j. Cosmet,
sci., 57, 127-137 (March/April 2006)
Analysisof consumercosmeticproductsfor
phthalateesters
JEAN C. HUBINGER and DONALD C. HAVERY, U,S. Food
and Drug Administration,
5100 Paint BranchParkway,College
Park,
MD 20740.
Accepted
for publication
November
17, 2005.
Synopsis
A rapid and sensitivereverse-phase
HPLC method with UV detectionwasdevelopedfor the quantitation
of dimethyl phthalate (DMP), diethyl phthalate(DEP), butyl benzyl phthalate (BBP), dibutyl phthalate
(DBP), anddi(2-ethylhexyl)phthalate(DEHP) in cosmeticpreparations.
Averagerecoveries
of the phthalate
esterswerebetter than 90%. In a surveyof 48 consumer
cosmeticproducts,includinghair careproducts,
deodorants,
lotionsand creams,nail products,fragrances,
and bodywashes,most productswere found to
containat leastonephthalateester.DEP wasdetectedmostfrequentlyat concentrations
up to 38,663 ppm.
DBP wasfound in fewerproducts,but at levelsup to 59,815 ppm. Basedon the availableexposureand
toxicity data, the FDA hasconcludedthat there is insufficientdata to concludethat a human health hazard
existsfrom exposureto phthalateestersfrom cosmeticproducts.
INTRODUCTION
Phthalateestersarepresentin many consumerproducts,primarily to impart flexibility
to rigid polymerssuchaspolyvinylchloride.Theyareusedin the productionof products
suchasfoodwrappings,medicaldevices(e.g.,bloodbags),children'stoys,woodfinishes,
paints, upholstery,and plastic products,and are subject to a variety of regulatory
requirements.
As a resultof their extensiveuse,phthalateestershave beenfound in
the environment(1-6), foods(7-9), food supplements(10), medicalproducts(11,12),
medicaldevices(13), plasticmaterials(14), and cosmetics(15). In cosmeticproducts,
phthalateestersare usedas solventsfor fragrances,as suspension
agentsfor solidsin
aerosols,
aslubricantsfor aerosolvalves,and asantifoamingagents,skin emollients,and
plasticizersin nail polishesand fingernail elongators.
Investigationsinto the levelsof phthalateestermetabolitesin humanurine haveshown
that exposure
to DEP, BBP, DBP, and DEHP from all sources
is highly variablefrom
personto personand betweendifferent demographicgroups(16-19). In one study,
womenof child-bearingage(20-40 years)werefoundto havesignificantlyhigher levels
of monobutylphthalate,the metaboliteof DBP, in their urine than other age/gender
groups(16). Severalphthalatemetaboliteshavealsobeenfoundin humanbreastmilk
127
128
JOURNAL OF COSMETIC SCIENCE
(20). Estimatesof daily human exposureto DEP, basedon estermetaboliteconcentrations in urine, rangefrom undetectableto 320 mg/kg/day(21).
Phthalateesters,are of potential concernbecauseof reproductiveand other toxic effects
reportedfor phthalate estersin animal models.Tests with rats have shownthat some
phthalateesterscandamagethe male reproductive
systemof offspringand causeother
developmentalabnormalities(22-36).
The relevanceof human exposureto phthalateestersis difficult to assess
becausethe
effectsobserved
in animalsresultedfrom exposure
to relativelyhigh doses(on the order
of 100 to 1000 mg/kg/day),andmoreimportantly,ratsmaymetabolizephthalateesters
differentlythanhumans(37,38). While onestudyusinga cometassaysuggested
that the
urinarymetaboliteof DEP at levelscurrentlyfound in the environmentmight cause
human spermDNA damage(39), anotherstudyshowedprimateswere lesssensitiveto
phthalateexposurethan rodents(40). Further, becauseof their ubiquitoususe, it is
difficult to pinpoint any specificsourceof phthalateesterusesas being potentially
responsible
for any observedeffects.
