate of c'nlor i r.atioii_of 20 r.ig/1 of acetone at pH 7.4 IF. ap-
proximately 2.2 10 3 mnol/Iiter/hour, or about O.Gv per hour,
In thu present: stuciv, the neasurenent of this lov; rate v:ar?
not coni.'Otiblc v/itii the precision of the chrorr.atograrhic
analysis of acetone nor the OTA rr.ethod for chlorine. Conse-
quently, a chlorine uptake by acetone could not he ohservr-d,
undur the test conditj.cns.
Ben;:cne, and itr- derivatives, toluene, ethyl benzene, and
bcnzoic acic:, also shov.'cd no evidence of reactions vith chlo-
rine. Again, halogenations of benzenes are rather connon-
place in the ch.cniral process industry, but a catalyst is ur.-
ualiy required to achieve ti'is substitution.
It is Ihus unlike] y th.at benzene cr ti'.e alkyl benzenes could
react with chorine in the uilutc acrueous solutions consid-
..rcx; : i: this .-. tuc:y. Bonzoic acid and r.i troi-er.zone vere con-
sidered to be over, lesi; reactive ti'.an benzene, due to the
ol<_ci rcn-attractino and rino-deactivating effects cf the
-CCOii and -X.;_ "r~-u;;s.
In co;itrasL to b<..-nzcne ar.d the derivatives of Ler.zene des-
crii. ed abuvo. ^ he:.oi was deterriined to be quite reactive to
chlorine, ir. dilut... aqueous solution. This result is Ly nr
ricans surpr i:'ir.;:, since t:-c: presence cf taste ar.d c::or-cau.- i
ch.l oro:-hv':-.r.lr .in r .-..- t n j :'i ch.iorinated ..astcv/nter effluents ':-'.
i'ecn dcr.-.eni;-rntod by rsany ir^'estigators 'w' . Th" high rc^ac
ti\\Lty oS" phei^'jl ir aturii utable to the ring-activating clcc
truri-i:e.l.t..a£;inr: :;ro:'<..rtic s of the -f.ii functional crcur.
-------
The nature of the activating group is such that halogen sub-
stitution in aqueous solution is preferentially-favored in
the ortho- and para- positions, with respect to the -OH croup.
This fact has also been confirmed bv several researchers '
(9
Burttschell et.nl.
to.describe
proposed the following reaction scheme
f phenol:
OH
Ring
Oxidation
Note that the reaction proceeds by the stepwise substitution
of the 2, 4, and 6 (ortho- and para-} positions of the aro-
matic ring. Ring oxidation fallows the formation of 2,4,6
trj.ch.lorcphenoi. It is orobable that the reactions proceed
simultaneously, £is well as sequentially, resulting in the for-
mation of a complex mixcure of chlorophenols and the oxidation
products. The nature and distribution of the.'ie products are
doubtlessly affected by such parameters as reaction tine, pro-
portions of reactants, pH, temperature, and other conditions.
Lee has indicated that the maximum rate of chlorir.ation
occurs between pH 7 'and pH 9.
In the present study, the formation of all of the chlcropne-
nols mentioned above was confirmed using gas chromatographic
techniques. The appearance of several unidentified peaks in
the chromatograms also suggested the existence of oxidized
forms. Additional evidence for ring oxidaticr. may be derived
from the fact that phenol exhibited a chlorine demand in ex-
cess of the amount required to completely form trichloropher.ol.
A maximum chlorine uptake of 3.4 moles Cl_/mole phenol was
observed in .this study, compared vit.h a stoichiometric require-
ment of 3 moles/mole for trichlorophcnol formation.
Meta-cresol, a methyl-substituted phenol, exhibited chlorinat-
ing properties similar to phenol. This is consistent with th*2
structural similarities (and therefore, reactivities) of the
two compounds. A moderately activating methyl group in the
meta position should serve to reinforce the pattern of chlorine
- 56 -
-------
substitution, observer, for phenol, i.e., !;y dirc-c~ir.a to t:.c
ortho- and para- (2,4,6) positions. L'r.or. the addition of
chlori.'ie, it would therefore be expected that ?. complex r.ixcure
of chl 'jri r.e-substituted and oxidation r reduces voulc forrr. .
Evidence for thds behavior is manifested by an c:.sorvod chlo-
rine demand of 5.9 moles/raole, which is considerably in ex-
cess of the theoretical reuuirer.er.t to produce a tri--cu!.sti-
tuted m-Crcsol, and by the appenra.."c of at l->"!St eight p"a):s
in the cbror.\atograr.A of a ch lor in 3 tod r.-Cresoi nolutior. . 'Jn-
fortunately , no qualitative icon-;:. ficauion of z'::r products of
ohiori.nav.ion could be atter-.utcd, ;;uo "O the 1 :':'-'. of ccrror-
cially available cliloro-r.'i-Crc'Solf; .
~i
In a study devoted to the oxidaticr. of p-ienol1: ;y c^.lorane,
Eisenhauer suggested the f olln-.:i r:; pro'rai.iv s'T.-;urr.ce cv
reactions resulting from the- ch.lr.rir.cti.oj* of f. f.'.'.bs t: tuted
phenol .
01! OH COO-! CC°i!
ci . - " ci ci.- cccf: 5- C:COH
' i i ; !
1 .... v I > . ^-1
R1 ! R.. -i R __/ Cll-x. . ' C]
" ci ci '
O
(for m-Cresol, K is a methyl group).
T!:'j formation of non-aromatic oxidised proourus, -;u :. ?.F. car-
boxy lie acids, at higher levels of :.;;'ic;: ci.lcrir.e, v:as pos-
tulated on the basis of observed &:.:.. cs ir. u 1 :: r .V.M o "'.-: t absorp-
tion sppctra. This hypothesis is cc -:.~i.- tent vi-;h the rela-
tively high chlorine demands noted for both r.-Crcso.l and nb.e-
nol.
Hydroquinone is a dihydric ph.oncl, containing -c . -: cr^jps ir.
the 1,4 positions. This conf iguvatior. cf tvo :-.i.t:i\lv activa-
ting substi tuenus should even further racr.ify '::.o cicEtnbii:-
z:ition of the ring structure. Inde'e-.; , hydroquir.or.e r.ay i c>
I'eadily converted to y-benzoquinor.o by a varit?ty cf c.xidi/.inc:
.:: ; ifvLously do-vcriboci, the cxif.a ;i j" c :" :.y::r- -p.:; r.^r.o -^ -
bcr.:::c)':;uj. none by chlorine vas cor.'7ir:'':d in this Ktur'y, u^.int'
ultraviolet svoctropi\otonetric pro-'jdurcs . ..-. p ir---.-.r. ly , t: is
is not the only reaction, since a .". '-::'.:' v.r. cr.ior:.".c dcr.arJ of
3.7 .r.olcs Cl?/ncle v:as obccr'-cc. v::. . ch i? arc n tor thar. the-
t;:-. oo -,.-L' ti c.nl requirement for sir':plc r:-:i.:,-~io:-. f- : -': c:in^.;u ir.or.r .
;:.::: C-VIM- , v.-hen a solution ccntair. :r.r: a:, r.p^lie:': chlorino tc;
hydro/u.ir.one nolar ratio of about 1 . _. v:as analysed after ten
rj:u;to? contact tinio, it vas four.c: .-. rcr.t.nir. .~: r rrxir.r- tely
o. ::.-:. aif of concentration of p-i -r.::c.'i.i:. ~:~.' '.'; :. i '."; lei havr.
-------
been expected if complete conversion had occurred. "'r.or.o
observations suggest the formation of other substituted cr
oxidized products resulting from the reaction of c'nlorir.o
with hydroquincnc.
DurJng tb.c preliminary experiments with hydroquir.or.n, the-
formation of a tetrasubstituted quinonc, ohlcranil, .-.a.1-: con-
sidered as a possibility:
0
O
OH.
LI,-'
cj
OH
0
0
Cl
.Cl
I However, attempts to isolate chloranil were unsuccessful, duo
to its extremely lov: solubility in water. It is r.ov belic-vec":
that chloranil is probably not a product of the hydroouir.one-
aqueous chlorine reaction, since the usual method of orr-para-
tion of chloranil involves the use of hydrogen chloride.
