I. Soc. Cosmet.Chem.

, 26, 105-117 (February 1975)

The Influenceof pH, Emulsifier,and

Accelerated
AgeinguponPreservative
Requirementsof OIW Emulsions
GENE JACOBS,M.S., S. M. HENRY, Ph.D., and V. F. COTTY, Ph.D.*
PresentedDecember11, 1973,New York City

Synopsis-Twenty-nine individual PRESERVATIVES and 16 combinations of two or

more preservatives were tested in an ANIONIC and a NONIONIC OIL/WATER
LOTION, each of which was formulated at an acid and an alkaline pH. MINIMUM
INHIBITORY CONCENTRATIONS against four representative challenge MICRO-
ORGANISMS were determined for the preservativesand/or combinations. Less than
35% of the preservativesor systemstested in lotions were effective. Anionic emulsions
were somewhateasierto preservethan formulasmade with nonionicemulsifiers.

INTRODUCTION

A recentsurveyby Gucklhorn(1) reviewsdata on a large numberof anti-

bacterialand antifungalcompounds. He has gatheredinformationon these
compounds from a variety of sourcesand providesan invaluablereview of
the pertinentpublishedinformation.There are, however,seriousgaps in
availabledata due to the variationin testmethodsemployedto evaluatethe
preservatives.
It was our purposeto studythosecompounds of mostinterestin a uniform
system(or systems).In addition,we were interestedin developinga proce-
dure for screeningnew preservatives
that gave more significantresultsthan
the traditional serial dilution in broth. Oil-in-water emulsions,known to be
highly susceptibleto microbialdegradation,were chosenas the model sys-
temsfor thisstudy.

* Bristol-MyersProducts,Hillside, N.J. 07207.

105
106 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

Representative
oil-in-wateremulsions
weredevisedto studythe following
aspectsof preservation:
1. Inaetivation
of preservatives
in anionieandnonionicenmlsifier
systems.
2. The effectof pH on preservation
potential.
3. Stabilityof preservatives
uponageingat elevatedtemperatures.
4. Determinationof Minimum Inhibitory Concentration(MIC) of satis-
factorypreservatives.
5. Augmentation
of activityby inclusionof two or morepreservatives.
A surveyof the literatureon cosmeticpreservation
showsthat severalin-
vestigatorshavestudiedtheinaetivation of preservatives
by nonionicemulsi-
fiers and/or other components of a formulation.Most investigators
use an
aqueous systemto evaluatebindingof preservativesby nonionics.Someex-
cellentpapersof thisgenreareby Patel(2), PatelandKostenbauder
(3),
deNavarre(4), Wedderbum(5), andBarrandTice (6).
A numberof papershave been publishedwhich deal with interactions
betweenanioniesandpreservatives. Manyof thesestatethat anionieemulsi-
fiersmayenhance the activityof somepreservatives.
However,Schuster and
Modde (7) demonstrate bindingof somepreservatives by anioniemoieties
in aqueous systems.
Entrekin (8) and Wiekliffeand Entrekin (9) evaluatedthe effectsof pH
on preservatives
in a brothmedium.OptimalpH conditions wereascertained
for 17 preservatives
and 4 preservativemixtures.
Oil-in-wateremulsionshave been usedby a few authorsto evaluatepres-
ervationphenomena.Boehm(10) usedemulsions to evaluatesynergism
be-
tweenpreservatives
againsta varietyof testorganisms. BarrandTiee (6)
studiedthe preservation
of aqueoussolutionscontainingnonionicsurfae-
rants.Thosepreservativesshowingsatisfactory resultswere incorporated
intoemulsions
to comparepreservative
activityin aqueous systemswithemul-
sions.
The effectof the hydrophilie-lipophilie
balance(HLB) of nonionicsur-
factantson preservativesin emulsionswas studiedby Tilbury (11). Ten
preservatives
were incorporatedinto simpleemulsionsto evaluateHLB on
preservation.
Beanet al. (12) studiedthe effectsof the partitioncoefficient
of a preservativein cosmeticemulsions.
The presentstudy differsfrom mostof the aboveinvestigations in that it
is concernedwith the simultaneous evaluationof pH, emulsifiertype, and
acceleratedageingof completeoil-in-wateremulsionspreparedwith the great
variety of preservatives
now available.Further, it is an extensionand up-
datingof severalfacetsof the preservativestudiesconductedby others.Pre-
servativesno longer usablein cosmeticsbecauseof regulatoryaction have
been excluded from evaluation. The authors,instead, have concentratedon
thosecurrently available to us including many new compoundswith little
or no historyof usein oil-in-wateremulsions.
The tablesof effectivelevels
 PRESERVATIVEREQUIREMENTS OF O/W EMULSIONS 107

