Final Report of the Safety Assessment of Alcohol Denat., Including SD Alcohol 3-A, SD Alcohol 30 ...

44 Pages·2007·1.18 MB·English

InternationalJournalofToxicology,27(Suppl. l):1—43,2008
Copyright©AmericanCollegeofToxicology
ISSN: 1091-5818print!1092-874Xonline
DOt: 10.1080,00915810802032388
Final Report of the Safety Assessment of Alcohol Denat.,
Including SD Alcohol 3-A, SD Alcohol 30, SD Alcohol 39,
SD Alcohol 39-B, SD Alcohol 39-C, SD Alcohol 40, SD
Alcohol 40-B, and SD Alcohol 40-C, and the Denaturants,
Quassin, Brucine Sulfate/Brucine, and Denatonium
Benzoate’
was 12.0 mg/kg. Brucine was nonmutagenic in an Ames assay at
Alcohol Denat. is the generic term used by the cosmetics in levels up to 6666 .sg/plate, with and without metabolic activation.
dustryto describe denatured alcohol. Alcohol Denat. and various In a repeat-insult patch test, for a hair care product containing
speciallydenatured(SD) alcoholsareusedascosmeticingredients 47% SD Alcohol 40 (95%), it was reported that Brucine Sulfate
inawide varietyofproducts. Manydenaturantshavebeen previ may be considered a nonprimary irritantand a nonprimary sen
ouslyconsidered,onanindividualbasis,ascosmeticingredientsby sitizer.Threedifferentsunscreenproducts(35% SDAlcohol40-B,
theCosmeticIngredientReview(CIR)ExpertPanel,whereasoth 72.4% SDAlcohol40,and74.5% SDAlcohol40)didnotshowany
ers,includingBrucineandBrucineSulfate,DenatoniumBenzoate, signsofphotoallergyinhumansubjects.Also,thesethreeformulas
andQuassin, have notpreviouslybeenevaluated. Quassinisabit did not exhibit any evidence ofphototoxicity in humans. Denato
teralkaloidobtainedfromthewoodofQuassiaarnara.Quassinhas niwnBenzoate is a bitter substance detectable at a concentration
beenusedasaninsectantifeedantandinsecticideandseveralstud of 10 ppb, discernibly bitter at 50 ppb, and unpleasantly bitter
ies demonstrate its effectiveness. At oral doses up to 1000 mg/kg at 10 ppm. The distribution oftopically applied lidocaine,a topi
using rats, Quassin was not toxic in acute and short-term tests, calanestheticchemicallyrelated to Denatonium Benzoatedemon
but some reversible piloerection, decrease in motor activity, and stratedthatvirtuallynolidocaineappearsintheplasma,suggesting
a partial loss ofrighting reflex were found in mice at 500 mg/kg. thatthelargerDenatoniumBenzoatemoleculealsowouldhavelit
At 1000 mg/kg given intraperitoneally (i.p.), all mice died within tleor nosystemicexposure. DenatoniumBenzoate (0.1%) did not
24hofreceivingtreatment.Inacytotoxicitytestwithbrineshrimp, show adverseeffects in 10 rats in an acute inhalationtoxicity test
1 mg/mIofQuassindidnotpossessanycytotoxicorantiplasmodial and 0.005% to 0.05% was nonirritatingto ocular mucosain 6 al
activity. Quassin administered to rat Leydig cells in vitro at con bino rabbits.TheacuteoralL5D0forthemaleratswas640mg/kg
centrations of5—25 ng/ml inhibited both the basal and luteinizing and for females, 584 mg/kg. The E5D0 for the male rabbits was
hormone (LH)-stimulated testosterone secretion in a dose-related 508 mg/kg and for the female rabbits, 640 mg/kg. In two chronic
fashion.Quassinatdosesupto2.0g/kgindrinkingwaterusingrats toxicity studies, Denatonium Benzoate was administered (by gay-
produced no significant effect on the body weights, butthe mean age) at 1.6, 8, and 16 mg/kg/day,one usingcynomologus monkeys
weightsofthe testes, seminal vesicles, and epididymidesweresig and the other rats, resulted in no compound-related toxicity. The
nificantlyreduced,andtheweightsoftheanteriorpituitaryglands toxicity ofSD Alcohols hasalsobeen tested, with implications for
were significantly increased. The sperm counts and levels ofLH, the particular denaturant used. An irritation test of55.65% SD
follicle-stimulatinghormone (FSH), and testosterone were signifi Alcohol 40-B denatured with Denatonium Benzoateusing rabbits
cantlyloweringroupstreatedwithQuassin.Brucineisaderivative produced minimal effects. A spray formula containing 12% SD
of2-hydroxystrychnine.Swiss-WebstermicegivenBrucinebase,30 Alcohol 40-B was found to be nonirritating when evaluated for
mi/kg, had anacuteoral L5D0 of150mg/kg, with central nervous vaginalmucosalirritationinNewZealandwhiterabbits.Cosmetic
system depression followed by convulsions and seizures in some formulationscontainingSDAlcohol40-B (denaturedwithDenato
cases.Inthoseanimalsthatdied,respiratoryarrestwasthecause. niuni Benzoate)werenotsensitizersin repeatedinsultpatchtests.
