ARCHIVES HISTORIQUES
DELACOMMISSION
COLLECTION RELIEEDES
DOCUMENTS "COM"
COM(80)40
Vol.1980/0016

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COMMISSION OFTHEEUROPEAN COMMUNITIES
PROPOSAL FORACOUNCIL DIRECTIVE
ONTHEAPPROXIMATION OFTHELAWSOFTHEMEMBER STATES
RELATING TOMETHODS OFTESTING THEBIODEGRADABILITY
OFNON-IONIC SURFACTANTS ANDAMENDING
DIRECTIVE 73/404/EEC
(presented bytheCommission totheCouncil)COM(80)40 final
Brussels/8thFebruary 1980
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C0MC80)40final

1.This proposal for aCouncil Directive comes under the General Frogra ;'.5Tie
for theremoval oftechnical barriers totrade ,which theCouncil
approved on28May 1969 ,inline with which twoDirectives relating to
diesame sector have already been adopted .These are the Council
Directive of22November 1973 relating todetergents andtheCouncil
Directive ofthesame date relating tomethods oftesting the
biodegradability ofanionic surfactants (1).
2.Themeasurement ofthelevel ofbiodegradability bydifferent methods
.can cause distortion ofthe Community market andhave adirect effect
onitsoperation .ACommunity solution proposing asingle method is
therefore necessary .
3.Thebiodegradability ofdetergents isone aspect ofthemuch more wide-
ranging andvery topical problem ofenvironmental pollution ingeneral
andwater pollution inparticular .
Forthese reasons industry ,inagreement with public authorities ,has
progressively shifted itsproduction todetergents containing
biodegradable surfactants .Nevertheless ,'forcertain applications in
the food industry ,themetallurgical industries and indishwasher
detergents ,small amounts ofcertain non-ionic surfactants oflow
biodegradability must beused fortechnical reasons :machines used in
these industries work athigh speed andhave touselow-foaming and/or
highly alkali-resistant surfactants ofhigh detergency .For thetime
being there isnosurfactant which is80%biodegradable andhasthese
properties .
The amounts used inthis way are small and thetotal EECconsumption is
estimated atsome 5500-8500tonnes peryear ,i.e. ,less than 1%
ofallthesurfactants used .incleaning products .
Itshould also bepointed out that these surfactants ,although oflow
biodegradability ,arenot asfar asitisknown responsible for
ecological problems inthe'Community andthat ,even ifthey were ,such
problems would bestrictly local ..
Research isinprogress inseveral Member States with aview to
developing satisfactory substitute products which aresufficiently
biodegradable tocomply with theDirective .
(1)OJNoL347 ,17December 1973 ,pp.51and53
t
•/.

4.Technical progress necessitates aprompt adaptation ofthetechnical
standards' inthemethods fortesting thebiodegradability of
surfactants .Tnat isthereason why this Directive provides for the
institution ofaprocedure ofclose collaboration between theMember
States and the Commission within the framework ofthe Committee for the
•adaptation totechnical progress ofthedirectives relating tothe .
removal oftechnical barriers totrade indetergents .This procedure
istheoneprovided forbytheResolution of28Kay1969 andsubsequently
adopted bytheCouncil indifferent directives .
5.Inaccordance with theOpinions which theEuropean Parliament andthe
Economic andSocial Committee delivered onthebasic "Detergents "
Directive ,thesolution of "total "harmonization hasbeen chosen .
II.COMMENTS ONSPECIFIC ARTICLES
Article 1
This Article establishes the scope oftheDirective .
Article 2
Article 2stipulates that adetergent must notbeplaced onthemarket if
itisdetermined that thelevel ofbiodegradability ofthenon-ionic
surfactants itcontains isless than 80%,andprovides that themethods
used forthis determination ■ -i1bethose oftheOECD ,theFederal
Republic ofGermany ,France and the United Kingdom .
Article 3
This Article lays down theprocedure tobefollowed intheevent ofa
dispute .
Article 4
This gives atemporary exemption forcertain uses ofnon-ionic surfactants
with abiodegradability ofless than 80% .
Article 5 '
This Article provides for aprocedure forthe adaptation totechnical
progress ofthedirective concerning detergents .
Articles -6and 7
These Articles are common toall directives .- '
III .CONSULTATION OFTHE SECTORS CONCERNED
When preparing this Directive ,the •Commission took note ofthecomments made
bytherepresentatives ofthe industries concerned .,
IV*CONSULTATION 'OFTHEEUROPEAN PARLIAMENT ANDTMEECONOMIC ANDSOCIAL
COMMITTEE
Inaccordance with thesecond paragraph ofArticle 100 ,theopinion ofthese
twoinstitutions isnecessary . ' .