of the various
compoundstakeintoconsideration
notonlystabilityat high
temperature but suggest
approximate
minimum concentrations
of preserva-
tivesandpreservative
combinations
foroil-in-water
emulsions
of various
types.
EXPERIMENTAL

The testorganisms
were:$treptococcus
)•aecalis
(ATCC4082),Pseudo-
monasaeruginosa
(ATCC 15442),Candidaalbicans(ATCC 10231),and
Aspergillusniger ( ATCC 9642).
The bacteriaweregrownin Tryptica•se
SoyBroth(BBL) for 24 hoursat
37øC;theyeastin MycophilBroth(BBL) for 24 hoursat 37øC;thefungus
on MycophilAgarslantsfor 10-14 daysat 30øC.Fungalspores werehar-
vestedfrom10-dayold slantswith steriledistilledwaterandthissuspension
usedfor challenge.
Testlotionformulasusedin the studyarelistedin TableI.
Dataonpreservatives
andpreservative
mixtures
tested
aregivenin Tables
II-IV. The chemicalname,trade name,and sourceof eachof the preserva-
tives are included.

Table I

Formulas for Anionic and Nonionic Test Lotions

Preparations Acid Alkaline

Anionic Lotions
Carbopol941 0.15 0.15
Cetylalcohol 0.50 0.50
Glycerylmonostearate,
NSE 2.50 2.50
Isopropyl-palmitate-myristate
(60-35) 2.00 2.00
Mineral Oil 55/65 SUS 1.00 1.00
Sodiumlauroylisethionate 0.50 0.50
Triethanolamine,
98% 0.05 0.40
Glycerin,anhydrous 4.80 4.80
Water deionized 88.50 88.15
•00.00 100.00
pH 5.1-5.2 pH 7.9-8.1

Nonionic Lotions
Mineral Oil 55/65 SUS 20.00 20.00
Cetylalcohol 5.00 5.00
Span60 2.50 2.50
Tween 60 7.50 7.50
Sodiumphosphate,dibasic,anhydrous 0.01 ...
Sodiumphosphate, tribasic,anhydrous ... 0.12
Water, deionized 64.99 64.88
i00.00 100.00
pH 5.4-5.7 pH 7.9-8.3
108 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

Table II

Minimum Inhibitory Concentration(%) for Preservatives

Effectivein One or More Lotions
Preservative Anionic Lotion a Nonionic Lotion a

Trade Name Chemical Name Acid Alkaline Acid Alkaline

Bronopol
(Goldschmidt) 2-Bromo-2-nitropropane-1-3-diol 0.05 > 0.1D 0.1 > 0.1•)
Dehydroacetic 3-Acetyl-6-methyl-2H-pyran-2,
acid (Gane's) 4 (3H)-dione 0.1 NT '>0.2 NT
Dioxin -•
Giv-Gard DXN 6-Acetoxy-2,4-dimethyl-m-
(Givaudan) dioxane 0.1 0.1 0.15 0.2
DMDMH (Glyco) Dimethylol dimethylhydantoin 0.1 0.1 >0.1 0.1
Dowicil 200 Cis isomerof 1-(3-chloroallyl)-
(Dow) 3,5,7-triaza-l-azonia-adaman-
tane chloride 0.075 D 0.1 I} 0.1 0.1
Formaldehyde
(Fisher) Formaldehydesolution,40% 0.05 0.05 0.075 0.675
Hibitane Bis (p-chlorophenyldiguanido)
(Imperial) hexane >0.1 >0.1 0.1 >0.1
MDMH (Glyco) Monomethyloldimethyl
hydantoin 0.2 0.2 >0.5 >0.5
Nipastat (Nipa) Nipa ester82121-mixture of
methyl, ethyl, propyl, butyl,
benzyl parabens 0.225 >0.3 >0.3 >0.3
Noxyfiex N-Methyl-N'-hydroxymethyl
(Geistlich} thiourea 0.25 0.2 0.5 0.2
Ottasept (Ottawa) Parachlorometaxylenol >0.5 0.38 >0.5 >0.5
Phenonip(Nipa) Mixture of phenoxetoland
parabens 0.5* >0.5 >0.5 >0.5
Phenoxetol
(Nipa) Ethylene glycolmonophenyl
ether 1.0 1.0 1.0 >1.0
Phenylc'hyl
alcohol(IFF) 2-Phenylethanol 1.0 1.0 >1.0 >1.0
PolycideA Tris (2-hydroxyethyl)triazonium
(Zwicker) methyl carboxyphenolate 0.15 0.1 0.15 0.1
Polynoxylin
(Geistlieh) Polynoxymethyleneurea >0.3 >0.3 >0.3 0.3
Sorbic acid
(Pfizer) 2,4-Hexadienoicacid 0.1 NT 0.15 NT
Omadine, Sodium
(Old) Sodiumpyridinethione C.I' >0. l 0.1' 0.1 •