The acute Lp. E5D0 for 15 mI/kg ofBrucine basewas 62.0 mg/kg, A gel formula containing 29% SD Alcohol 40-B and a spray liq
withcentralnervoussystemdepressionpriortotheonsetofconvul uid containing 12% SD Alcohol40-B didnot inducephotoallergy,
sions,just as with oral Brucine. The acute intravenous (i.v.) L5D0 dermal sensitization, or phototoxic response in human subjects.
Although the absorption ofethanol (aka Alcohol for purposes of
cosmeticingredientlabeling)occursthroughskin,ethanoldoesnot
appear to affect the integrity ofthe skin barrier nor reach a very
Received25July 2007; accepted 14November2007. high systemic concentration following dermal exposure. Ethanol
‘Reviewedby the Cosmetic Ingredient Review Expert Panel. Ad maybefoundinthebloodstreamasaresultofinhalationexposure
dress correspondence to F. Alan Andersen, Cosmetic Ingredient Re and ingestion. Topically applied, ethanol can act as a penetration
view, 1101 17thStreet,NW,Suite412, Washington, DC20036,USA. enhancer. Most of the systemic toxicity of ethanol appears to be
2 COSMETIC INGREDIENTREVIEW
associatedwithchronicabuseofalcohol.Althoughethanolisdena This reportaddresses Alcohol Denat. andthe following spe
tured tomakeit unfitforconsumption, therehave been reportsof cific denaturedAlcohols: SD Alcohol 3-A, SD Alcohol 30, SD
intentional and unintentionalconsumption ofproductscontaining Alcohol 39, SD Alcohol 39-B, SD Alcohol 39-C, SD Alcohol
denatured alcohol. Ethanol is a reproductive and developmental
40, SDAlcohol40-B, andSDAlcohol40-C. Thesearethemost
toxicant. Ethanol is genotoxic insometestsystems and ithasbeen
proposed thatthe genotoxic effects ofethanolare mediated via its widely used Alcohol Denat. ingredients in cosmetics.
metabolite, acetaldehyde. A briefsummary is provided ofthe ef Denaturants in Alcohol Denat. and the above SD Alcohols
fects of chronic ingestion of alcohol including intoxication, liver include many ingredients previously reviewed by the Cosmetic
damage, brain damage,and possiblecarcinogenicity.The CIREx Ingredient Review (CIR) Expert Panel and will not be exten
pert Panel recognizes that certain ingredients in this group are
sively discussed. In addition to the many denaturants already
reportedlyused in agivenproductcategory,buttheconcentration
of use is not available. Because dermal application or inhalation considered,Quassin,BrucineSulfate,Brucine,andDenatonium
ofcosmeticproductscontainingtheseingredientswillnotproduce Benzoate are used and are afocus ofthis review.
significantsystemicexposuretoethanol,theCIRExpertPanelcon PartIofthis safety assessment addressestheavailable safety
cluded that safety ofthe ingredients should be predicated on the testdatarelevanttothese denaturants in AlcoholDenat. and the
safety ofthe denaturants used. The Panel considered that the ad
widely used SD Alcohols.