In July 2002, the EnvironmentalWorking Group(EWG), a coalitionof environmental
andpublichealthorganizations,
reportedon the analysisof 72 cosmeticproductsfor the
phthalate estersDMP, DEP, BBP, DBP, and DEHP (15). According to the report,
phthalateesterswere found in 52 of the 72 productstested,at levelsrangingfrom less
than 50 partsper million to nearlythreepercent.Also, accordingto the report,noneof
the 52 productslistedthe phthalateestersasingredientson the productlabels.Basedon
the safetyconcernsraisedby the EWG and other publisheddata, the FDA initiated a
projectto determineconsumer
exposure
to phthalateestersfrom cosmetic
products(41).
The presentstudy was undertakento developand validatean analyticalmethod for
phthalateestersin cosmeticproducts,to verify the levelsreportedin cosmetics
by the
EWG, and to collectadditionaldataon phthalateesterlevelsin othertypesof cosmetic
products.
A largevarietyof analyticalmethodshavebeenpublishedfor the analysisof consumer
products,biologicalmaterials,and environmentalsamplesfor phthalateesters.Methods
basedon gaschromatography
with eitherflameionizationor electroncapturedetection
havebeendescribed
for foodsimulants(42), water(43), plasma(44) andedibleoils(45).
High-performanceliquid chromatography
(HPLC) with UV detectionhasbeenapplied
to water(46), IV drug solutions(47), bloodplasma(48), andwater(49). More recently
reportedmethodshaveutilized massspectrometryfor the detectionof phthalateesters.
Methodsapplying gas chromatographycoupledto a massspectrometerhave beendescribedfor the analysisof water (50-53), saliva(54), and plastic and PVC materials
(55,56). HPLC andmassspectrometry
havebeenusedfor the analysisof urine(57-59),
human milk (20), and IV drug solutions(60). Very little hasbeenpublishedon analyticalmethodsfor the determinationof phthalateestersin cosmeticproducts.Two older
methods,basedon simplegaschromatography,
havebeendescribedfor cosmeticproducts(61,62), but little datahasbeenprovided.The presentstudydescribes
a methodfor
the determinationof phthalateestersin a variety of differentcosmeticproducts.The
methodutilized a Celite columnextractionmethodoriginallydevelopedin our laboratory for the analysis
of phenol,resorcinol,
salicylicacid,ando•-hydroxy
acidsin cosmetic
productsand salonpreparations(63). The methodwasvalidatedfor the determination
of phthalateestersin severaldifferenttypesof cosmeticproducts.
PHTHALATE
ESTERS IN COSMETIC
PRODUCTS
129
EXPERIMENTAL
REAGENTS
AND
MATERIALS
The followingreagentsand materialswere used:Hexanewaspurchasedfrom Burdick &
Jackson(Muskegon,Michigan). Acetonitrileand methanolwere purchasedfrom T. J.
Baker (Phillipsburg, New Jersey).2-Propanol was purchasedfrom Fisher Scientific
(Fairlawn,New Jersey).All solventswere HPLC grade. PhthalateestersDMP (99%),
DEP (99.5%), and BBP (98%) were purchasedfrom Sigma Aldrich (Milwaukee, Wisconsin).DBP (-->98%) and DEHP (99.5%) were purchasedfrom Sigma Aldrich (St.
Louis,Missouri).De-ionizedwaterwaspreparedwith a Milli-Q purificationsystemfrom
Millipore (Billerica, Massachusetts).
Celite 545 was purchasedfrom Fisher Scientific
(Fairlawn,New Jersey).The extractiontubesandfilter diskswereobtainedfrom Supelco
(Bellefonte,Pennsylvania).
PHTHALATE
ESTER
CALIBRATION
STANDARDS
A primary standardsolutionof a mixture of the five phthalateesters(- mg/ml each)was
preparedby addingapproximately100 mg of eachto a 100-ml ambervolumetricflask
and diluting to the mark with hexane.Becauseof the wide range of possibleconcentrationsin cosmeticproducts,threesetsof workingstandards
wereprepared.One setwas
preparedat approximately0.001, 0.003, 0.006, and 0.01 mg/ml by appropriateserial
dilution of the stocksolution.Similarly, a secondset was preparedat approximately
0.01, 0.03, 0.06, and 0.1 mg/ml, and a third set for BBP, DBP, and DEHP only was
preparedat 0.10, 0.30, 0.60, and 1.00 mg/ml. HPLC peakareaswere determinedbased
on duplicateinjectionsof 20 pl, and a calibrationcurvewasobtainedby plotting peak
area versus standard
SAMPLE
concentration.