Symbolsused: D = Discoloration;NT: Not Tested; * = ApproximateMIC; >n: In-

effective at the maximum concentration indicated.
 PRESERVATIVE REQUIREMENTS OF O/W EMULSIONS 109

Table III

Minimum Inhibitory Concentrations(%) for PreservativeMixtures

Effective in One or More Lotions

Preservative Mixture Anionic Lotion a Nonionic Lotion a

Trade Name Chemical Name Acid Alkaline Acid Alkaline

Bronopo1-4- 0.1 •' 0.P >0.1 >0.1

Omadlne, Sodium 0.05 0.05 >0.05 >0.05
LMDMII -4- 0.1' 0.1 * 0.1' 0.1 •
Omadine, Sodium 0.05 0.05 0.05 0.05
MDMH q- 0.2* 0.2* 0.2* 0.2*
Omadine, Sodium 0.05 0.05 0.05 0.05
Bronopolq- 0.018 0.013 • 0.005 >0.1 •>
Methylparaben Methyl p-hydroxy-
(Mallinckrodt) + benzoic acid 0.038 0.025 0.1 >0.2
Propylparaben Propyl p-hydroxy-
(Mallinckrodt) benzoic acid 0.009 0.00e 0.025 >0.05
Formaldehydeq- 0.013 0.018 0.038 0.038
Methylparabenq- 0.05 0.075 0.15 0.15
Propylparaben 0.013 0.018 0.038 0.038
Germall q- 0.38 0.5 0.5 0.5
Methylparabenq- 0.15 0.2 0.2 0.2
Propylparaben 0.038 0.05 0.05 0.05
G•rmall q- 0.5* 0.5* >0.5 >0.5
Methylparabenq- 0.1 0.1 >0.1 >0.1
Butylparaben 0.02 0.02 >0.02 >0.02
Methylparabenq- 0.15 >0.2 >0.2 >0.2
Propylparaben 0.038 >0.05 >0.05 >0.05
Hibitane q- >0.1 >0.1 0.1 >0.1
EDTA (Ciba-Geigy) Ethylene diamine- > 0.05 > 0.05 0.05 > 0.05
tetraacetic acid
Vancide89RE n-Trichloromethylthio-
(Vanderbilt)q- 4-cyclohexene-l,2-
dicarboximide >0.2s 0.075 >0.2v >0.2v
Phenoxetol > 1.0 0.375 > 1.0 > 1.0
Preservative68 Halogenatedaliphatic
(Dutton
& Reinisch)
q- amide 0.13 0.19 0.25 0.19
Formaldehyde 0.013 0.019 0.025 0.019
Symbolsused:* = Approximate
MIC; D = Discoloration;
S= Separation
of emulsion;
V = Viscosity
changes;
>n: ineffective
at themaximum
concentration
indicated.