verseeffectsknowntobeassociatedwithAlcoholingestionincluded
BecauseethanolistheprimarycomponentofAlcoholDenat.
in this safety assessment do notsuggest a concern for Alcohol De
nat. or SD Alcohols because of the presence of the denaturants, andtheSDAlcohols,PartIIisanoverviewofsafetytestdataon
which are added for the express purpose of making the Alcohol ethanolitself. Othertermscommonlyusedincludeundenatured
unpotable.TheCIR ExpertPanel haspreviouslyconductedsafety ethyl alcohol orjustethyl alcohol.
assessmentsoft-ButylAlcohol,DiethylPhthalate,MethylAlcohol,
In concept, the entire purpose of denaturing Alcohol is to
SalicylicAcid, Sodium Salicylate,and Methyl Salicylate,in which
each was affirmed safe orsafewith qualifications. Given their use makeitunfittodrink, sothe safety assessmentdoes notinclude
asdenaturantsareatlowconcentrationsofuseinAlcohol,the CIR the vast literature on alcoholism. On the other hand, Alcohol
Expert Panel determined that Alcohol Denat. denatured with t Denat.maybeingested, soalimitednumberofreferencestothe
Butyl Alcohol, Diethyl Phthalate, Methyl Alcohol, Salicylic Acid, effects ofAlcohol ingestion are included.
Sodium Salicylate, and Methyl Salicylate is safe as used in cos
metic formulations with no qualifications. Likewise, because they
are denatured with either t-Butyl Alcohol, Diethyl Phthalate, or PART I: DENATURANTS
MethylAlcohol, SD Alcohols3.A, 30,39-B, 39-C, and40-C all are
Denatured alcohols, in the United States, must conform to
considered safe as used. The Panel considered the available data
for Denatonium Benzoate and SD Alcohol 40.B to be sufficient to the Code ofFederalRegulations (CFR) specifications forcom
support the safety of these ingredients in cosmetics. Denatonium pletely and specially denatured alcohol found in 27CFR2l.
Benzoate is sufficiently bitter that it is an effective denaturant at Table 1 lists the specifications for those denatured alcohols in
only0.0006%.The Panel recognizedthatdataondermal penetra thissafetyassessment—thosedenaturantsinboldtypehavebeen
tion of Denatonium Benzoate were not available, but considered
previouslyconsideredbytheCIRExpertPanel,withthefollow
thatthe available data on lidocaine, a smallerstructurally related
chemical,indicatesthatdermalexposuredoesnotresultinmeasur ing conclusions:
ablesystemicexposure. Theavailabledata, however,were notsuf
ficienttosupportthesafetyofQuassin, Brucine,and Brucine Sul t-Butyl Alcohol—safe as used in cosmetic products (Andersen
fate, Alcohol Denat. denatured with those denaturants, or SD Al 2005a);
cohol39and SD Alcohol40(SDAlcoholsdenaturedwithQuassin, Diethyl Phthalate—safe for topical application in the present
Brucine,and/orBrucineSulfate),andinorderfortheExpertPanel practices of use and concentration in cosmetics (Elder
to reach a conclusion for these denaturants, additional data are
1985), reaffirmed safe for use in the present practices of
needed.
use and concentration (Andersen 2005b);
Methyl Alcohol—safe as used to denature alcohol used in cos
metic products (Andersen 2001); and
Salicylic Acid, Sodium Salicylate, andMethyl Salicylate—safe
asusedwhenformulatedtoavoidirritationandwhenformu
INTRODUCTION latedto avoidincreasingsun sensitivity, or,when increased
According to the International Cosmetic IngredientDictio sunsensitivitywouldbeexpected,directionsforuseinclude
naryandHandbook(Gottschalck and McEwen2004), Alcohol the daily use ofsunprotection (Andersen 2003).
is the name given by convention to ethanol as used in cosmet
ics, and Alcohol Denat. is the name given to denatured Alco The denaturants noted above will notbe discussed furtherin
hol foruse in cosmetics. Alcohol Denat. is prepared by adding this safetyassessment. The safetyofAlcoholDenat. andtheSD
one or more denaturing agents to potable Alcohol, making it Alcohols containingthese ingredients may be inferred.