ontheapproximation oftheiaws-of theMember Stated -
relating tomethods oftesting thebiodegradability
ofnon-ionic surfactants andamending Directive 73/404/EEC ''
THE COUNCIL OFTHE EUROPEAN COMMUNITIES ,
Having regard totheTreaty establishing theEuropean Economic Community ,and
in .particular Article 100thereof ; . -
Having regard to .theproposal ..from theCommission ;
«
Having regard totheopinion oftheEuropean Parliament ;
Having regard tothe0/pinio.n oftheEconomic andSocial Committee ;-
Whereas themethods ,oftesting inforce intheMember States ,while pursuing
thesame objective ,differ incertain respects and are thus detrimental to
thefunctioning ofthecommon market ; ...
1 .Whereas Council Directive 73/404/EEC relating to
*
detergents provides in .Article 4fortheadoption ofdirectives
specifying *methods oftesting inorder toascertain that therequirements
ofthat Directive arebeing observed ;whereas Council Directive 73/405/EECof
222November 1973 relating tomethods oftesting thebiodegradability of
anionic surfactants specified such methods andtolerances foranionic
surfactants ; '
Whereas ,toenable -Member States todetermine thelevel ofbiodegradability
ofnon-ionic surfactants ,themethods oftesting ,already inuseforthis
purpose incertain Member States maybeused ;whereas ,however ,inthe
event ofdisputes ,biodegradabi lity should betested -byacommon
reference method ;
OJNoL347,
^ •
2OJNoL347,"17.12.1973 ,p.51
17.12.1973 ,u.53

Whereas inregard tothe a^pro^ir .ationofthe lav:s of Memer States
relating todetergents ,suitable tolerances for meas'arinj biodegr adabil ity
should bedetermined ,asprovided for in-Article 4ofCouncil Directive
73/404/EEC inorder totake account ofthe
unreliability oftest methods which could lead torejection decisions with
onsirerable cccno -icconsequences ;whereas arejection decision must betaken
only iftheresults obtained byananalytical method mentioned in
Article 2show alevel ofbiodegradabiiity lover than 80% ;
T.vhereas forthe time being small quantities ofcertain non-ionic surfactants
oflowbiodegradabiiity must beused forsome purposes because oftechnical
problems andinorder toprevent other undesirable effects onhealth and
theenvironment ;Whereas itwill nevertheless benecessary toreview theuse
ofthese surfactants oflowbiodegradabiiity inthelight oftechnical
progress ;
Whereas technical progress necessitates arapid adaptation ofthe technical
requirements specified bytheDirectives ondetergents ;whereas ,to
facilitate theintroduction ofthenecessary measures ,asystem should be
setupproviding forclose collaboration between theMember States andthe
Commission bymeans ofaCommittee fortheadaptation ,inthelight of
technical progress ,oftheDirectives ontheremoval oftechnical barriers
totrade indetergents ,
HAS ADOPTED THIS DIRECTIVE :
Article 1
This Directive concerns themethods oftesting tinebiodegr adabil ityof
non-ionic surfactants used indetergents .

.r,accordance with theprovisions ofArticle 4of Directive
'3/404/EEC , dueaccount being taken oftheunrei' l -iit ■
>•:testing methods ,theMember States shall prohibit theplacing onthe
market anduseontheir territory ofadetergent ifthelevel of
blodegr adabil ityofthenon-ionic surfactants contained insuch
detergent isless than 80%determined inaccordance with oneofthe
Coilowing methods :
-TheOECD method ,published intheOECD technical report of11June 1976 or»
the "Proposed Method fortheDetermination oftheBiodegradabi lity of
Surfactants used inSynthetic Detergents ";
-Themethod inuse intheFederal Republic ofGermany ,established bythe
"Verordnung uber die Abbaubarkeit anionischer undnichtanionischer grenz -
f1achenakti ver Stoffe inWasch -undReinigungsmitteln -"of30January 1977 ,
published intheBundesgesetzblatt ,Part I,page 244 ;
-Themethod inuse inFrance ,approved bydecree of28December 1977
published inthe "Journal Officiel delaRepublique Frangaise "of18Januar
1978 ,and experimental standard T73-270 March 1974 ,published bythe
"Association frangaise denormalisation "(AFNOR );
-Themethod inuse inthe United Kingdom called the "Porous Pot Test "and
described in Technical Report 78 (1977 )bytheWater Research Centre .
Article 3
Under theprocedure laid down inArticle 5(2)of
Directive 73/404/EEC, thelaboratory opinion on .non-ionic
surfactants shall bebased onthe "Confirmatory test procedure "described
intheAnnex .
.•/.