Test Procedures

Preparationof Lotions
Testlotionswithoutpreservatives
werepreparedand immediatelyrefrig-
eratedto preventspoilage.
Preservatives
weredissolved
in wateror ethanol,
11o JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

Table IV

Preservatives
and PreservativeMixturesUnsatisfactoryin All Lotions Tested
Preservativeor PreservativeMixture Maximum
Level

TradeName Chemical
Name Tested
(%)

Armeen Z (Armour) n-Coco-B-aminobutyricacid 0.1

Bradosol(Ciba-Geigy) Dodecyl-dimethyl (2-phenoxyethyl)
ammonium bromide 0.1
Dowicide 1 (Dow) Orthophenylphenol 0.2
DragocidForte (Dragoco) Isopropylsorbate 1.0
Germall 115 (Sutton) Imidazolidinylurea 0.5
Omadine, Zinc (Olin) Zinc pyridinethione 0.1
Parablend(Mallinckrodt) Mixture of methyl, ethyl, propyl,
butyl parabens 0.2
Preservative68 (Dutton & Reinisch) Halogenatedaliphaticamide 0.5
Propyleneglycol (Union Carbide) 1,2-Propanediol 10.0
Tego 103S (Goldschmidt) 15% Dodecyldi(aminoethyl)glycine 0.5
Vancide 89BE (Vanderbilt) n-Trichloromethylthio-4-cyclohexene-
1,2,-dicarboximide 0.2
MDMH + O.5
Methylparabenq- 0.25
PropylI)araben 0.05
Methylparaben q- 0.1
Butylparaben
(Mallinckrodt) Butylp-hydroxybenzoic acid 0.02
Irgasan-Triclosan
(Ciba-Geigy)q- 2,4,4'-Trich]oro-2'-hydroxy-diphenyl
ether 0.1
EDTA 0.05
Vancide 89RE q- 0.2
EDTA 0.05
Vancide 89BE q- 0.2
Formaldehyde 0.05

accordingto their solubilitiesand addedto 95 ml of lotionin a 4-ouncescrew-

cappedjar. The finalvolumeof preserved
lotionwasbroughtto 100ml by
the additionof sterilewater, where necessary. After additionof preserva-
tive, the lotionwasallowedto equilibrateat roomtemperature for 3-7 days
and shakenoccasionally. Four 10-ml aliquotswere placedin 9,0x 150 mm
steriletest tubes.The remaininglotionwas storedat 5ϿCfor one month,
afterwhichtimeit wasre-examined for preservative
adequacy.
Challengeof Lotions
Ten-milliliteraliquotsof each test lotion were challengedwith the fol-
lowingvolumesof purecultures:
Streptococcus faecalis-O.09,ml of 24-hourbroth culture (9,.7x 107 orga-
nisms/10ml product)
 PRESERVATIVE REQUIREMENTS OF O/W EMULSIONS 111

Pseudomonas aeruginosa-O.02ml of 24-hour broth culture (3.7 x 107 or-

ganisms/10ml product)
Candidaalbicans-O.05ml of 24-hourbroth culture (3.5 x 10• organisms/
10 ml product)
Aspergillusniger-0.1 ml of a freshlypreparedsporesuspension (4.5 x 105
organisms/10 ml product)
P. aeruginosa,C. albicans,and A. niger were recoveredin large numbers
in all unpreservedtestlotionsfor at least14 daysafter challenge.S. faecalis
survivedin high numbersin all lotionsexceptthe acid anionic,where no
organisms were detectedafter a 3-daycontactperiod.

Subcultures
of ChallengedLotions
At the followingtimeintervalsafter challenge,eachsamplewasthorough-
ly mixedand a sterileinoculatingloop of ca. 0.05-mlcapacitywas usedto
streakthe lotionontoa Letheenagar (Difco) plate--0hour,3 days,and 7
days.
Bacterialandyeastplateswereincubatedat 37øCfor a maximum
of 3 days;
fungalplateswere incubatedat 30øCfor 3-7 daysbeforediscarding.

Storageof ChallengedLotions
Lotionsinoculatedwith challengeorganismswere stored at 24øC during
the subcultureperiod.After the finalsubculture,they were discarded.
Stagesin the Evaluationof Preservative
Adequacy
FreshlyPreparedUnagedLotions-Threedecimaldilutionsof eachpre-
servativewere incorporated into eachlotion to determinethe approximate
level of preservative
required.The highestconcentrationtestedwas usually
that recommendedby the supplieror noted in the literature;however,in
some cases,the maximum level tested was twice the recommendedlevel. Kill
or markedreductionof 3 or more challengeorganismswithin 3 days of in-
oculationwas consideredmandatoryfor additionalstudy.Recoveryof high
levelsof organismsat any preservativelevel precludedfurther examination
of that level.
Aged Lotions-Samplesstored at 52øC for one month were subjectedto
challengeby the 4 test organismsif they had been proven satisfactoryin
preservativeactivityby the previoustest.Kill of all challengeorganisms
with-
in 3 days indicated potentially adequatepreservationafter storageat ele-
vated temperature.The satisfactory levelsdeterminedby the precedingtwo
studiesbecamethe startingpoint for MIC studies.Theselevelswere desig-
nated "effective levels."
MIC Studies-Preservativeswere incorporatedinto freshlypreparedtest
lotionsin 25% decrementsfrom the "effectivelevel" determinedin aged lo-
11'2 JOURNAL OF THE SOCIETY OF COSMETIC CItEMISTS