EXTRACTION
To avoidcontaminationby environmentalsources
of phthalateesters,all glassware
was
thoroughly cleanedand rinsed with water and ethanol before use, and phthalatecontainingplasticswere avoided.Approximately 1 g of each cosmeticsamplewas
weighedinto a 40-ml beaker,mixed thoroughlywith about 3 g of Celite, and then
transferredto a 15-ml extractiontube containinga filter disk. The sample/Celitemixture wascoveredwith a secondfilter disk and compacted
firmly with a stirringrod. The
preparedcolumn was eluted with sufficienthexaneto obtain 10 ml of extract in a
volumetricflask.The extractionflask wasmixed well prior to HPLC analysis.
HPLC
ANALYSES
HPLC analyseswere carriedout on an Agilent 1100 seriesHPLC, equippedwith a
quaternarypumping system,an in-line vacuumdegasser,
a variablewavelengthdiode
arrayUV-visible absorbance
detector,a 20-121injectionloop, and a personalcomputer
with
HP
Chemstation
software.
Chromatographicseparationwasachievedusinga Whatman Partisil ODS-3 5-12mguard
column (7.5 mm by 4.6 mm ID) and a Whatman Partisil ODS-3 5-12manalytical
130
JOURNAL OF COSMETIC SCIENCE
column(250 mm by 4.6 mm ID), both obtainedfrom Altech Chromatography
(Deerfield, Illinois). The analyticalcolumnwashousedin a thermostattedcolumncompartment at room temperature.Chromatographicseparationwas achievedusing a solvent
programstartinginitially at 50% water, 34% acetonitrile,13% 2-propanol,and 3%
methanolthat waschangedlinearly over 35 minutesto 15% water, 55% acetonitrile,
25 % 2-propanol,and 5% methanol,and held at the final compositionfor an additional
ten minutes. The mobile phaseflow rate was 1.0 ml/min. The mobile phasewas
graduallyreturnedto the initial mobilephasecomposition
overa periodof ten minutes.
Phthalate esters were detected at 230 nm.
QUANTITATION
Four-point calibrationcurveswere preparedfor eachphthalateester. Phthalateswere
identified in sampleextractsby comparingHPLC retentiontimes with standards,and
quantitatedusingthe standardcalibrationcurvefor eachphthalateester.Sampleextracts
werediluted asnecessary
to confirmthat the concentrations
werein the linearrangeof
the calibrationcurve.Calculatedphthalateesterconcentrations
of 10 ppm or lesswere
recorded as not detected.
RECOVERY
The recoveryof phthalateestersfrom cosmeticproductswasdeterminedby fortifying
productswith 100 and 1,000 ppm of each ester followed by extractionand HPLC
analysisas describedabove.
RESULTS
AND
DISCUSSION
In this study,a rapidmethodfor the determinationof fivephthalateesterswasdeveloped
and validated.For most cosmeticsampleextracts,eachphthalateesterwas completely
separated
from othercomponents
by HPLC and couldbe quantitatedunambiguously.
To furtherconfirmthat observed
peaksweredue to phthalateestersand not impurities,
UV spectraof chromatographic
peakswere evaluatedto determinepeak purity. For
DMP it wasgenerallynecessary
to usepeakUV spectrato distinguishDMP from other
compoundseluting nearthe retentiontime of DMP.
HPLC calibration curvesobtained for BBP, DBP, and DEHP were found to be linear
overthe concentration
rangeof 0.001 mg/ml to 1 mg/ml. The calibrationcurvefor DMP
was linear from 0.001 mg/ml to 0.3 mg/ml, while DEP waslinear from 0.001 mg/ml
to 0.6 mg/ml. All regression
correlationcoefficients
werebetter than 0.995. The limit
of quantitation(LOQ) rangedfrom 1 to 10 ppm at ten times baselinenoise.Figure 1
showsthe chromatographic
separation
of the five phthalateestersand typicalchromatograms of cosmeticextracts.
The presenceof phthalateestersin solvents,laboratoryequipment,and plasticmaterials
hasbeenreportedby other investigators.