-6~
Article 4
Th?following Articles shall beinserted inDirective 73/404/EEC :
Article 2a
1.Until 31December 1984 ,Article 2shall notbeapplicable tothe
following non-ionic surfactants :
a)lowfoaming polyalkene oxide addition products used inindustrial
cleaning agents andmechanical dish-washing products ;
b)alkali-resistant terminally-blocked alkyl andalkyl-aryl polyglycol
ethers used inindustrial cleaning agents forthefood ,beverage and
metal working industries .
2.During that period ofexemption ,theexemption shall bere-examined and
where appropriate adapted inthe light oftechnical progress inaccordance
with theprocedure laid down inArticle 7b.
3.Intheevent ofindustrial useofthesurfactants referred toinparagrah 1,
theuser shall employ thebest techniques available inorder tolimit the
emission ofthese surfactants into theaquatic environment .
"Article 7a
1.ACommittee shall beestablished fortheadaptation totechnical progress
ofDirectives forremoving technical barriers totrade inthedetergents
sector ,hereinafter called 'the Committee' ,which shall consist of
representatives oftheMember States under thechairmanship ofa
representative oftheCommission .
2.TheCommittee shall establish itsrules ofprocedure .
•/.

ι
••
"Article 7b
1.Where -recourse ishadtotheprocedure definedinthisiftrticle,the
r
mattershallbereferred totheCommittee byitschairman,onhis.
owninitiative orattherequestoftherepresentative ofaMemberState..
2.TheCommission representative shallsubmittotheCommittee adraft
ofthemeasures tobetaken.TheCommittee .-shalIdeliveritsopinion
onthedraftwithin aperiodwhichmaybefixedbythechairman
according totheurgencyofthematter..Amajority oftwelvevotes
shallberequired beforeitcandeliveritsopinion,thevotesofthe
MemberStatesbeingweighted aslaiddowninArticle 148(2)ofthe ■
Treaty.
Thechairman shallnotvote.
3..(a)TheCommission shalladopttheproposed measures wheretheyare
inaccordance withtheopinionoftheCommittee .
(b)Wheretheproposed measures arenotinaccordance witjitheopinion
oftheCommittee ,orifnoopinionisdelivered ,theCommission
shallsubmittotheCouncilwithoutdelayaproposal onthe
measures tobeadopted. ~~ ,.'
,(c)If.theCouncil hasnotactedwithinthreemonthsofthedateon
whichtheproposal wassubmitted,theproposed measures shallbe
adopted bytheCommission .
Article 7c
Theappropriate methodsoftesting andtolerances ,definedeither
directly orbyreference tostandards laiddownoutsidetheinstitutional
framework oftheCommunity,inthe.Directivesreferred toinArticle 4
shallbemodified,inparticular inordertoadaptthemtotechnical
progress inaccordance withtheprocedure laiddowninArticle 7b."

„8_
Article 5
1.TheMember States shall bring into force theprovisions necessary to
comply with this Directive within aperiod oftwelve months following its
notification .They shall forthwith inform theCommission thereof .
2.Member States shall communicate totheCommission thetext oftheprovision
ofnational lawwhich they adopt inthefield covered bythis Directive .
Article 6
This Directive isaddressed tothe Member States .

ANNEX
'DETERMINATION OFTHE BIODEGRADABILITY OFNON-IONIC SURFACTANTS
REFERENCE METHOD
(CONFIRMATORY TEST )
CHAPTER 1
1.1 . Definition
Non-ionic surface active agents inthesense ofthis directive
are those surface active agents ,which after passage through
cationic andanionic ionexchangers aredetermined asbismuth
active subs tance '(BiAS )according totheanalytical procedure
described inchapter 3.'
1.2 .Equipment needed formeasurement
Themethod ofmeasurement employs thesmall activated sludge plant
shown infigure 1,and ingreater detail infigure 2. '
Theequipment consists ofastorage vessel Aforsynthetic sewage ,
dosing pump B,aeration' vessel C,settling vessel D,air lift pump E
torecycle theactivated sludge ,andvessel Fforcollecting the
treated effluent .
Vessels AandFmust beofglass vorsuitable plastic andhold at
least 24litres .Pump Bmust provide aconstant flow ofsynthetic
sewage totheaeration vessel ;this vessel ,during normal operation
contains 3litres ofmixed liquor .Asintered aeration cube G
issuspended inthevessel Cattheapex ofthecone .Thequantity ,
ofairblown through theaerator should bemonitored bymeans ofa
flowmeter H.
1.3 .Synthetic sewage
Asynthetic sewage isemployed forthetest .Dissolve ,ineach litre
oftapwater : ..
•S '
160mgpeptone
110mgmeat extract - .
30mgurea
7mgsodium chloride
Amgcalcium chloride ,2^0
2mgmagne sium sulphate ,7HO ,
28mgofdipotassium phosphate (K_HP0 ,)
and 10+1mgBiAS . -
• , _ *-
The ;BiAS isextracted from theproduct 'tobetested bythemethod
given inChapter 2.Thesynthetic sewage isfreshly prepared daily .