Table V. Effects of Combining Preservativeson Effective Concentrationsand Microbial

Spectrum (Individual PreservativesPrecedePreservativeMixtures)
Anionic a Nonionic a

Acid Alkaline Acid Alkaline

Preservative -Result % Result % Result % Result %

Bronopol S 0.05 Ue 0.1 S 0.1 Ue 0.1
Sodium Oreadinc S 0.1 U s.a 0.1 S 0.1 S 0.1
Bronopolq- S 0.1 S 0.1 U e,a 0.1 Ue,a 0.1
Sodium Oreadinc 0.05 0.05 0.05 0.05

Methylparabenq- S 0.15 Us 0.2 Ua 0.2 Us,p,a 0.2

Propylparaben 0.04 0.05 0.05 0.05
Bronopol q- S 0.018 S 0.013 S 0.05 Ua 0.1
Methylparabenq- 0.038 0.025 0.1 0.2
Propylparaben 0.009 0.006 0.0•5 0.05
Formaldehyde S 0.05 S 0.05 S 0.075 $ 0.075
Formaldehydeq- S 0.013 S 0.018 S 0.038 S 0.03S
Methylparaben
q- 0.05 0.075 0.15 I 0.15
Propylparaben 0.013 0.018 0.038 0.038
Gemsall U e,a 0.5 U c,a 0.5 U e,a 0.5 U e,a 0.5
Germall q- S 0.38 S 0.5 S 0.5 S 0.5
Methylparaben q- 0.15 0.2 0.2 0.05
Propylparaben 0.038 0.05 0.05 Us,l•,c,
a 0.1
Methylparabenq- Us,e,a 0.1 Us,P,e,
a 0.1 Us,v,e,a 0.1 0.02
Butylparaben 0.02 (I.02 0.02 Ue 0.5
Germall q- S 0.5 S 0.5 Ue 0.5 0.1
Methylparabenq- 0.1 0.1 0.1 0.02
Butylparaben 0.02 0.02 0.02 0.2
MDMH NT NT Uc 0.5 Uc 0.5
MDMH q- NT NT Uc 0.,5 Ue 0.5
Methylparabenq- 0.25 0.25
Propylparaben 0.05 0.05
Vancide 89RE U* 0.2 Up,e,a 0.2 Up,a 0.9. UP,a 0.2
Phenoxetol S 1.0 S 1.0 S 1.0 U* 1.0
Vancide 89RE q- U* 0.2 S 0.075 U* 0.2 U* 0.2
Phenoxetol 1.0 0.375 1.0 1,0

Preservative 68 Us 0.5 Us 0.5 Us,P 0.5 Us,v 0.5

Preservative68 q- S 0.13 S 0.19 S 0.25 S 0.19
Formaldehyde 0.013 0.019 0.025 0.019

Symbolsused: * = Organolepticproblems;S = Satisfactory;U = Unsatisfactory;NT:

Not Tested. Subscriptfigure denotessin'rival of: s :- S. [aecalis;p: P. aeruginosa;c:
C. albicans;a = A. niger.
 PRESERVATIVE REQUIREMENTS OF O/W EMULSIONS 113

tions.The level of preservativewas reduceduntil the minimuminhibitory

concentrationfor the most resistantorganismwas attained.The level that
killedall organisms
•vithin7 dayswasconsideredto be the MIC.
RESULTS