The possibility also exists for the formation of a charge-
transfer (-,:) ccriplex. of p-henzoquinonc and unreac^od h"c:ro-
^uinono of the forr.;:
HO
v-- OH
O
The forr.ation of
a: r.earonco of a t
t ; . i :: s c e c i o s
-;:::- colored
is f c n e r a 1 1 y a c c on p a n j o ci
io:^ anc': a bathoc;-.rrr:
-------
in the ultraviolet absorption spectrum . Phononena of this
type were observed at the higher reactar,t (hydroouinor.e and
chlorine) concentrations, which could possibly explain the
non-stoichiometric production of p-benzoquiuonc, as described
above.
In summary, it appears that additions of chlorine to dilute:
aqueous solutions of hydroquinone results in the formation of
p-benzoquinone as a primary product. This compound is be-
lieved to persist either as a separate species, ar; part of the
quinhydrone complex, or combinations of the two. Further sedi-
tions of chlorine, resulting in chlorine uptakes of at least
3.7 moles Cl~/mole are likely to produce further oxidised pro-
ducts, such as carboxylic acids.
Aniline, by virtue of a strongly activating functional group
(-NH2), should also be expected to possess chlorinating pro-
perties similar .to phenol. It is .known that bromine ro<">ces
rapidly with aniline to form 2, 4 ,6tribrornoaniline in high
yield, and it has been reported that, depending on the nature
of the oxidizing agent, aniline may be oxidized to eithci nit-
'7)
roben^.ene, p-benzoquinone, or other products of ring c]oavarrev
It may thus be postulated, by analogy to phenol, that ar.i.linc
nay undergo the following reaction steps when contacted wit;.
aqueous chlorine solutions:
NH,
0
^1
Cl " -Cl
NH2
'
-
Cl. . -
H-
. .. . r en--.
| ... v ' " «
Cl Cl Cxidation
Indirect evidence for this reaction scheme is shown in ''iguro
9, in which the observed chlorine consumption by aniline, or.
a molar basis, is noted to be quite similar to -phenol.
It is also evident from the experimental data that t -.
rination of aniline results in the.generation of hi
plex reaction products. Ultraviolet spectra taken a-, i:-:--
ly chlorinated (0.5 rng Cl2/mg aniline) .solution", (see ":. rjrc
16) suggest the production of a mixture of chlcroar.il ir.
-------
Higher applied chlorine- levels (1-2 rag Cl-Xme:; seem to pro-
mote the formation of oxidized species. t.'oto the '_".' absorp-
tion spectra obtained on a 10 mo/1 aniline solution, co v.-hich
]0 rog/1 of chlorine had been added. The disappearance of
peaks in. t^e region of 230 nir, and 280 n~ indicate a loss of
aromatic structure, normally associated with those absorp-
tions. Moreover, the appearance of a single broad bar.d sug-
gests the formation o£ either a partially oxidized r.or.--?rc-
ruatic ring or a conjugated straight chain molecule that re-
sulted from ring scission.
There is also evidence for a continuing sequence of reactions
that proceeds even after a complete loss of free chlorine-
residual. For the 1:1 solution described above, no free
chlorine residual was detected after a one-half-hour contact
time, yet the UV absorption pattern continued to change over
a two-hour period. This may be indicative of the production
of intermediate species, which may subsequently undergo struc-
tural rearrangement. The resulting broad band at 240 r.m was
found to persist during UV scans taken over eighteen hours.
The position and the shape of this absorption is quite sirr.ilar
to that observed for p-benzoouinone, a possible product of an-
' iline oxidation.
i
Dime thy la nine, a secondary aliphatic anir.e, is also known 'TO
participate in halogen reactions, leading to the formation of
r.iono-Nhaloamines.
In a previous study devoted to the rate of formation of chlor-
(12)
amines, V.'c-ii and Morris "" indicated that only a moncchloro
derivative results from the rcac.ion of chlorine and dimethyl-
amine, corresponding to the. fclicwinq stoichiometric represen-
tation :
H Cl
CH3-u-cn3 4 ci2 -» CH3-:-:-CH3 + uci
It is evident. Erom this reaction that the stoichior.otric
chlorine requirement ifj one mole per mole of D.XA. 7;-.. results
from the present study generally confirm this stoichicr.etry,
hov/cver, in cases of nicher a::
clomands greater tlian one. mole
.ilied chlorine dose?, I':.lori.no
)er nole wore observe;";.
Marks has stated that the monochloro derivative of di-
methylanine (Nchlorc D.'-'.A) should be amenable to analysis by
the acid iodometric titration procedure, which was utiiiL-cd
in the supplementary L\"J-. chlorin.ation exrerir.en ts. Indeed,
an iodometric chlorine residual in excess of thr- frc-c chlo-
rine residual (CTA) was observed in the early fta"C:=. cf the
reaction, suggesting the initial formation of .-/.-~c cr.loro-D"/-.
compound, probably '.'.chloro i)'.'.:'.. Vhe subsc-cp:cn _ lc^.~ of com-
bined chlorine residual indicates chat uho compound is either
unstable, or is lost frcr, the systom by volatility tier..
-------
In summary, it has been shown that five of the fr.
ginal tost chemicals wore observed to participate
tions v/ith free chlorine under the moderate co-.di
ciated v;ith conventional effluent chlorination pr
These compounds are: phenol, m-cresol, hydroauirio
and dimethylamir.e. In some cases, it has been po
relate the experimentally determined chlorine-roa
these species to their respective molecular s t r u r:
exarr.ple, of the nine aromatic compounds examined
only those possessing "ring-activating" substituc
were; noted to react v/ith chlorine in dilute ar-uc-o
These considerations should facilitate the pro la r
acterization of the potential of other similar sp-'-
undergo chlorination reactions.
The information developed in this phase of the st'.'.
indicates that the application of chlorine; to dilut
of any of the chlorine-reactive chemicals results i
complex mixture of products. It is apparent that t
and distribution of these products is dependent or.
of phrameters, v.'hich include reactant concentration
temperature. A list of the prohable products of ch
has been, assembled and is presented in Table 13. T
tiqn was compiled on the basis of the experimental
rated in this r.tudy, supplemented by various litera
sources.
.eon ori-
asso-
ur
i
ti
tice
on
bio
ct-vi t
I- ;:r <.-.-:.
in '--.'r.r:
r.t ':rc
u:-. ro'
i.n ":;.-
uti o.-.r;.
cle
e sol
na
s, pH
lor I:-
). i :: t
t; ita
ture
arly
ution
i.f:h]y
ture
iety
, and
.?. tier.
-
One of the objectives of the experimental program of Phase I
was the selection of five products of chlorinaticr. for sub-
sequent studies. The chemicals initially chosen for furt!.c-r
examination were 2,4, G trichlorophe-.iol , 2,4, G trichloroar.i I i r.c- ,
4 chloro 3-nethylphcnol , N'-chloro OM/. , and chlorar.il. Tho
original selection v;as made on the basis of availa;;]?.- infor-
i^ation relating to probability of formation, chemical rtn:il-
ity, and commercial availability, or ease of synth^si :;.
hod
re-p-
' r, ti -
Specifically, 2 , -1, 6trichlorophcnol was cho.c',en since- i
been identified as a product of phenol chlorination, a:
resented the final reaction step prior to rjnc oxidat
Its ex is toner had also been demonstrated by pi-L-vioj;:
gators, who indicated that it was difficult to bi.c.lo'ucol ly
f 1 T 3 '
degrade and was relatively toxic to aquatic oroanisms -'-'"'
Trichloroanilino v;as selected for further study -.eca-.r-e of
tho chemical sir.-.ilarity of aniline to phenol, as v/oJ 1 a.% a
strong theoretical basis for its existence. Virtual j / nr.
information i:: T/ai].?.:;lc rr-:.. ;ir.; tl:«_- ::c'r.avior cf L: i - CT-
oound in Liolc;-: cal svEtcmr.
The only commercially available chlorinated product
Crcsol v;as 4-c:J.oro3-jiietJiyl phenol, and a.~ little-
r.LoMt the products of m-cresol c:~:lorinat-j.f"., Lr.is c
v.as carried ir.tc the Phase II studios.