unfit for beverage or intemal human medicinal use. Alcohol Theremainderofthis sectionwill focus onthreedenaturants
may be completely denatured (CD Alcohol) or specially dena listed in the International Cosmetic IngredientDictionaiy and
tured (SD Alcohol) depending on the type(s) of denaturant(s) Handbook (Gottschalck and McEwen 2004), Quassin, Brucine
used. Sulfate, and Denatonium Benzoate, which have not previously
ALCOHOLDENAT. 3
TABLE 1
Specifications inthe Code ofFederalRegulations (27CFR21) forspecially DenaturedAlcohol
Ingredient 100 Gallons ofEthanol DenaturedWith
SD Alcohol 3-A 5 gallons ofmethyl alcohola
SD Alcohol 30 10 gallons ofmethyl alcohol”
SD Alcohol 39 9 pounds ofsodium salicylate”, U.S.P., or salicylic acid”, U.S.P., and 1.25 gallons offluidextract ofquassia,
N.F., and 1/8th gallon oft-butyl alcohol”
SD Alcohol 39-B 1/8th gallon oft-butyl alcohol” and2.5 gallons ofdiethylphthalate”
SD Alcohol 39-C 1 gallonofdiethylphthalate”
SD Alcohol 40 1/8th gallon oft-butyl alcohol” and 1.5 avoirdupois ounces ofanycombinationofone ormore ofthe
following: brucine (alkaloid),brucine sulfate, N.F., orquassin
SD Alcohol 40-B 1/16th avoirdupois ounceofdenatoniumbenzoate,N.F., and 1/8th gallonoft-butyl alcohol”
SD Alcohol40-C 3 gallons oft-butyl alcohol”
aDenaturantsforwhich CIRhascompletedasafetyassessment seetext.
-
beenaddressed.Brucineisalsoincludedbecauseitisspecifically it absorbs light at 255 nm (Budavari 2001). The calculated
listedas an approveddenaturantinTable 1. octonol/waterpartitioncoefficientforQuassinis0.94(Syracuse
Research Corp. 2005).
CHEMISTRY According to Hunt (1967), Quassinhas been used for many
years as adenaturantforalcoholusedincosmeticpreparations.
Definition and Structure Itisodorless and extremelybitter. Quassinis soluble in chloro
Quassin form, ethyl acetate, andbenzene, andsparingly soluble inether
According to the International Cosmetic IngredientDictio or lightpetroleum. The concentration ofQuassin may vary ac
naryandHandbook(GottschalkandMcEwen2004),Quassinis cording tothe differentregulations governingdifferenttypes of
abitteralkaloidobtainedfromthewoodofQuassiaan1ara,CAS products as well as the alcohol content.
number76-78-8 (the CAS number ofd, l-quassin is 75991-65-
0). Othernames forQuassin include Brucine andBrucine Sulfate
Brucine Sulfate is an alkaloid thatconforms to the formula:
2,12-dimethoxypicrasa-2,12-diene-1,11,16-trione; and (2C3H2N204)2.2S4720HHO6 (CAS number4845-99-2) with a
Picrasa-2,12-diene-1,11,16-trione,2,12-dimethoxy-. molecular weight of 394.46 (Budavari 1989). Brucine Sulfate
currently islisted as adenaturantintheInternationalCosmetic
StojanacandValenta(1991)reportedthatd, l-quassincanbe
synthesizedviaa 13-stepprocess startingwithesterhemiacetal. IngredientDictionaryandHandbook(GottschalckandMcEwen
The molecular formula of Quassin is C2H70678 (molecu 2004), with the followingtechnical/othernames:
lar weight is 388.45) and the structure is shown in Figure 1 • Brucine Sulfate Heptahydrate;
(Budavari 1989). The melting point of Quassin is 222°C and • 2,3-dimethoxystrychnidin-lO-oneSulfate (2:1);
o Strychnidin-10-one,2,3-dimethoxy-,Sulfate(2:1); and
O3CH • Strychnidin-10-one,2,3-dimethoxy-, Sulfate (1:1),
Heptahydrate.