η·2^
.1.4,Preparation ofsamples
1.4.1 .Uncompounded surfactants may beexamined intheoriginal
state . The BiAS content must bedetermined inorder
toprepare thesynthetic sewage (1.3.).
1.4.2 .Formulated products are analysed for BiAS ,MBAS and
soap content . They must besubjected toanalcoholic
extraction and to .aseparation ofthe BiAS (see '
Chapter 2).
The BiAS content oftheextract must beknown inorder toprepare
.the synthetic sewage .
1.5 .Operation ofequipment
Initially ,,fill aeration vessel Candsettling vessel Dwith
synthetic sewage . The height ofthevessel Dshould beso
fixed that the volume contained inthe aeration vessel Cis
3litres . Inoculation ismade byintroducing 3mlofa
secondary effluent ofgood quality ,freshly collected from a
treatment plant dealing with apredominantly domestic sewage .
Theeffluent must bekept under aerobic conditions intheperiod
between sampling and application . Then set the aerator G,
airlift Eand dosing device Binoperation . The synthetic
sewage must pass through theaeration vessel Catarate ofone
litre per hour ;this gives amean retention time of3hours .
The rate ofaeration should besoregulated that the contents of
vessel Care kept constantly insuspension and the dissolved
oxygen content isatleast 2hig/1. Foaming must beprevented
byappropriate means .Antifoaming agents which inhibit the
activated sludge orcontain BiAS must not beused . Theair­lift pump E-must beset sothat theactivated sludge from the
settling vessel iscontinually and regularly recycled to
aeration vessel C. Sludge which has accumulated around the top
oftheaeration vessel C,inthe base ofthesettling vessel D,
orinthe circulation circuit must bereturned tothe circulation
atleast once each day bybrushing orsome other appropriate
means . When the sludge fails tosettle ,itsdensity may be
increased bytheaddition of2mlportions ofa5per cent solution
offerric chloride ,repeated asnecessary .
Theefflent from thesettling vessel Disaccumulated invessel F
for 24hours ,following which asample istaken after thorough
mixing .Vessel Fmust becarefully cleaned ,,

1.6 .Checking measuring equipment
,TheBiAS content (inmg/1)ofthesynthetic sewage isdetenn.ned
immediately before use .
The ?iAScontent (inmg/1)oftheeffluent collected over 24
hours invessel Fshould bedetermined analytically bythesame
method ,immediately after collection :otherwise thesamples must ,
bepreserved ,preferably byfreezing .Theconcentrations must
bedetermined tothenearest 0.1mgBiAS /1. -
Asacheck ontheefficiency oftheprocess ,thechemical oxygen
demand (COD)orthedissolved organic carbon (DOC)ofthe
filtered effluent accumulated invessel Fand ofthe filtered
synthetic sewage invessel Aismeasured atleast twice perweek .
rThereduction inCODorDOC should level offwhen aroughly
.regular daily BiAS degradation isobtained i.e. attheendofthe
running-in period shown "inFigure 3..
The loss ,onignition ofthedrymatter intheactivated sludge
intheaeration tank should bedetermined twice aweek ing/1.
•Ifitismore than 2.5 g/1,theexcess activated sludge must be
discarded . ;
Thedegradation test isperformed atroom temperature which should
besteady andkept between 291and298KC18-25°C ).
1*7 .Calculation ofbiodegradability
Thepercentage degradation t>fBiAS must becalculated every day
onthebasis oftheBiAS content ?inmg/1ofthesynthetic sewage
andofthecorresponding effluent accumulated invessel F.
The*degradability figures thus obtained should bepresented ,.
graphically asinFigure 3. " .
Degradability oftheBiAS should becalculated asthearithmetic
mean ofthefigures obtained over the21days which follow the
running inperiod ,during which degradation hasbeen regular
andoperation oftheplant trouble –free . -
Inanycase theduration oftherunning-in period should not
exceed sixweeks .
Thedaily degradation values arecalculated tothenearest
0.1percent biitthefinal result is .given tothenearest whole
number .
Insome cases itmaybepermissible toreduce thefrequency of
sampling butatleast 14results collected over the21days
which follow therunning-in period should beused incalculating
theaverage . . .
.7.