- 61 -
-------
v:.:-.!.}: 13
oiiABL!. rnoDuc'i'P or c'.;i.oi:r:.ATio:
o- oh lore-. "o no J
p-c'nlorop honol
2 , 4-dichlorophcnol
2 , o .1 i.chlcrophv.T.ol
2 ,4 , G-trich^orophcnol
non-aro:.:a t.i c oxi.dat i on products
.-Croso'i
2-chlorb-3-r~l:T7y.Lpr.cnol
-',-c;;iori.. 3-:".orhy ] plior.ol
b -c: i lore- 3 -;;-.o t hy 1 phono 1
2 , 4-cl i.c''.ioro-3-'. ... thylpl:-:>;~.ol
2 ,0-cic:',] oro-j-r.othylphcnol
i f 6 -c! i ci . 1 oro- 2 -r.o 1 1 ;y 1 pho no 1
2 , , C-tr j chJ Dro-3-r.otliylphcnol
r:C;-i-;iro:v,aui c: o:-: iuation produc ts
l'yuro;;ui nono
.1 inc
; ->.::. lore 'in i .1 in.c
2 , ' -c1. ; ci: lo re- .in i. 1 i nc
2 , i-.-vliciilovo.ini.lino
2 , T , l -Lr.i cr. lovoaiiiiir.o
r.or.-f.rr': lat.i c o::i.:.ati or, products
Of Lr:o p.'-ii.-.c-.ry te^i ci.-:-:^a].; ^-::-..- idorod, di:".i:r\-ive ri.ol.c~ic."1.1
tro.at:..er,t intact. Sir.Cvj ocv.".o cv: d-,":'.cc ^::i?tcd for tr.c for:
Vioii of i.i.e r'onociilorc di rlvativ. , '.'c:.lrror"..:-. v:?.; d^crcd
:iiii t;iL j.;.: for fu.r':-.^-r :-.:..... .' 1 _:..".":. Xc:. J or r1"! -;. v;as ".r-v
.ivaiiaMe ccr.r.;c;rr.i .ij ly, i v vc --' iv.ain^Ily antiriratcd ':iat
it;; synti.onis ccv.lt.: Lv a.-..:;\\.: ; /. L::O la. ovatrry. iic-> vc
v.-i:on f;ulu;oi.:uo::t stUv;: v/r. j "id: ca tod. ti.at Xc:. 3 cro-r:7 .--=.-
oiionri c-Ti. ly ur.stn1.. J o, {..::* r;;cc:!o:- v/as drrrpcd frT. fv.:"::-.or
connidi.-Vv:ti-n.
-------
Jhicranil, although not srovn on the list of probable pro-
civ.cts of chlonr.ation, V.XIG initially suspected to L^ a pro-
'iucu -.)f hyclronuinono chlorination. As such, it v.-as crinin-
aliy chosen for examination in Phase II. When additional
_:::: erir'.ent;s failed to establish the existence of cMoran: 1
as a reactior. p/rcduct, p-benxcquinone, a confirmed product
of the iiydrocui none-chlorine reaction was substituted in
ill.c: follovi:-io studies.
63 -
-------
i
SECTION' vrri
. P.ESPIP.OMETER STUDIES - L-HASE II
As a result of the Phase I studies, 5 corr.iour.cls v:erc selected
for further study in Phase II. The selected compounds were:
2,4,6trichiorophonol
4 c r. ioro3iae thy 1 pi ieno 1
chloranil
2 , 4 , 6-t.r j chloroaniline
. Kch] orod.i.r.c thylainine
The first portion of Phase TI was the conduction of resniro-
meter studies to ex?.nine the possible inhibition or toxicity
of the selectee; confounds to an operative biological system.
Experimenta 1 Procedure
The respiror.etor studios wore conducted in a modified Karburo
apparatus, known as a "Cilscn" respiromctor. The system dif-
fered from £i "V.'arburn " ' or. ly in the method o'C pressure diffor-
ence measurement. The cor. Lrol systci". was an inocculatod syn-
thetic sewage of th.e composition described in a previous sec-
tion of the report. The tost systems contained the same in-
. occulum and sowacic, plus the desired concentration of the
particular substrate. In most cases, a substrate concentra-
tion range of 1- iag/1 to 100 r,:c./\ v.\is stuc j oci. Tor cor.parative
purposes, one- exporir.cnt v.'Cis conduct.r:"a t ion on -Ich J fro3
r.eth.yl pii'incJ . "'>:>. i- .'< strace it: r.: Icily j r.hj birory at 2ov:er
concentration s'iC'r.c; 1), stronr;j.y inhibitory at 50 re:.'], and
apparently v.ox.ic at 200 r.c/1. ri.cur-o 23 ;-.-rescnts data for
c'r.loranil', .:;-.: cl-
I ions v: ere not s t : i ,1 i c
tnc .Material as r.cn .1
h.ibitrry at 10 mr./] . Hinher concentra-
hecauci.. of .''.oJubia i ty liir.i tations .
line dt ta .-ihov/r. i:: !-'i;-urc 2-i svu-crsts
.--.i.:;; up Lo 10 :-.c;/l. Solubility
ed investicatioi. aL hjci-.or con.contra-
^;ch 1 orcicii' r th.1.' L ar.ir.o '.-:;<-: or i.'.; ".a] *./ 'jcv-sidiTriHt fcr P~x.fi" i'ut
an invcs ti'.:ai.ici": shcv:cd thai. Lhc ror.i-ouv.d wa?; i, p. n t-a i !:, and
Preceding page blank
-------
. J
o
200 r-
130
I GO
i.J 1-10
!20
il SCO
x
o
eo
20
,?
LfGFND
i"- !CP .0 m-; /I f
';'. 00': :r-.';:. (^c.'J
'S "1C? ! :,')/! 4 S>
1 KP 10 mg/1
SEW.
0
IRI3 250 375 500
625 750
Yi.VZ-HOURS
875 OCO 1125 1250
FIGURE 19
-------
200
o
0 125
. ._J_l_-
25.0
375 500
6?. 5
T;,-.-;E- HOURS
FIGURE 20
EFFECT OF 2,4,6-iRICHLOROPHENOL Of-i A MIXED WICROBIAI. POMjLA
-------
7 CO
i MO
-. 160
o
a 17.0
o
t:J
'r GO
;. C:.1.'1 in-.;/; i Ui.'J :.["-v
j. C.Mr' luvj/i t G'iN 'f'-v
-: CM^ IOM..J/I > StN SEw.
GKED 5%
125 250
57.5 500
62 5 75.0
TIME-HOURS
875
!000 1125 1250
FIGURE 21
EFFECT OF 4-CHLORO-3-METHYLFHENOL OJ A MIXED MICRCBlAL FOPULATCN
-------
.J
V,
O
n_
X
o
llj
>
I -
-t
. 1
."3
o
100
160
140
120
100
80
60
'v
20
0
125
250
370
500
62 5 750
y;v£- HOURS
87 h
;COO
1250
1
FIGURE 22
EFFECT OF 4-CKLO!-'0-3-f/.ETHYLPME:-.'OL OM A ^iXF.
-------
LF._GFJ-;D_
7i;, CG-'/r-ici'lSr;.' SF.w)
/>. CHLOR-V..IL l.nrj./!fS-'.\' i
CMi.CRP..'!_ 10 -T.-'! 'S
125 250 375
500
62 5 75.0
TIME-HOURS
875 1000 112.5
1250
FIGURE 23
EFFECT Or CHLOKANiL CM A :1.!XEO f-»!CR02!AL POPULATION
-------
200
180
-' !GO
c
LU 140
<-t
k 120
la 100
X
00
[i 60
ci 4°
20
0
c
-
- -~ *"
^ -"' "" ^1^^=^=-
^^^^
^^
'/
^^< '. , . ; i
> 125 250 ' 375 500 &25 750 875 ' '0
TI.YE- HOURS
'I COT
,.; 1C-'-
L_
'000 ii.25
FIGURE ~4
u-fLcr cr 2,4,6-TRio:!..o:;oAMiLir.:[; G?J A VIXFD \;ICPO;'::.M. s Oi-u.-'Mirxv
j
-------
degraded at a rate sufficiently high to nake it? persistence
in an effluent ir.nrobab.le.