.3CH
The CAS numberofBrucine is 357-57-3. Brucine currently
isnotlistedintheInternationalCosmeticIngredientDictionary
a/3dHandbookasacosmeticingredient.Thisalkaloidresembles
strychnine, with the formula C2H6N204,3, a molecular weight
of 304.46, absorption maxima at 263 and 301 nm, a melting
point of 178°C, apH (saturated water solution) of9.5, and the
structure shown inFigure 2 (Budavari2001).
Thecalculatedoctonol/waterpartitioncoefficientforBrucine
is 1.49 (SyracuseResearch Corp 2005).
ArenaandDrew(1986)notedthatBrucineSulfateisintensely
bitter, but they suggest that quantities present in denatured al
FIGURE 1 cohol do not create any greathazard, unless an unusually large
StructureofQuassin(Budavari 1989). quantity is ingested.
4 COSMETICINGREDIENTREVIEW
FIGURE 2
StructureofBrucine (Budavari 1989).
DenatoniumBenzoate
AsgivenintheInternationalCosmeticIngredientDictionary
andHandbook (Gottschalck and McEwen 2004), Denatonium
Benzoate (CAS number 3734-33-6) is an organic quaternary
ammonium compound that functions as a denaturant and con
formstothefollowingempiricalformula: C2H2N207H502.•19C
The structure of Denatonium Benzoate is shown in Figure 3.
Technical/othernames include:
• Benzenemethanaminium, N-[2-[(2,6-dimethylphenyl)
amino]-2-oxoethyl]-N,N-diethyl-,benzoate;
• N-[2-[(2,6-dimethylphenyl)aminol-2-oxoethyl]-N,N-
diethylbenzenementhanaminiumbenzoate (RIFM);
FIGURE 3
• N-[2-[(2,6-dimethylphenyl)amino]-2-oxoethyl]-N,N- StructureofDenatoniumBenzoate(GottschalckandMcEwen2004).
diethylbenzene-methanaminium benzoate (RIFM);
and mended concentration torenderaformulation aversivelybitter;
• N-[2-[(2,6-dimethylphenyl)aminol-2-oxoethyl]-N,N- however, up to 100 ppm has beenused (Hendersenet al. 1998;
diethylbenzene-methanaminiumbenzoate. Klein-Schwartz 1991; Payne 1989).
As shown in Table 1, SD Alcohol 40-B is denatured with
Denatonium Benzoate has a molecular weight of 446.58
1/16 avoirdupois ounce ofDenatonium Benzoate,N.F., and 1/8
(Budavari 1989). It is available as a white, odorless granule.
gallonoftert-butylalcohol.AsdescribedbyEquistarChemicals,
The melting point ranges from 163°C to 170°C. Denatonium
LP (2004), this results in SD Alcohol40-B with approximately
Benzoateis soluble in waterand alcohol, somewhatin acetone,
0.0006% Denatonium Benzoate and 0.12% t-butyl alcohol.
and nearly insoluble inether (Hansen et al. 1993). Denatonium
The CIR Expert Panel also considered severalpieces ofdata
Benzoate absorbs radiation at 263 nm (at a concentration of
on lidocaine, a topical anesthetic structurally related to Dena
100 JLg/ml in water) and the pH of a 3% solution in water is
tonium Benzoate. AUnited States Patent (Hay 1963) provided
between 6.5 and 7.5 (Committee ofRevision ofthe U.S. Phar
thechemicalstructureofaquatemarycarboxylate derivedfrom
macopeialConvention2004). Thecalculated octonol/waterpar
lignocaine, a synonymforlidocaine.
tition coefficient for Denatonium Benzoate is 1.78 (Syracuse
The structure oflignocaine (aka lidocaine) is represented in
Research Corp. 2005).