Table II summarizes
M[C dataobtainedon prescrvatives
effectivein one
or more lotions.MIC data for preservativemixtureseffectivein one or more
lotionsappear in Table III. Those preservativesand preservativemixtures
foundto be unsatisfactoryin all lotionstestedare listedin Table IV.
Organolepticobservations are included and noted as superscripts where
applicable.•Ihey are explainedat the baseo[ the tables.In somecases(no-
tably Vancide89RE combinations), deteriorationof aged lotionsprevented
furthertestingof lotionswhichwereinitiallysatisfactory.
In preparingpreservativemixtures,the basicapproachtaken was to com-
bine t;vo or more preservatives so as to eliminategapsin m'•crobko_ogical
spectrum,thereby accomplishing a complementary or synergisticmode of
activityfor the mixture.Table V presentsdata which illustratethis approach
by comparingpreservative activityof eachcompoundwhentestedaloneand
when in combination.Only thosecombinations
which showsomekind of in-
teractionare presented.Organismsunaffectedby the preservativeand/or
the mixtureare indicatedby superscripts.
Resultsof mixturesrecommended
by the manufacturers,suchas Germallplus parabens,are also reportedin
Table V.
Table VI summarizesthe resultsof 174 preservationstudiesconducted
on 4 lotion types.A total of 57 lotions(33%) were adequatelypreserved.
Of these 57 lotions, 33 were anionic emulsionsand 24 were nonionic. These
resultsindicate a greater easein preservinganionie emulsions.Acidic an-
ionic lotionswere satisfactorilypreservedin 18/44 eases(41%); alkaline
nonioniclotionswere the mostdifficultto preserveof the 4 lotion types-
10/44 satisfactory(23%).
Table VI

PreservationAccording to Lotion Type

Number Number Per cent
Lotion Type Tested Satisfactory Satisfactory
__

Acid anionic 44 18 41
Alkaline anionic 42 15 36
Total anionics 86 33 38

Acid nonionic 45 14 31
Alkaline nonionic 43 10 23
Total nonionics 88 24 27

All lotions 174 57 33

114 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

DISCUSSION

When a preservativeor preservativemixturefailed in one or morelotions

(Tables II-IV), the failure mostoften occurredin the unagedlotion. How-
ever, in about 30% of the tests, elevated temperatureand/or storagefor
one month appearedto be responsiblefor lossof preservativeactivity.
Severalcombinationsof preservativesare effectivein one or more lotions
by virtue of complementary activity againstthe test organisms. The combi-
nationof Germallwith parabenstypifiesthis type of activity.There is also
apparentsynergism asin the caseof Bronopolplusparabens.Somecombina-
tionsprovidepartial complementation. EDTA kills P. aeruginosain 3 of the
4 Irgasan-EDTAmixturestested,but failureto kill all challengespecies makes
thiscombination ineffectivefor any of the lotions.Finally, thereis at leastone
exampleof apparentantagonism (BronopolplussodiumOmadinein the acid
nonioniclotion). It is obviousthat combinations
of compoundsmay provide
adequatepreservation of formulasin which singlepreservatives
are inef-
fective.However,one cannoteasilypredictwhethera theoreticallyaccepta-
ble preservative
mixturewill be effectivein a formulation;additiveeffects,
Table VII

PreservativesRecommendedfor VariousLotion Types

MIC Level (%)
Anionic Lotion a Nonionic Lotion a

Preservative Acid Alkaline Acid Alkaline

Bronopol 0.05 NR 0.1 NR

Dehydroaceticacid 0.1 NT NR NT
Dioxin (Giv-Gard DXN) 0.1 0.1 0.15 0.2
DMDMH 0.1 0.1 NR 0.1
Dowicil 200 0.075 D 0.1 l) 0.1 0.1
Formaldehyde 0.05 0.05 0.075 0.075
Hibitane NR NR 0.1 NR
MDMH 0.2 0.2 NR NR
Nipastat 0.225 NR NR NR
Noxyfiex 0.25 0.2 0.5 0.2
Ott•asept NR 0.38 NR NR
Phenonip 0.5* NR NR NR
Phenoxetol 1.0 1.0 1.0 NR
Phenylethylalcohol 1.0 1.0 NR NR
PolycideA 0.15 0.1 0.15 0.1
Polynoxylin NR NR NR 0.3
Sorbic acid 0.1 NT 0.15 NT
Oreadinc,sodium 0.1' NR 0.1' 0.1'