-------
:-;. -.TIC BIOASSAYS
PHASE n
r.v- Li. .-.. .;_::. e l-iocr-says were conducted to determine the four-
:'. -.y :..-jc: : .... - -ol --r :: ncc limits of an appropriate test oraanism to
:_.-.c: ':'::_: r:-. pounds . Po? purposes of this study, the TL v:as
o;:t:::.lis:v.-. tc within one. order of ".annitude . n
siaLj.. i.ic-:t:;sciy^ :.\ore conducted according to.
.:. us '"' :.rojedurcs .
Ch lor ino-fr _£ tap water which had been vigorously aerating for
f.:o d .-.;/:: ;..-::.or to Lh<- start of testing, v:as used as dilution
..j.tor. I'':-.:, .j «..": ;:'.-. ; :ov.'s ( p ; n-. ,^- - . h o I R s D i o -. o 1 a s ) '.core- -ist-ci as test.
.'. :.:.\:r.;. :;:../.. ~.".. r c v~ '.'. Cil Ii.i. ni-i.^s aquaria, nacft holciijic; ten liters
o: v.'aLur, v.-::e u.soc.: as bionssay containers. Three separate
soric-s cf .;-atic !)ioa«s>-.ys v/ere conducted.
'no
is were tested at three different concentrations
:: J ,-i, Ct.-Lchiorophenol (TCP) and 4chlcro3-i-ethyl phenol,
}, 'nth .it TCC, 10, and 1 r.;o/l; 2 , -'.,6trichloroaniiinc
.'/ , ..-; II1, 1, and 0.1 r.K.r/1; ar.d chlcranii at 1, 0.1, 0.01
::.-.:/1.
The two ;:.:. no 1 i.c co:apou:.cs were added to the test containers as
aqueous s....: ..-j.c.ns . 'I'-.-.o containers, each holding five test fish,
recc.: ;.voc: .:.-^ i:i;.nre:v ra1..inn, :.ri:iq:.nu the total to six containers
ana thirty fj:-;t fash, for each of the two compounds. In addi-
r;.or., two cr::-.._ai!:v..rs, each holding five fish in straight dilu-
ent, water, j-jrvud a>: controls.
"'ho other -:: f.-oinpounds , 2 , 4 , ctr ichioroaniline and chloranil,
:,c:cauf;c ;.:" t..-_ir rci.-itive insolubility in water, were added to
t/.o tv.~t ..-,:.-_ =; :ujr:~ .is r'.oth.anol solutions. The facilities for
\.-jH r. -:r.-.;; -.r^j.. - ' t:i'.: two cr.;,.rounds -.'.ere exact?.y as described
f^:~ t:.e --:'. : . r.-'i-ic corrpouna?. Additional controls had to be
.3^': ".] t . -_ .: ;r-.i:-.'.% the.- ef foots of the r.ethanol that was added
w:.th :':.: r:r ~:::\.\r-. Si:;-,-.:: the chioranil ir. -cthanol solution
-..--::- c.:: .'..?:. .-:.;::.-: :ts to rnsui': in a 400 no/] -athanol con-
-:.:.. r-.t: . - : :. -/.c- : :;: .-ortai nor s v:hon the chloranil concentra-
t_'.;. .:.!.-. ;. . : ., ,.-.- ;:c:v..roJ ccnt.ainers, eac:i holding five fish
in ::'^J .-. " .v ...;:K/i. '..n ::; i.ue.-'.t, w_-re prc-.\M.ded . T:.e 2 , ! , 6 tri-
c-.l-r^-.":;il : --. ;n r:-.j:::.a:;.^i nclxition resulted in an 30 ir.c/i ir.cth-
;;ncl co::-:-'.-. _;'.;~. i.o:; who:i Ll'.e TCA. cor.cr.ntration was 10 rr.g/1.
T..-i:3'.-r.-..'..;, . -Tiid-1 i")n::l control container." were set up for 80
tosti p.«, th.e -.vercent r.ortulitv of
-------
The 96-hour median tolerance lirits of fat.:.one! nir.r.nvr. to eac;
of the compounds tested .'./ere : 2 , 4 , fctrichloror-hor.ol -1.0 -.1
me/I; 4-chlore-3-rr,f thyl phenol at <.l >.01 r;c-/i; 2,4,G-tri-
chloroaniline at <10 'l.C me/1; a::d chlora.-iil at -'1.0 - . 1 n<"/j
Tr.e tast results arc presented in Figures 25 throiu:': 23. The-
chloranil test fish exhibited a somewhat ur.urual reaction pat-
torn because of the insoluble nature of the cr-npounc'. Despite
precautions, the chloranil flahed out when the r.cthr.rr:! solu-
tion nixed with the-diluent water, .and floated or san!-- to tr<-
container bettor1.. Sor.e of the test fish were seen ir.oestir.t-
.these solid particles, rredor.inantly at the 1 n^/l concentration
level, and exclusively at the start of testinr. The r'-r ai nine
fish did not do the sane, and consequently, survived the 4-day
period. /.t the lover concent rat ions, onl'- a f-ijv r3'r;h wore af-
fected. !!ore fish died at tie 0.01 r.o/1 concentration than at
the 0.1 ng/1 concentration, probably because r.oro solid-parti-
cles of chloranil wore ingested by those fish.
Series Kunbcr Two
One conpound, p-hcnzocuihone, vas tested at throe concentra-
tions, 10, 0.5, and 0.1 me/I. The conpound vas dj.s-ier.red to
the test containers as an aqueous solution. hxacniy the For.c:
facilities, including controls, were used for be ri ;:<"'.'; u in one ar.
'.:ere used for each of the phenolic conoour.dn in Sorioc f.'un'.-'.-r
Ore.
All of the test fish died vithi-. ti'ie first day of tentinn ovc-r
ti;r entire concentration range, while 'ho cort-.rol fish survived.
liov.-ever, the control fish r.ortality vac too c-.rcntz nftv.-r 4 d-ys
to sacisfy Standard '-'.etiioc's criteria- for i-trst vnj.ic.ijty. It war.
thcucjiit that perhaps ti'.e benzoquinonc var-orizr-d fron sone of
tr.c test containers and contanir.ated the control diluent waters.
Consequently, fa th.ird bioassay vas sot up to ror.tucly bcnzoqui-
nor.e. In this study, tiic control containers wore well separated
iron the test containers. Despite the unsatisfactory '.chavi.or
of the control fish, this test did. yield valid i:n'"orr?.tion. The
reaction of ben."oquinone was inr-cdiatc ar.d vic-lc.nt. It war. dr-
cided, tiierefore, to tost t::e cor.pound at'lo-. or concentration
levels in iiioaysay ^lurbcr Three:, startir.c with 0.1 nr/l.
S e r i e s '.' u r V:. or 'i h. r c: e
Iier.2ocuinp:io v/as tested at three cor.cc-ntrations, 0.1, O.f'l,
0.001 r.a/.l. The sr.r.e proccdurc-5 as ur.od :n riicarf.r" '.'.\\r.\ ."
v/cre follcvec:, except that four fish per C'X-v.:a i r. r. r :; rr- ii!:>:-r
The results of Bioassay l.'unhcr 1: rce a'-c rror.c.ntcd Jr. Pirrurc
?<-
-er.xr.-runcT.t: v.T;rr :-'
vithin the first tvc day? of testing, v..i]o Vho J.'-.vor concor.-
tracion le\'els had no sicr'.i f i~ar!t o-"frct. \:.:'-.-~\-''.-, :.':<. c<"-r.'i'ol
-------
'////,
10? r-
I I
I rir»
! !
I I
-:GU~E 25
-------
100 r y
! >
7 c: A
10
.VG-'L
,,LJ
TIME- DA IT-
'TO
80
(.'0
.' D
c*
y.
'- 50
T
75
FIGJHE 2S
RESULTS OF STATIC BiOASSAY SERIES NC.
-------
: 00
70
. ' ''/.
70
70
j
I
~i "*
~,\.'//,.\ '/. 1
' C!:'. ORA\ '_ ':
'00
C
C
50
i
RESULTS 0^
F;GJRE 2?