To assureaccuratequantitationof phthalate
estersin the cosmeticproductsexamined,laboratoryequipment and glasswarewere
carefullywashedand thoroughlyrinsedwith water and ethanolbeforeuse.The HPLC
PHTHALATE ESTERS IN COSMETIC PRODUCTS
131
mAU{
3000•
Standard
DEP
2500-:
BBP
2000-
DBP
1500 1000
50000
mAU
(b)Nail Enam .q
2500
DMP
2000
1500
1000
500
0
='0
3b
gi.
mAU
(c) Fragrance
3000
DEP
2500
2000
15001000-
00
mAU
3ooo(d) NailEnamel
DBP
2000
1500
00
Figure1. HPLCchromatograms
of(a)phthalate
ester
standards,
(b)nailenamel,
(c)fragrance
product,
and
(d) nail enamel.
132
JOURNAL OF COSMETIC SCIENCE
systemwas flushedwith mobile phaseat the beginningof eachday, and after each
injection the syringewas thoroughlywashedwith ethanol.SinceDEHP is particularly
persistentin the environment,and is widely usedasa plasticizer,plasticmaterialswere
not usedto process
samples.Solventblankswererun to confirmthe absence
of phthalares.No chromatographic
responses
were observedat the retentiontimes of any of the
phthalate esters.
Sincea widerangeof phthalateesterlevelsarepresentin cosmetic
products,thequantity
of sampleextractedwasoccasionally
varied,dependingon the expectedlevelof phthalate
esterin the product.For cosmeticproductshavingan unknownconcentration
of phthalate ester, a preliminary analysiswas made to determine the approximatelevel and to
confirmthe absenceof significantchromatographic
interference.
Then an appropriate
samplesizewasselectedfor analysis.For mostproducts1 g of samplewasanalyzed.
Method accuracy
wasevaluatedby performingrecoveryexperimentsfrom two hairspray
products,a handlotion, and an antiperspirant.Eachproductwasfortifiedwith the five
phthalate estersat levels of 100 and 1000 ppm. The resultsare shown in Table I.
Recoveries
of the five phthalateestersrangedfrom 73% to 112%. Averagerecoveries
for
DMP, DEP, BBP, DBP, andDEHP in the fourproductswere91%, 95%, 101%, 101%,
and 92%, respectively.
A surveyof a variety of consumercosmeticproductsfor phthalateesterswasconducted.
Productswerepurchased
from localstoresin the Washington,DC, area,includinghair
care products,deodorants,lotions and creams,nail products,fragrances,and body
washes.An attempt wasmadeto purchasemany of the sameproductsanalyzedby the
EWG (15) to confirmthe reportedphthalateesterlevels.The resultsof the analysisof
48 cosmetic
productsis shownin TableII. Levelslessthan 10 ppm arereportedasnot
detected.Sixty-sevenpercentof the productsanalyzedcontainedat leastonephthalate
ester,while hair sprays,deodorants,nail products,and hair moussecontainedtwo or
morephthalates.The highestphthalateesterconcentrations
werefoundin nail products,
with levelsobservedup to 59,815 ppm. DEP was the most commonphthalateester
found;it waspresentin twenty-seven
products.DBP wasfoundin ten products,while
DEHP wasnot foundin any producttested.With few exceptions,
therewasvery good
agreementbetweenthe phthalateesterlevelsfound and thosereportedby the EWG.
Differencesobservedmay be due to lot variations.
Table
I
Recoveryof PhthalateEstersfrom CosmeticProducts*
Percent recovery
Product
Fortificationlevel (ppm)
DMP
DEP
BBP
DBP
DEHP
HairsprayA
100
90
99
99
95
84
Hand lotion
1000
100
99
89
103
88
109
97
112
102
108
73
102
Antiperspirant
1000
92
94
95
95
100
83
94
100
103
88
95
100
99
1000
HairsprayB
100
1000
* Single determinationat eachlevel.