aSymbolsused: NR = Not recommended;NT = Not tested; D: May discolor;* = Ap-

proximate MIC.
 PRESERVATIVE REQUIREMENTS OF O/W EMULSIONS 115

synergism,or antagonismmay occur.Experimentalevaluationis alwaysnec-

essary.
Our results generally agree with the available literature, i.e., adequate
preservationis more easily accomplished in anionic systemsthan in non-
ionic systems.Further, acidic conditionsallow for the use of a greaternum-
ber of preservatives
than alkalineconditions.
Tables VII and VIII showpreservatives and preservativecombinations
recommendedfor specificlotion types. The concentrations of preservative
givenrepresentthe MIC obtainedby the variousmethodsemployedin this
study.Note, that in practice,concentrations higherthan the MIC wottld be
employedto compensate for possiblelossof preservativedueto heat,light,or
interactionwith otherproductcomponents.
Table VIII

Preservative
Combinations
Recommended
for VariousLotionTypes
MIC Level (%)
Preservative Combination Anionic Lotion a Nonionic Lotion a

Acid Alkaline Acid Alkaline

Bronopolq- 0.1' 0.1' NR NR

Omacline,sodium 0.05 0.05
DMDMH q- 0.1' 0.1' 0.1' 0.1'
Oreadinc, sodium 0.05 0.05 0.05 0.05
MDMH q- 0.2* 0.2* 0.2* 0.2*
Oreadinc, sodium 0.05 0.05 0.05 0.05
Bronopolq- 0.01•8 0.013TM 0.005 NR
Methylparabenq- 0.038 0.025 0.1
Propylparaben 0.009 0.006 0.025
Formaldehydeq- 0.013 0.018 0.038 0.038
Methylparabenq- 0.05 0.075 0.15 0.15
Propylparaben 0.013 0.018 0.038 0.038
Germall q- 0.38 0.5 0.5 0.5
Methylparabenq- O.15 0.2 0.2 0.2
Propylparaben 0.038 0.05 0.05 0.05
Germall q- 0.5* 0.5* NR NR
Methylparabenq- 0.1 0.1
Butylparaben 0.02 0.02
• lethy]parabenq- 0.15 NR NR NR
Propylparaben 0.038
Hibitane q- NR NR 0.1 NR
EDTA 0.05
Vancide 89RE q- NR 0.075 NR NR
Phenoxetol 0.375
Preservative68 q- 0.13 0.19 0.25 0.19
Formaldehyde 0.013 0.019 0.025 0.019

Symbolsused:NR = Not recommended;

* --=ApproximateMIC; D = May discolor.
116 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

The authorswouldbe guiltyof extremenegligence

weretheynotto point
out that the data givenhere cannotbe extrapolatedto all oil-in-wateremul-
sions.Thesedata were obtainedusingmodelsystems with relativelylow
levelsof emulsifiersAny significantincreasein concentration
of emulsifier
may requirean increasein concentration
of preservativeor may even contra-
indicatean otherwisesatisfactory preservativeor preservative
system.Per-
haps,to a lesserdegree,particulartypesof anionicor nonionicemulsifiers
will alsodeterminewhetheror not a preservative
is effective.
Hence,the "effective"levelsof preservatives
givenin this report shouldbe
consideredonly as a guide to be interpretedaccordingto the particular
formulationin question.Followinga selectionbasedon typesand concentra-
tionsof emulsifiers, pH, and otherpertinentcharacteristics
of the formula,the
suitability •of the preservativemust be demonstratedby a satisfactoryre-
sponsein a challengeor preservationtest (13-15) if the formulatoris to be
assuredof a microbiologically acceptableproduct.
Finally, the authorswish to emphasizethat rather strict standardsof ac-
ceptancehave prevailed in this study, viz., lessthan 7 days survivalat the
MIC and lessthan 3 daysat higher concentrations. Althoughlesssevcrere-
quirements,e.g., a longer survival time or microbiostasis of one or more
species,would resultin a greaternumber of acceptablecompoundsand re-
duced levels of preservativesin a formula, the strengthof the protection
aga-'nstmicrobialspoilagewouldbe greatlyreduced.
ACKNOWLEDGlX•ENTS

The authorsgratefully acknowledgethe suggestions of Dr. Arnold D.

Marcus and the cooperationof Messrs.Ara Nersesian,Richard Alonso,Fred
Hubner, and TarasDurbak in developingand formulatingthe lotionsusedin
thisinvestigation.
(ReceivedJanuary15, 1974)
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