"A';C L:'OASSAY SZRiES NO.
-------
f
'/i
1 I
.". \' r
-------
'00
25
r^-v^O'
! !
P 7 5
i i
n -Pr :roc'ji\io\r o.cv VG.-'L
i i
I !
RESULTS OF 3;
FIGURE Z9
ASSAY \'0. 3-
-------
fish r.e.-'.in exhibitor] hiqj- mortality after - -Lvys and this tost
did not r.'.eet tiie Ptr.ncU.vd Methods criteria for validity ei-
ther. Hovever, cor.troJ "Tosses \.ert- net a? drastic ?.s in ti.e
provious ;>ioar,say, r>.r.d the lover crnccr.tratiop. luvcl? c-f the
compound j;oer;od r.cn-toxic. The 90-hour /L.,, :T;-; a;.;ly has a
value of cibout C.OOL> to 0.03 nc;/l, c;s soon ;.n r'i rv.ro. 2?.
Suinnary
Five ci:l orinatod organic cc-v.youads .'ere vc^-tec frr tcxici.vy tc.
Dc'ith.oad iv.ir:;iovs. Or tl'.c five', four 90-'.-.cv.r rtc-.ian tolerance
l.ir.ics v:cre cetorn.ir.ed. rcr the fifti: ccnptjur.r., n-Lcn^oru ir.o.".e,
a probcible toxic concentration \-AS r.etc-rnir.cd. Vhcsc values
aro presented in TaLle 14.
T;,DLI; 1.4
iiHSl hTS OP STATIC BI
Compound Tested OQ. hour l"rr: Pan^c-- (r.a/J )
2,4, 6-tr i chloro::-honol 1.0 - 0 . 1
2, 4 , C-trichlorcar.ilint. 10 - 1.0
1-chloro 3--r.ethyl phenol . C.I - .03
chloranil 1. - 01.
_ ... - Pro:;?.:: ie 'i'ox'c Concentration Level
Cor.pou.nc * c s ccci ' ..... /;
p-bcnr.ociui.nc'r.e
- SO -
-------
hcvi'o:: :.
'.. '. . .:v. i i" P. o: ; Usdy C <>" ound ;
. .! (::.'.:'.:<. .;'.:. --i.c.v < > !' sl.VMv ] ini Lat. j or.s , it v:ns ror;uir<.-c uo
;:.ii, '.h> i'h.'is'.- Ill studies to three ccrv>ov.r:cis . -4 chlorr, 3
..:_:.] I'iu-nol '.'.'.; o l.i r.i n.u U:d in vi ".; of t::o e::pcrirr.cr.tal rj.'il:."
:..:! v1.! i :..- t:.. ;; w.,.it.t.: r ; >"o! rii-iJif/ of occurrence! of tiiC
': , ' , L -'. i' i .': ! i.m- i.er.ol . Ci;!ora:ii] vas o.l ir.ip.a V.P,.; in vi^v <". f
.- v--\i ri^-.uvL, viLiustioi; au tc ^i'.othcr the: cor::'cu:'.fi car actually
'e foiT.'et"; uricior troa'crx r.t plant cc-r.cii tions . 7 he su: str:>tos .
> ": ooted for Piuisc III study verc:
11 , ! , '- 1 rich, Lorcnr. .i lir.o
p-! en?.oc:uinonc
.
: .; j.;- ,;.i.v ,::^;ay i.!'.\ i.'.'".l_ i.oto-
: cf..!1^: j. tior.t: . Ki'jcilly , a uroaricr st.udy conr;idori .p. 9 r.npy
.ct.\ ;' i.. e food ch;;:r. over a lor.cic-r v-crioa of tire voulci
.;;,!. ;. . '.:..: r. s! ^:;d ' ;\fi ] : '.. i .-:; :-ev".i j red for c'-nci'.ic-t-
...:'< ,:. : ; !' ..::'. '-.jy:' ':;.: s;.,ir:]v OU r :: .'! i 1 (' d i.r.\'C';~Lif":i-
'..; HL . . i .-; r..it.'.i: i- . 'i'h i f, :V.T ;-. i_-ct C::OPC to consider an in-
;'ec. i :il o r.caic1 .ij.o;.:~nay , fiov. th I'ouc-;', ecosystcri that crulc;
v-i'iv. Li d .it a rc.'.:"o--.ai Ic cor-t, i..ut v.-oi;li; provi.io hetto:r
;.'. : .:' . i-.-p. (:. or'.'! !/'i!,i \>ni.,.: J riv~.;io:'.?(- at sovcral trooj-io
.., v.i. ..;! ,ir : ' :!:t!.;, nacroi r.vo rtcd. ra ties , and r.icroscor ic
..;.-... .'.-..'. in LI .;'.-. V^:~L. i .:-.\T,V :;:it.-,:>:;. 'iVcr.ty gallon aouar:a
;;' f.i.-.- >. -.;.:, va!,-; .r/r.tcrr, voro c: osen aF ti'.c ^tv.cly
_ : ... ::-d /. Ui.«.ly ; ." riod:; of .':; ; re:-:: r.av el y one r:cnth por
:.; .::-.I.U,CL . n v.ire cor.r;.1. r:«. rcci.
-------
SECT I OX XI
"RriPLLTS OP PHASE III
Fish
Fathead minnows obtained fror, a local ha tciicry wore omplovcv.
as a test organism. Die-off of the test fish occurred in
most of the experimental unit? during ti;e background ace li na-
tion periods and in the control u-.-.i-.s durino t! e bioa^savs ,
complicating data interpretation .
Figure 3u is a graphic representation of the nunber of tost
fish surviving versus time, for each experimental unit. In
Table 15, tiie duration of the study is broken dovn into con-
vcnient periods ana the percent mortality is gj von for eaci
period and for the whole study.
i: si:.".- :\KY or FATHTAU MINDOV; ..-'
Control '.'P.: ts
1 in i \- -J:-
\.i I I a. l« ,.
1
S
ilnit- {
n thin time until t!
beginning c-f: ''ay, a hic'i rate of die'-cff occurred, reaci ir.;-- '.
i only be attrii. utcd to d.iseare, or -sc'V.
TO contamination by ary of the c.->r"-eu:-i':-
in one case. Ti~.is ca
natural cause, since
V.-.TH sliovn in soeetrc-
iotop.et) i c ar.al'.T^es .
Preceding pass blank
-------
30' v
10 -
. !
;.:c
I 0 .--
i
r^ 2 o -
10
uvr
SEf.'^OOUIN'ONF
','A ' 26 IS
O 30
20
i o ;-
i ' I i
15 10
30--T--^
r\ f\ _
cvj :
10
U\'lT ',
CCf.'TROLS
261 15 10
OCT: NOV ' DI.C
1 1
PCS . MAR
Af'R . './,<
K.iGURE 30
FATHEAD MINNOW 00='JLAT!0\S VS. TIV.E
-------
The total percent mortality of the control fish through, the en-
tire duration of the experiment varied fron 22^ to 37". There-
fore, no significant toxic efl'cct r.ay be attached to lov:or .-irr-
ealities in any of the tost units.
TCA shov.'ed no effects on the test fish at any concer.tration
level. This is consistent v.'ith the findings of the four-day
static bioassay, since the 36-hour TL.^ as determined by that
test v;as not exceeded or oven reached in the flov-throuoh
studies.
Bo-jnzoquinone and IC'P exhibited no significant effect at either
of the tv:o lov.rcr concentration levels. Ever, at the second
concentrations, w'-ich compared v.-ith U;e DC-hour Tl._.'r:, the
mortality was less uhv.^e test units than in the controls.
This' v.-ould seem to ' icatc either a greater resistance of the
test fish to both of these- compounds v:hen in a more natural
environment or the degradation or assimilation of the com- '
pourv.i5 within the aquaria. Spootrophotomotric measurements
indicated that some reduction in concentration did take place
in th;i up.it:;. A comparison between aoplied concentrations and
observed concentrations is presented in Table 16.