81
101
89
90
105
105
84
94
98
103
100
103
PHTHALATE
ESTERS IN COSMETIC
Table
PRODUCTS
133
II
Phthalate Esters in Thirteen Commercial Cosmetic Products (PPM)
Product
products
DMP•
Bodylotion
1
ND6
Hairspray
Deodorant
8
9
ND
ND
Fragrance
5
ND
DEP•
142
81, 118, 178, 204
38, 56, 57, 111,
681, 805, 2933
5486, 8851, 9081,
BBPa
DBP4
DEHP >
ND
ND
ND
43
ND
16, 38, 54
104
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
31, 43
ND
ND
ND
1•124, 38663
Skin lotion
3
ND
Hair gel
Hair mousse
Body wash
Shampoo
5
5
3
1
ND
ND
ND
ND
Hand cream
2
Nail enamel
6
84
53, 67
31, 56, 75, 128
200, 325
ND
ND
58, 143, 15395
27
1136
ND
107
ND
ND
ND
ND
ND
ND
25, 742,
46463, 59815
ND
Dimethylphthalate.
Diethylphthalate.
Benzylbutyl
phthalate.
Dibutylphthalate.
Diethylhexyl
phthalate.
Nonedetected
(<10ppm).
Numberoœ
products
containing
thephthalate.
The sourceof phthalateestersin most cosmeticproductsis most likely the fragrance
ingredient.Phthalateesterswereonly includedon the ingredientstatementsof someof
the nail products included in this survey.Individual fragranceingredientsare not
requiredto be includedin cosmeticproductlabeling(64).
The CosmeticIngredientReview(CIR) Expert Panel,an independentpanelof scientists
that hasbeenreviewingthe safetyof cosmeticrawmaterialssince1976, hasreviewedthe
safetyof severalphthalateestersusedin cosmeticproducts.In the first review,conducted
in 1985, the CIR concludedthat DMP, DBP, and DEP were safein cosmeticproducts
at levelsup to 5%, 25%, and 50%, respectively
(65). In a separatereviewof the safety
of BBP, the CIR concludedthat BBP is safe at concentrationsless than 1% (66). In
2003, the CIR rereviewedthe safetyof phthalateestersin cosmeticproductsin light of
reportsof phthalatemetabolitesin humanurine,andaffirmedtheir originalconclusions
that the levelsusedin cosmeticproductsweresafe.From 1998 to 2000, an expertpanel
convenedby the NTP concludedthat reproductiverisks from exposureto phthalate
esterswere minimal to negligiblein mostcases(67). The NTP hasconcludedthat food
is the primary sourceof human exposureto DBP (33). In the EuropeanUnion, the
ScientificCommitteeon Cosmetics
and Non-FoodProductsreviewedthe safetyof DEP
and concludedit wassafein cosmeticproducts(68); however,the committeeconcluded
that DBP shouldnot be intentionallyaddedto cosmetics(69).
The significance
of phthalateesterexposure
from cosmetics
comparedto exposure
from
food,water, air, and plasticmaterialsis difficult to assess.
Exposurefrom pharmaceuticalsmust alsobe factoredin, sincehigh urinary levelsof the metaboliteof DBP have
134
JOURNAL OF COSMETIC SCIENCE
been tracedto the useof drugs (70). Our surveyof cosmeticproductsfound that the
highestlevelsof phthalateesterswere presentin nail and fragranceproducts.Products
suchas nail polish hardenrapidly after application,and sophthalateesterabsorption
throughthe nail is likely to be significantlyinhibited. Exposureto phthalateestersfrom
productssuchas soaps,shampoos,
and conditionersthat are washedoff the skin soon
after applicationwill alsobe very low, due to limited contacttime with the skin. For
cosmeticproductsthat are left on the skin, exposureis a functionof the areaof skin
exposedto the productand the absorptionrate, and it hasbeenshownthat phthalate
esterabsorptionratesthroughhumanskin are slowcomparedto thoseof rodents(71).
Sinceour 2002 surveywasconducted,the FDA hasobservedthat somecosmeticproducts are being reformulatedto removephthalate esters.The FDA will continue to
monitor and evaluateall availabledata to assurethat phthalateesterlevelsin cosmetic
productsare not a health concern.The FederalFood, Drug and CosmeticAct doesnot
providefor premarketapprovalof cosmeticproducts,and the standardfor regulatory
action requiresthat the agencyprovea productis adulteratedor harmful under conditions of use.Basedon the safetyand toxicity data currentlyavailable,the agencyhas
concludedthat thereis no basisuponwhich to take regulatoryactionat this time. If the
FDA determinesthat a health hazardexists,the agencywill advisethe public and will
considerits regulatoryoptions.
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ESTERS IN COSMETIC
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