^L-
CHAPTER 2
PRELIMINARY TREATMENT OF PRODUCTS
TO BE TESTED
2-.1 . Preliminary notes
2.1.1 .Treatment ofsamples
The treatment ofnon-ionic surface active agents and formulated
detergents prior tothe determination ofbiodegradatility
inthe confirmatory test is:'
The purpose ofthealcoholic extraction istoeliminate the
insoluble and inorganic ingredients ofthe commercial
product ,which insome circumstances might upset the
degradation test .
Isolation and separation ofnon-ionic surface active agents
from soap ,anionic and cationic surfactants isrequired
for correct biodegradation tests .
This isachieved byanion exchange technique using amacro­
porous exchange resin and suitable elutants for' fractional
elution . Thus soap ,anionic andnon-ionic surfactants may
beisolated inone procedure .PRODUCTS TREATMENT
Non-ionic surfactants
Formulated detergentsNone
alcoholic extraction
followed by separation ofthe
non-ionic surfactants byion
exchange
2»1#2 .Ion Exchange procédure

-5"
' 2.1.3 .Analytical control
After homogenizing ,theconcentration ofanionic and non-ionic
surfactants inthesynthetic detergent isdetermined according
tothe MBAS and BiAS analytical procedure . The soap content
isdetermined byasuitable ,analytical method .
This analysis oftheproduct isnecessary tocalculate the
.quantities required toprepare fractions for thebiodegradability
tests .
Quantitative extraction isnot necessary ;however atleast
80per cent ofthesurfactants nim-Toni'c should beextracted .
Usually ,90per cent and more isobtained .
2.2.Principle
From anhomogeneous sample (powders ,pastes ,and dried liquids )an
ethanol extract isobtained which contains thesurfactants ,soap and
other alcohol-soluble constituents ofthesynthetic detergent sample .
Theethanol extract isevaporated todryness ,dissolved inanisopropanol /
water mixture and thesolution obtained ispassed through astrongly
acidic cation exchange /macro-porous anion exchange combination heated
to323 K(50 C). This temperature isnecessary toprevent precipitation
offatty acids inacidic media .
The non-ionic surfactants are obtained from theeffluent byevaporation .
Cationic surfactants ,which might upset thedegradation test and the
analytical procedure areeliminated *bythecation exchanger placed
ontop ofthe anion exchanger .
2.3.Chemicals and equipment , '
2.3.1 .Deionised water . ^ -
2.3.2 .Ethanol ,95vol.%C^H^-OH
(permissible dénaturant :methyl ethyl ketone or
methanol ).

-6τ
£.3.3 ,Isopropanol /water -mixture (50/50v/v);
50parts byvolume isopropanol (CH^CHOH .CH^)and50parts byvolume water (2.3.1 )
2,3.4 .Armonium bicarbonate solution (60/40):
0,3 MNH^HC0 oin1000 mlofanisopropanol /watermixture consisting of60parts byvolume isopropanol and
40parts byvolume water (2.3.1 )
2.3.5 *.Cation exchanger (KAT ),strongly acidic ,resistant to
alcohol (50-100 mesh ).
2.3.6 .Anion exchanger (AAT ),macro-porous ,Merck Lewatit
MP7080 (70-150 mesh )orequivalent .
2.3.7 .Hydrochloric acid ,10% HC1 w/w.
2.3.Q %000 mlround-bottomed flask with ground glass stopper
and reflux condenser .
2.3 .9*90mmdia .suction filter (heatable )for paper filters .
2.3.10 .2000 mlfilter flask .
2.3.11 .Exchange columns with heating jacket and cock :
Inner tube 60mmindiameter and 450 mminheight
(Fig .4).
2.3.12 »Water-bath .
2.3.13 .Vacuum drying oven .
2.3.1 /t.Thermostat .
2.3.15 .Rotary evaporator .
1

2.4.Preparation ofextract and separation ofrion-ionic active agents
2.4.1 .Preparation ofextract x
Thequantity ofsurface active agents necessary for the degradation
test isabout 25gBiAS .
Inpreparing extracts for the degradation tests ,thequantity of
'„. product tobeu^ed should belimited toamax .1000 g.Therefore ,
itmay benecessary tocarry out the operation twice inorder
toobtain sufficient quantity for the degradation tests .
Experience has shown that there are advantages inusing a
number ofsmall extractions rather than' one large extraction ..
2.4.2 .Isolation ofalcohol-soluble constituents
Add 250 .gofthesynthetic detergent tobeanalyzed to ■%.
1250 mle.thanol andheat themixture toboiling point andreflux
for 1hour with stirring . Pass thehot alcoholic solution
through acoarse-pored suction filter heated to323 K(50°C )
and suck off sharply ,. Wash theflask and suction filter with
approx .200 mlhot e.thanol . Collect thefiltrate andfilter
washings inafilter flask .
Inthe case ofpastes orliquid products tobeanalysed ,make
sure that not more than 25g'anionic surfactants and 35gsoap are
contained inthe sample .Evaporate this weighed sample todryness .
Dissolve theresidue in 5*00 mlethanol andproceed asdescribed
above .
Incase "ofpowders oflowapparent density (300 g/l)iti6re- ■
commended toincrease the ethanol ratio inthe relation 20.1 .
Evaporate theethanolic filtrate tocomplete dryness,' preferably
"bymeans ofrotary evaporator .Repeat theoperation ,ifagreater
quantity ofextract isrequired .Dissolve alltheresidue in5*000 ml
isopropajiol water/mijtture . , , .