TABLr. 1C
COKCENTPJ.TICK'S (mo/1) OF SUBSTRATES DURK.'O
VAKIOUS Ti:ST PERIODS
!'cr i od
Applied .'ieasured Applied ."eas'.ured Applied :-'ear;ur.. ;-.
p-ben/o- c<0, f) 3.103 O.OOG l.lr; ^ " -
quinone
2,4,6-
tric;:lor- , , ,, , . n , ,.-
... - - O.H O..I4 O.iJD
c^an.i ii ne
(TCA;
2,4,6-
triciilor- ,, .. ,,, . , -
'(TCP)
( 1 )
helov: mcr.surcable range.
r\ "> 5 ~ 1 -, 1 " 5
'/ » .- J *T JL ' J.- * " -^
V.'her. the third concontratien levels 01 bi.M'.^o.juir.ont1 ar.;l TCP,
values ten tines create;, than their 9f>-hour Tb_'s, v:ere dis-
pensed, the reaction of tho ^est fish v:.-s ir.r:.edi.-.tc and vio-
lent. Kithin a fev: day;- a ft':r r-tari-i-.cr r.!-ese bioasp.->.ys, 100?
kill v.-as effected :n all four '-e-;t units. Compound moasure-
msnts indicated that u'..ese icthal ccr.cc: tra'. ior,s v:ere sont?-
v:hat less than half of the roninal levels delivered to the
aouaria.
-------
Vascular Plants
The growth' of. two vascular plants, L u d v I g i a and Andc ,'.
was studied in detail. The total length of all Luc'v i
stalks in each test aquarium was 35 to 4G inches at the be-
ginning of the study, and had increased to 75 to .100 inches
by the middle of May for the control, bcnzoquir.one , and TCP
units. The plants in the TC'A units had grown to over ICO
inches L>v tlvs end 'of Kay. None of the to.st compounds affected
ti;o hr-alv.n and :jrov:th of L u a v i r, i a at any concentration level.
The: growth rates of. Anacha r i s varied so extremely anonq the
aquaria during the background period of the study that compar-
ability of the units to one another was severely hampered. 7,s
stated in Table 17, the average growth rates varied fron loss
than one inch per week to no re than seven inches per week.
Prior to the start of the bioassays, the Ana char i s plants wore
re-distributed among the eight units so that each contained
approximately the -sane number of inches of. total stalk length.
TAI'.LC 17
VASCULAR PL/.NTS GP.OV.'TI!
'unit
Average growth rate in
inches per week*
over background period
L u d v i r: i a A n a c h a r i s
Control unit 1
Unit £.
Benzoquinone
Unit 4
Unit 5
i. 1
0.9
) .0
0.9
0.3
1.7
Average growth rate in
inches per week*
over bioassays pcriodt
L L; -j v i n i a Anacharis
3.5
3.8
2.4
5.7
1.8
3.0
'Jn.i.t 6
Unit 7
0.8
5.G
3. 4
D.4
6.3
Unit
2
3.
1. (>
1.3
5.7
4.7
6.9
5.1
*vhat is, the total number of inches of new grow;
biited by all six ruin stalks.
-l-'roni beginning of: Con:. 1 no end of last Cone.
contri-
p a c h a r i s in
:;;!:jng the bioc.ssay?, the aveiraae grov.-tli rates of
ci'.o bonzoouinonc test aquaria were the slowest:, s?:ovine a de-
crease from th.e background rates. Ti;e growtii rates increased.
in all otncr units. Hcv.ever, tl.e control plants in Unit rl
>G -
-------
still exhibited very slow growth, less in fact than benzoqui-
p.one unit -:5. Also, ben^oruinone 1."p.it ^4 suffered from the re-
distribution 01 plants, sir.ce it was given some of the slowest
growing stalks from Unit -=1.
Ir, summary, the inconsistancy of Ar.acha r i s growth proved to be
too great to use this plant as an indicator of compound effects.
It was observed, however, that nor.e of the compounds affected
i-.he general appearance of outward health of Anac ha r i .;.
Benthie Macroinvertebrates
The nacroinvertcbratc populations underwent so sharp a decrease
over the course of the background Declination period as to ren-
der them unsuitable as test organisms. .As shown in Figure 31,
out of 93 individuals (excluding 7u L-. i f cx) present in each, aqua-
rium at the time of initial seeding, an average of only Ib per
aquarium could be observed :'n rr\id-.".arch, a decrease of 84 r>.
The average Hydropsychi dae population decreased 90C,, fron 31
to 3 individuals; A s e I \^^ decreased 771, from 13 to 3; Ec topr i a
69%, from 13 to 4; Gy ran I us 79%, from 14 to 3; H e I o b d c 1TTV 86%,
from 17 to 2; and F e r r i s 5 i a 1001.
Several reasons for this drastic decline can be identified.
Some of the orqanisr.is avoid observation, either by burrovino
into the foenthic sediments when observations were made, or by
burrowing into the sand matrix. So~c of the oroanisms arc
consumed by the test fish.. The fish were not the only preda-
tory stress on the niacroinvcrtebr-?.tes. Asellus is a potential
;.>redator op. all of the other animals, arc: lie I obJc 1 ! n preys
upon G y r a u I u s and F e r rIs s i a . Since no effort was made to sep-
arate the various typos of r.acrr-ir.verbrat.es from one another,
He1o h d c 11 a and A s e 1 1u s may have consumed seme of the other
orcjanismr;.
Adequate food supplies may hr.vc been lacking. Specifically,
the microflora and faura populations may have been too low to
support the macroinvertcbrates during the background acclima-
tion period. The lack, of nutrient- in the diluent water may-
have too severly limited the rapid crowth of algae necessary
for a productive ccosysten.
Fish, waste products and other de.tritus built up in the acuaria
with cime, scttlir.r: on the ber.t.r.os i:; thickeninn sludge layers.
Perhaps the cases of. decomposition released through bacterial
decomposition of ti.is waste proved injurious to the macroinvcr-
tebrates. Tr.is VuTctcrial activity also rviy have doyjleted the
dissolved oxygen resource at the benthos-water interface.
I-'inally, tive marked drop in the ca:;dis f]y larv;; popu 1 ;u:ion v:ar
influenced by .1 special factor unique to Hy d rop:- y c h i doe , .'. t. s
annucil metamorphosis fror larva to pu"-a to adult. Only t!':O
larva nay be used as a test crcanisrr, since it is active* in
-------
1--
L i
-------
-water, while the pupa is in.nob.ilc within its cocoon and the
adult i-s a terrestrial organispi. In several instance-?; from
February to March,. innobile cocoons and adults s'truoglir.o to
leave the water were-: observed. A significant portion or the
'I'.'/dropsyc'-iitisc popu 1 a t i on may have undergone r.etariorpl'osis
at this t i. ne .
.''.ic'ros.ccric Flora and Fauna
A total cf six. sets of population, counts were race, throe- of
two-wee/: populations, and thr ;e cf f our-weck ~c:ui] a.ti nr.r,.
The significance of each of those sct.b i: ] is tod Lolov:
. Four-week set Ko.l. compares control
populations of April 5 to hunzc p<.n none
and crirjhlcrophenol populations devel-
oping at the end of the first concen-
tration levels of these cor.ipounds;
Four-week set. No. 2 controls on May 5;
benzoquir.one and TCP at the end. of trie-
second concentration levels; and TCA ncar
the end of the first concentration .level.
Four-week set No.3 - controls on Juno 1;
benzoquinone and TCP approximately one
week- after the third concentration levels
ended; ai.u TCA about mid-way throurh the
second concentration level;
Two-week set No.l '- control-; or. Apr'i?. 20;
bcnzoquincnc and TCP niid-wrr; through the
second concentration levels; and TCA about
nid-wav through the first concentration
level/ " -
Two-week set No.2 controls on May 20;
bonzoquinone -and TCP hear the end of the
third concentration levels; and TCA at the
beginning of the second concentration ^-vel.
"wo-week set No.3 controls on Juno L?;
' and TCA just after the second con con:, rat. ion
level ended.
"':.-.' da^a f i or. tiie microscopic observations :-ro';ran: ar-:- s::r.~ar-
izod i:. F;'-ures 32 through 38. For each pair of parall-.:! ex-
periments, units, four population ccn;::ts were nade curir." -^ach
sv: cf >: iorvatior.s. Tiic maxinum, averar.c, anfi r.inir.ur.-. ro; u] i-
t_cr.5 cf '_ach: set were recorded.
v.r-1." 32 and 33 are hargraphs of t'.-.o averac;o r.icrofa.'.na r-rp-
:i.-:-.s :;: each set of four-week and tw,---v ^ek ohsorvatirv.s, re
v" . iv.-ly, and each population i o broken dov r. ir.tr :Vr cr-npo-
_n-:n. A.: Li-.o-jcsh. the control Copulations ex'r.iiitc'u C'^nri dc-rai .!-
-------
Qc'5
Q.
U
k-J
O
." l-:
;- I-II'MM
:-i l':!:-L..I;:ii; ki
:. r. ' '.-.?.
AVOE3AE
ROr'r:ESS
'; '- : '-, '
5 ? ri-
-1 F3 ! ,
'.-: -H L-.d i
-------
r.OO,
'JOG
1
"_i ULS
I i
j
-------
.
j ,; 350
i O
t
i . *
: . c 300
i ' ;r'
'1
r)'! f \
: " ,
1 ;-:'
1 v; 200
0,
~ ! 50'
L,
O
l"j 1 00
'
50
1 0
i
)
o
^ 300
i
i if.
ti.
co '- -* w
V J
2-V/EEK ,-./,-,i
VCR ;.rAUN'A
_
_
x_ .. _ ..
j ^ .\
i r |
,', l' '?
' J- i
~ ' '
i
i i "
.,__
ST." WO.i
i "
i
! '
f ._ 1.
'." ^ [ - ;
1
^ *"' f1-"
1 "
:
,
i i
J |
\ x -1 '-!*'
'r | "i"
j *A *
i ;
i
' i
I _-.
;
.. _?_ .r:_ . .. _ 1
A " ' ' '
1 ''
SET KO.;--' SH: f'.'J. 3
n
.; . y. r r -
"''" ' VS
'.''
i.' , . i . - 1 -, ~.> f
'r -...-..
\ .. .\
i
,S! r- .'':- !' It" '"..'e
3
i
i
r
(
;
1
j'
1
i
::00 i
so
FIGURE S
TOTAL MICROrAUMA PO.-L'LATiO\!3
-------
350
o
f>
1?. 300
or
Ul
a 250
to
I
-J.
o 200
a
- 150
u.
O
a:
^
11'
50
O
2 -WEEK STALKED
CILIATE POPULATIONS
X A- -
-
1
-
t
MAX. ,
X--I A
>' o A
- f-!--l--^v?
'> i " t
i 1 x i
x ° . -. .- - o A ''-'
o p P y.~
350
CO
(T
250
f? 200 -
1 50
O
ir.
SET NO. 1
SET NC.2
SET ,\0. 3
4-WEEK STALKED
C1LIATE :OPULAT!C\'S
X -CC'N '!«!
0 - i-r :.'(. - .'iM"'.E
n - ' ; I-
A - * C A
_.-!
f
...i
1 !
. FIGURE 3.5
STALKED CiLIATE POPULATIONS
-------
! -V.'E t.H '.'0 r !L 1-
'JO
FIGURE 36
.MOTILE CILiATE POPULATIONS
-------
r'
(X
n"
v>
'
-------
300
:: - -A i " K
.'JV
fi - ','.'r r K
H:.':f -c-:''L IK
BLUE-GREEN /'-.GAE r-Op:jLA'rIO\T
-------
var i.a t.j on fr -m one st'i. Lo another , certain significant devia-
tions occurred in some of the test populations. Most notably,
':.-. it!; the '."our-w-jek and the ^wo-v/eck bonzoquinone populations
.i':.o'-: a Jaic.lane at t'no end of. the second concentration level,
Kit ospeci a] iy duiing and after the third level, due primar-
ily co tl,e dacreas;; in r.talked ciliates. The average four-
wrek control stalked ciliate population reaches a minimum of
ijG ..ndi vidua.'.y per square centimeter or 375 of the microfauna
population, v:hi.lc the bcnzoquinonc population drops cc 5 in-
r"; iV i.duals ur 12';- in observation SCT; No. 3. In a similar fash-
i::ri. the riveracic two-wcok bnnxoquinon.-: stalked ciliate popu-
iau'.oa drc;>3 to 5 individual.'-, or f-*, ir. observation set No.2,
v;hilo Die >:;;trol niinjnvu.n is 50 individuals, or 23%. It would
^ or; Lhuji, u:v?.c l.C'H^oouir.onc has an iniiibitory effect on this
ur>'janisn a L a concerrcra Lion level of 1 mo/1 (concentration r;o.
",';' and po:;r_ijiy at a co'.'.centration arj low as 0.10 mg/1 (concen-
;;:;> tion Xo. ? ) .
\.".-.ij.o the decj-inc in the TCP total r;icrofauna population' is not
,\n''v.arkcd, the TCP stacked ciliate population drops to a mini-
r.uri of 10 ir.uividuals Jn four-v;eek observation set Mo. 3 and
*..fo-v.'c.ek- Gi'.'-:t:rvation dot No. 2, being 12% and 8?> of those mic-
..'orau.-Ki [.emulations--,, rusrcctivcly. Thus, TCP seems to affect
t!:i:-; orcan~sn\ at a concentration of -1 ppni (concentration No. 3).
The 7C.\ ;;u crsfauaa ;'opu] ations compare favorably to the con-
trols .\.li..i:oufjh the :.:our-v:oek stalked ciliate population does
dcc:'ii-.o '.!. .-tt :-.o...J. to 15 individuals or 14%, indicating a
:...-.; .1 In :.:.'. ibilorv effect at the second concentration level
! 1
roiiira:-'. to the stalked ciliates, the control motile cili-
popuLu .:.;;:, in quite; stable over the course of the bioassays.
\.*iva i'-.r.tnnco oi rigr.i f icant deviation in the te?t motile
.a1-.-a p'.;'.;la.Lions occurs. The four-week ben;coquinone popula-
. J.uclJnos to a ini:;ir.-.un average of 10 individuals per square
i jb.se rva L i en :;et ',',0 . Z , as compaired to a control
c.'.~ 3'J .i nai"iduals, indicating a possible ir.hib-
of tho tliird concentration level.
roti"i.c:!i, and nesnatoda populations flue-
.;ree and v:cre so small in comparison
..', a:"- -t;o i.iake evaluation of possible
. = :v.i i'~l :']->',. t':.r- riax.i mum, average, and minimum
;:i:cn ;.ja:.a, nt:alj'.vd ciliatf^s, and mcti]e ciJ.i-
clv, f'-.':. : i I. c/usi. rva tior. sets. ^.'ote' the dt^cline
'.:,d r.f.il.:.-_d cilirte populations ir. the TCP and
-;;L unit-, .ir.r. tro c-.-ar.ural stability of tlv= mo-
; .; 1.1 tioj:tr. -"Mot .n. for benzoquitione ir. t'our-v/ee}:
, "!-...:'. '.':: data, on micro^aur.a populations woul'1
-------
seen to- indicate inhibitory effects of the compounds, espe-
cially with regard to stalked ciliates, at the follov:ing nom-
inal concentration levels:
benzoquinone at 1 ppm (concentration 3)
. and-possibly 1/10 ppm (concentration 2);
TCP at 4 ppm (concentracion 3); and TCA
at 1.. 4 ppm (concentration 2).
There appeared to be no effects on the microfauna of any of
the compounds at lower concentration levels.
Four categories of microflora were observed in the experimen-
tal aquaria: diatoms, blue-green algae, green algae, and yel-
low-brown algae. Possible inhibitory effects were seen on two
of these, namely, the diatom populations and the blue-green
algae populations.
Figure 37 plots the maximum, average, and minimum diatom pop-
ulations in Hi.1 observation sets. The four-week control pop-
ulations wori'- exceptionally stable throughout the duration of
the bioassays, ranging from 17,000 to 65,000 cells por square
centimeter with an average of 47,000. The fcur-v;eek benzoqui-
none populations exhibited a steady decline, finally deviating
significantly in observation set No.3, with an average of;only
9,000 cells and a maximum of 13,000. TCP exhibits a similar
trend, declining to an average of 8,000 and a maximum of -10,000.