τ8·
2.4.3 .Preparation ofion exchange columns
Cation exchange column
Place 600 mlcation exchange resin ina3000 mlbeaker
and cover byadding 2000 mlhydrochloric acid . Allow
tostand "for atleast 2hours stirring occasionally .
Decant the acid and transfer the resin into the column
(2.3.11 .)bymeans ofdeionised water . The column
should contain aglass wool plug . Wash the column with
deionised water atarate of10-30ml/min until the
eluate isfree ofchloride . Displace the water with
2000 mlisopropanol /water mixture (2.3.3 .)atarate of
10-30 ml/min . The exchange column isnow ready for
operation .
Anion exchange column
Place 600 mlanion exchange resin inabeaker and cover
byadding 2000 mldeionised water .Allow theexchanger
toswell for atleast 2hours . Transfer the resin into the
column bymeans ofdeionised water . The column should
contain aglass wool plug .
Wash the column with 0.3 Mammonium bicarbonate solution (2.3.4 )
until free ofchloride . This requires about 5000 ml
solution . Wash again with 2000 mldeionised water .
Displace thewater with 2000 mlisopropanol /water mixture
(2.3.3 )atarate of10-30ml/min . The exchange column
isnow inthe OHform and ready for operation .
V
2'.4.4.Ion exchange procédure'
Connect the exchange columns sothat the cation exchange
column isplaced ontop ofthe anion exchange column .
Heat the exchange columns to323 K(50 C)using athermostat .
Heat 5000 mlofthe solution obtained initem 2.4.2 to333 K
(60c)and pass thesolution through the exchanger combination
atarate of20 -ml/min . Wash the columns with 1000 ml
hot isopropanol /water mixture /i.3.'i ).

~"s**
Toobtain thenon-ionic surface active agents ,collect
thefiltrate andfilter vashings and evaporate todryness ,
preferably bymeans ofarotary evaporator . Theresidue -.contains the BiAS . Add deionised water until adefined
volume isobtained and determine the BiAS content asinitem
3.3 inanaliquot . The solution isused asastandard solution
ofnon-ionic surf actantfi -for the -degradation test . The solution
should bekept atatemperature below 278 K(5 C). »
£.4.5 .Regeneration ofexchange resins v _
The cation exchanger isrejected after use .
The anion exchange resin isregenerated bypassing about
5000-6000 mlofammonium bicarbonate solution (2.3.4 )
down the column ataflow rate ofapproximately 10ml/min .
until theeluate isfree from anionics (methylene blue test).
Then pass 2000 mlisopropanol /water mixture (2.3.3 )down
the anion exchanger towash . The anion exchanger isagain
ready for operation . ,•

--10-
CHAPTER 3
•DETERMINATION OF NON-IONIC SURFACE
ACTIVE AGENTS IN BIODE GRADAT ION TEST LIQUORS
Introduction
Surface active agents are concentrated and isolated bygas
stripping . Inthe sample used ,the quantity ofnon-ionic
surfactant should beintherange 250-800 /Jg.
The stripped surfactant isdissolved inethyl acetate .
After phase separation and evaporation ofthesolvent ,the
non-ionic surfactant isprecipitated inaqueous solution with
modified Dragendorff reagent (KBil ,+Bad -+glacial afcetic
acid ).
Theprecipitate isfiltered ,washed with glacial acetic acid
and dissolved inammonium tartrate solution . The bismuth
inthe solution istitrated potentiometrically with pyrrolidinedi -
thiocarbamate solution atpHv4-5 using abright platinum indicator
electrode and acalomel orsilver /silver chloride reference
electrode .
The method isapplicable tonon-ionic surfactants containing
6-30 alkylene oxide groups .
The titration result ismultiplied bytheempirical factor
of54for conversion tothereference substance -nonylphenol
condensed with 10mols ethylene oxide (NPlO ).