TCA diatom populations remained somev.hat higher than the control
populations in both sets of four-week data.
The two-week control diatom populations were also stable, with
a maximum of- 40,000, a minimum of 9,COO, and an average of
25,000 cells per square centimeter. In observation set No. 2 ,
both the benzoquinonc and the TCP diatom populations averaged
only 5,000 cells with maximum of 7,000. The TCA populations
were again somewhat, higher than r.he controls, except in set
No. 3, v/iir.h an average and a maximum of 9,000 and 18,000, re-
spectively.
Thus, there is inconclusive evidence of inhibiuory effects
on diatoms of benzoquinone at 1 ppr.i (concentration 3), and
possibly 1/10 ppm (concentration 2.) , TCP at 4 ppm (concentra-
tion 3), and TCA at 1.4 ppm (concentration 2).
The cont\ol populations of blue-green algae, as evident in
Figure .'S, wore extremely variable, reaching a maximun of
more than one million ceils and a minimum of 25,000 cellb in
four-week populations. The most significant observation
lies in the fact that, while tremendous blue-arcen algal
blooms occurred in the controls, in general ;he populations
remained 3ow in the test units. . It is not known, however, if
this was due to the action of the compounds, or to sono phys-
ical vac,ary in the controls at the time of the blooms.
- 98 -
-------
In summary, the data fror. the rr.i rror'-o: i : '; :;". v: "- i~~'.r.
indicate tiiat all t.l.rc': ro;npoui,t.:-j, !_:-; xi. .. : .A :.-i:n".n*;u.. :.<;: .'.;;id
TCP, may possibly inhibit diatoms and stalked ciliates at the
highest concentration levels of tno compounds. Additional
evidence suggests that benzoquinone rany inhibit those rdcroor-
ganisms at its second concentration level. The variations in
basic population in the control unit were significant, and a
detailed statistical examination of the population variance
would be necessary to quantitatively define the impact of the
various test compounds. ?o carry out such a study would re-
quire the collection of a large number of observ it i r "is, nn.:
would be beyond economic practicality.
Summary
Substantial insight into the validity of the intermediate-term,
flow-through bioassay as a method for determining toxicity was
gained. Laboratory facilities incorporating model freshwater
ecosystems in a flow-through system v:ere designed. Methods
for testing compounds and monitoring the ecosystems' response
to compounds were developed. Three chlorinated organic com-
pounds were investigated for toxicity to a variety of aquatic
test organisms.
It was found 'that the te.c,t fish, pimephales pronie las, was able
to tolerate higher concentrations of p-bcnzoquinone and 2,4,6-
trichlorophenoi in the flow-through studies than in the four-
day static bioassays. As a result of its limited solubility
in water, 2,4,6-trichioroaniline could not be dispensed to the
model ecosvstems at a concentration level as high as that de-
termined to be lethal in the static tests. The variation ir
control fish populations during the flow-through bioassays
mace it impossible to determine sub-lethal effects of the
compounds.
Qualitative analysis of Kiicrocrcv.n.ism population data suctnested
inhibitory or toxic effects or. diatoms and stalked cilia'.'es of
the three compounds at the highest concentration levels tested.
However, the instability of the control microorganism popula-
tions prevented valid statistical treatment of the dntr. A
great increase in effort, well above that practical for a pro-
ject of this nature, would be necessary before strict defini-
tions of toxocity to microorganisms could be determined.
Of the three species of rooted vascular plants tested, only
Lud v i g i a exhibited an interpretab.lo growth pattern. Compari-
sons between test and control plants indicated no toxicity of
any of the compounds tested. N'o a.".vc-::so effects on the out-
ward appearance of any of the three species ,:::« observed.
The experimental ecosyr,tc~s wero unsuitable for the mainten-
ance of stable macroinvcrtebrate populations for a duration
sufficientlv lone; fcr cor.i:-,ound testing.
-------
Although considerable information of a qualitative natr.re was
gathered, the flow-through studies did not yield reliable
quantitative data from which conclusive determination:: of toxi-
city could be made.
Considering the relative efforts involved, the intermediate-
term, flow-through bioassay did not compare favorably to the
routine static bioassay as a reliable experimental method.
- 100 -
-------
SECTION XII
' ACKNOWLEDGEMENTS
This project was undertaken by Hydroscience, Inc., under the
direction of Edwin L. barnhart. The experimental studies
completed dur:ng the various phases of the project were con-
ducted under the immediate supervision of George P.. Campbell,
Dr. George J. Kehrber
-------
SI,JTION XIII
REFERENCES
1. Ingols, R.S., and Jacobs. G.M., "BOD Reduction by Chlor-
ination of Phenol and Amino Acids," Sewage and Industrial
Wastes, 29, No.3, pp.258-262 (1957).*
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pp.1517-1527 (1963).
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'! Jour. V\F.r.C.F., _3_1 / Mo.4, pp.629-635 (1966).
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and Physics, 42nd Edition, The Chemical Rubber Publishing
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6'. Roberts, J.D., and Caserio, M.C., Basic Prinicples of_ Or-
i ganic Chemistry, W.A. Benjamin Company," New York"] (T965T.
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(1951).
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Phenolic Tastes by Chloro-Oxidatior." , Water and Sewage
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Phenolic Compounds", in Principles -and Applications of
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13. Marks, H., by private conimunication.
Preceding page blank
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s I-::.!-: en-: it WATF.X
.'<.':souKCt-:s A KSTRA crs
!.\i'UT TRANSACTION FORM
.(. .-ti:i>-.<>iu.i No.
I Vi.-.V .1.
F.FF7.CY OF Ci::,ORINv.TION Oh SELECTED ORGAMC CHEMICALS <:.
'. /V'lViT A'o.
12C20 EXr,
i A'o.
iiiydrosciur.ee. Inc., Westvood, ;;ew Jer.-.ey 07675
j
j'5. .S'::;i;>i'c:;:('/.r.!/y A'o:*'.s
Subrnitl.od vc v.;ic V.'ater O'aclicy Off-Leo, Flnvironr. or.lT. j i-'roi.oction /.cency.
L-'ourtccn i::c;-jstriai orgcnic ch-nicnls v.-cro cxonincd for their persistence i
t:;rou.;;h ;-:.olooica 1 tro.~ t:\er.t, cither r.s ::ia initial cor.pcur.ds, or cis !
cic^'fculat.ic.:1: j-rociucts. Sc::!i-ccr.l.i:-.uci!S aotivr:ccd sludge syctc.r.s v.'crc C.T.- j
ployed. The ability o;" 0,20:1 o:" t'ro ci'.cr.lcalc to participate in reactions i
v.-itii Lrrec chlorine v;as then dotorir.inea in a series of bate:-, experiments. i
It v.\!is found th;.t certain of the tost co~pou;:dr, forr.od persistent dccrada-
'cior. ;>rc.:.ucts riiiring troatir.ent. Five oi" the initial compounds reacted
|rcnt:ily with chlorine, ur.der con-litior-s cor.rr.or.iy cir.ployoci in effluent
c:hlori.ici'.:i.o:i.
Five of t'c chlorir.aticn products '..-ore further studied in respiror.ctsr ex-
>orir.'.ent.s to evaluate tiicir porsistonce in :r.i:-.ec! iTiicrohiai systems, 7'r.eir!
it-Jxioity to fish was doterr-incd v.cinc; the stat-i.c hioassay procedure. j
;Ir. w/iO final phase of -ci'.e study, a soric-s of bench scale, continuous flow j
_cosys".o~,s \:ore established for the evaluacior. of /.oncer tci'- effects o. |
th::>.:o of che chlcrination products. Several varieties of organises, rep- j
jrescnting different levels in the food chain, were studied.
*C:ilorin^.tion, *Cher.iicai Reactions, *Diodograriation, *3io?.ssay,
*Scosyster.ir., *Toxici ty
0rr;r.nic Che::.ic;-.lo , *. \ctiva ted Sludoe, ^Detjr-idatior. Products,
uces, *Respiro::.eter Studios, *"or.':inrous Flow
. .- .-I..U;! ^ - -, -> - - ,- -
v ^ I?. 0^.-., u'j
.,^^:.:JrIo^ ::\'drO3CJ Cr.CC , Ir.C.
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