Reagents and equipment
Allreagents are tobemade upindeionised water .
3.2.1 .Pure ethyl acetate ,freshly distilled .
3.2.2 .Sodium bicarbonate NaHCO .A.R.
3.2.3 .Dilute HC1 (20mlhydrochloric acid A.R. conc .per 1000 mlwater )
3.2.4 .Methanol A.R.,freshly distilled ,kept inaglass bottle .
3.2.5 .Bromocresol purple ,0,1 gin100 mlmethanol .
3.2.6 .Precipitating agent ":theprecipitating agent isamixture •
.of 2volumes ofsolution Aand 1volume ofsolution B. ^
The mixture isstored inabrown bottle and can beused upto
one week after mixing .
3.2.6.1 .Solution A
Dissolve 1.7 gbismuth (III)nitrate A.R.,(BiO.NO^.H^O )in20mlglacial acetic acid ,and make upto100 ml
with water . Then dissolve 65gpotassium iodide A.R. ,
in200 mlwater . Mix these two solutions ina1000 ml
- measuring flask ,add 200 mlglacial acetic acid
(3.3.7 .)and make upto1000 mlwith water .
3.2.6.2 .Solution B - '
Dissolve 290gBaCl^.21^0 A.R. in1000mlofwater .
3.2.7 .Glacial acetic acid 99-100% (lower concentrations are
unsuitable ).
3.2.8 .Ammonium tartrate solution :mix 12.4 gtartaric acid and 12.4 ml
ofammonia solution A.R. (d=0.910 )and make upto1000 mlwith
water (oruse theequivalent amount ofammonium tartrate A.R. ).
3.2.9 .Dilute ammonia solution :40mlammonia solution A.R. (d=0.910 )
diluted to 1000 mlwith water .
3.2.10 .Standard acetate buffer :dissolve 40gsolid sodium hydroxide A.R
in500 mlvratex ina1000 mlvolumetric flask and add 120 ml
glacial acetic acid (3.2.7 .). Mix thoroughly ,cool and make
uptothe .mark with water .

-12
3.2.11 .Pyrrolidinedithiocarbamate solution (shortened to
"Carbate solution ") :dissolve 103 mgsodium Pyrrolidin -
edithiocarbamate (C,H KNa S,2H0)inabout 500 mlwater ,
add 10mln-amyl alcohol A.R. and0.5 gNaHCO ^A.R. ,andmake upto 1000 mlwith water .
3.2.12 .Copper sulphate solution (forstandardization of3.2.11 .).
Stock solution
Mix1,249 gcopper sulphate A.R. (CuSO ,.SH^O )with 50ml0.5 Msulphuric acid and make upto 1000 mlwith water .
Standard solution
Mix 50mlstock solution with 10ml0.5 MH„S0 ,and make up
to 1000 mlwith water .
3.2.13 .Sodium chloride A.R.
3.2.14 .Gas-stripping apparatus (see figure 5).
The diameter ofthe sintered disc must bethe same asthe
internal diameter ofthe cylinder .
3.2.15 .Separating funnel ,250 ml.
3.2.16 .Magnetic stirrer with magnet 25-30 mm.
3.2.17 .Gooch crucible ,diameter oftheperforated base ■»
25 mm,Type G4.
3.2.18 .Circular glass-fibre filter papers ,27 mmdiameter with
fibre diameter 0.5-1.5^im.
3.2.19 . .Twofilter flasks with adaptors and rubber collars ,
500 mland 250 mlrespectively .
3.2.20 .Recording potentiometer fitted with abright platinum
indicator electrode and acalomel orsilver /silver
chloride reference electrode with a250 mVrange ,with
automatic burette of20-25ml capacity ,oralternative manual
equipment .

-13 ->
J3.3 Method -
3.3.1 .Concentration and separation ofthesurfactant
Filter the aqueous sample through aqualitative filter paper .
Discard thefirst 100 mlofthefiltrate .
Into thestripping apparatus ,previously rinsed with ethyl
acetate ,place ameasured quantity ofthe sample ,such thatitcontains between 250-800pgnon-ionic surfactant .
Toimprove theseparation add 100 gsodium chloride and 5g
sodium bicarbonate . ' *
Ifthe volume ofthe sample exceeds 500 mladd these salts tothe
stripping apparatus insolid form ,and dissolve bypassing
nitrogen orairthrough info ffoe .ajDf>aca.tu.s . '
* Ifasmaller-sized sample isused ,dissolve thesalts in400 ml
water and then add tothestripping apparatus .
Add water tobring thelevel tothe upper stopcock .
Cautiously add 100 ml*ethyl acetate ontop .ofthewater .
Fill thewash-bottle inthegas-line (nitrogen orair)two-
vthirds full with ethyl acetate .
Pass agas stream of30-601/hthrough the apparatus ;the
inclusion ofarotameter isrecommended . The rate ofaeration
must beincreased gradually atthebeginning . The gasrate
must besoadjusted that thephases remain noticeably separate
tominimise the mixing ofthephases and thesolution ofthe ;
ethyl acetate inthewater . Stop the gas flow after 5minutes .
Ifthere isareduction ofmore than 20% inthe voluma ofthe
organic phase through solution inwater ,thesublation must
berepeated paying special attention totherate ofgas flow .
Run off the organic phase into aseparating funnel . Return any
water inthe ^separating funnel from the aqueous phase -it
should only beafew ml-tothestripping apparatus .Filter
theethyl acetate phase through adryqualitative filter paper
into a250 mlbeaker .

n14^
Put afurther 100 mlethyl acetate into thestripping
apparatus and again pass nitrogen orair through for
5minutes . Draw off theorganic phase into theseparating
funnel used for thefirst separation ,reject the aqueous
phase and run theorganic phase through the same filter
asthefirst ethyl acetate portion . Rinse both the
separating funnel and thefilter with about 20mlethyl
acetate .
Evaporate theethyl acetate extract todryness oilawater -
bath (fume cupboard ). Direct agentle stream ofair
over the surface ofthe solution toaccelerate the
evaporation .
Precipitation andfiltration
Dissolve the dry residue from 3.3.1 .in5mlmethanol ,add
40mlwater and 0.5 mldiluted HC1 (3.2.3 .)andstir the
mixture with amagnetic stirrer .
Tothis solution add 30mlofprecipitating agent (3.2.6 .)
from ameasuring cylinder ..Theprecipitate forms after
repeated stirring .After stirring for 10min.leave the
mixture tostand for atleast '5min .
Filter the mixture through aGooch crucible ,the base of
which iscovered with aglass-fibre filter paper .First
wash thefilter under suction with "about 2mlglacial
acetic acid . Then thoroughly wash the beaker ,magnet ,
and crucible with glacial acetic acid ,ofwhich about
40-50mlisnecessary .Itisnot necessary toquant­
itatively transfer theprecipitate adhering tothesides of
the beaker ,tothefilter ,because thesolution ofthe
precipitate for thetitration isreturned totheprecipitating
beaker ,and the remaining precipitate will then bedissolved .
Solution oftheprecipitate
Dissolve theprecipitate inthefilter crucible bytheaddition ofhot ammonium ^Vartrate solution (3.2.8 .)in
three portions of10mleach . Allow each portion tostand
inthecrucible for some minutes before being sucked through
the filter into the flask .•
Put the contents ofthefilter flask into the beaker used - ■
for theprecipitation . Rinse the sides ofthe beaker with a
further 20mloftartrate solution todissolve the rest ofthe
precipitate .
# • ,+Carefully wash thecrucible ,adaptoy andfilter flask with
150-200 mlwater ,and return therinsing water tothe beaker
used for theprecipitation .
*)faLo *f8ùcC

3.3.A. The tri«ration
Stir-the solution with amagnetic stirrer (3.2.16 .), add
afew drops ofbromocresol purple (3.2.5 .)and add thediluted
ammonia solution (3.2.9 .)until the colour turns violet (the
solution isweakly'acid from theresidue ofacetic acid used
for rinsing ). .
Then add 10mlstandard acetate buffer (3.2 .10),immerse
theelectrodes inthesolution ,andtitrate potentiometrically
with standard "carbate solution "(3.2.11 .), theburette tip
being immersed .inthesolution . ,
"The titration rate should not exceed 2ml/min .
£
The endpoint'is the "intersection ofthe tangents tothe .
two branches ofthepotential curve .Itwill beobserved
occasionally that theinflection inthepotential curve
becomes flattened ;this can beeliminated bycarefully
cleaning theplatinum electrode (bypolishing with emery paper ).
3.3.5 . Blank determinations
Atthe same time run ablank determination through the whole
procedure with 5mlmethanol and 40mlwater ,according tothe
instructions in3.3.2 . The blank titration should bebelow
1ml.otherwise thepurity ofthe reagents (3.2.3 .-3.2.7 .-
3.2.8 .-3.2.9 .-3.2.10 )issuspect ,especially their content
ofheavy metals^and they must bereplaced . The blank must
betaken into account intije calculation ofthe results .
3.3.6 . Control ofthe factor ofthe "carbate solution "
Determine thefactor for the carbate solution daily . Todothis
titrate 10mlofthe copper sulphate solution (3.2.12 .)with
carbate solution after the addition of100 mlwater and 10ml
standard acetate buffer (3.2.10 ).- Ifthe amount used isJ,a"
ml,the factor fis;
a
andall theresults ofthetitrations 'aremultiplied bythis
factor .

••là
Calculation ofresults
Every non-ionic surfactant hasits ownfactor ,depending
onthe length oftheethylene oxide chain .The concentration of
non-ionic surfactant expressed inrelation toastandard ,substance-anonyl phenol with 10ethylene oxide units (NP10)-for which the
conversion factor is0,054 « Using this factor the amount of
surfactant present inthe sample isfound expressed asmg
ofNP10equivalent ,asfollows
(b-c).f.0.054 ■mgnon-ionic surfactant
where b»volume of"carbate solution "used bythe
sample (ml)
c■»volume of"carbate solution "used bythe
blank (ml)
f=factor ofthe carbate solution "
Expression ofresults
Express theresults asmgBiAS /1tothenearest 0.1.

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