Figure4 and may be compared to the structure ofDenatonium
According to Jackson and Payne (1995), Denatonium Ben
BenzoateinFigure3.Lignocainebenzylbenzoateandlidocaine
zoate is one ofthe most bitter substances. It is added to a wide
benzyl benzoate are synonyms for Denatonium Benzoate (Hay
variety ofproducts atrequired low levels, but does not interfere
1963).
with the mode ofaction ofthe product. It is detectable by taste
atconcentrations as low as 10 ppb, discernibly bitter at 50 ppb,
and at 10ppm itis unpleasantly bitter. Method of Manufacture
For the purposes of denaturing alcohol, Denatonium Ben Information on specific CD or SD Alcohols was not found.
zoate is added at a rate of 6 ppm in the USA and 10 ppm in The CIRExpertPanelpresumedthatTable 1 describesasimple
the UK. In most other applications’ 20 to 50 ppm is the recom mixing process.
ALCOHOLDENAT.
Analytical Methods
Quassin
Hunt (1967) described a method by which Quassin may be
determinedwhenused asadenaturantforthe alcohol contained
incosmeticproducts.Thetwobitterprinciples, neoquassinand
Quassin, are determined together as Quassin, after oxidation
withasolutionofsodiumdichromateinglacial aceticacid.Per
fumeoilsarefirstremovedbysolventextractionandtheaqueous
phase is then evaporated to dryness and the residue oxidized.
Further solvent-extraction steps yield a solution ofQuassin in
chloroform, which is applied to a thin-layer chromatographic
plate. The spots arecomparedvisually with standardsprepared
from an oxidized “quassin” solution to permit determinations
withintherange of5 to 36ppm in samples.
Brucine
FIGURE 4 Sondack and Koch (1973) noted that there are many meth
Structureoflignocaine,akalidocaine(Hay 1963). ods in which Brucine and strychnine are estimated, includ
Quassin inggravimetricandtitrimetricanalyses,paperchromatography,
Clark (1937) stated that quassin is prepared by extracting thin-layer chromatography (TLC), ion-exchange chromatogra
Quassia chips (20 kg) and allowing themto stand inhotwater phy,spectrophotometry,andnuclearmagneticresonance(NMR)
for 3 h. The extract is poured off and the process is repeated spectroscopy. Brucine and strychnine have been separated by
three times. The extractis treated with solutions ofleadacetate gas—liquid chromatrography (GLC).
andcarbon (Carbex)toabsorbthe Quassin.The amountoflead Zhang et al. (2002) reported a new capillary electrophoresis
acetateandcarbonthatisaddedisinaratioof4to 1.Thecarbon procedure with field-enhanced stacking concentration for the
isremoved, air-dried,andmixedwithchloroform.Themixtures analysis of strychnine and Brucine. These two alkaloids can
are concentrated to dryness under reduced pressure, dissolved be separated after the optimization ofthe separation and con
in 150 cc of methanol, and mixed with 950 cc of hot water. centration within 5 mm andquantifiedforhigh sensitivity. The
A crystallization of crude Quassin begins once the mixture is detectionlimitsforstrychnineandBrucinewere 1.0ngm11 and
filtered. The crystallization is complete within2days. 1.4ngml’,respectively. Themethodwasusefulforqualitative
and quantitative analysis of strychnine and Brucine in Strych
Brucine nos nux-vomica L with recovery of 105.1% for strychnine and
Tedeschietal.(1968)reportedthatBrucine(2,3-dihydroxyst- 98.4% forBrucine.
rychnine) ispreparedbyreacting 2-hydroxystrychnineinacetyl
alcohol with a solution ofFremy salt in water. This reaction is DenatoniunzBenzoate
allowed to complete, and 2 N H4CIO and N2S2O4a are added. The concentration of Denatonium Benzoate was measured
ThepHisadjustedwithN4OHHto7to7.5.Thismixtureisthen by high-pressure liquid chromatography (HPLC). The analysis
dissolved in C3/MHetChy1l Alcohol and treated with ethereal of standards showed a linear curve in the range of 1.25 to 50
diazomethane.Thetwoproductsthatemergeare-colubrinand ppm (Henderson etal. 1998).
Brucine.
DamonandPettitt(1980)reportedamethodforthedetermi
According to the MerckIndex, Brucine also can be isolated nationofDenatoniumBenzoateinrapeseedoil.Themethoduti
from Strychnos seeds (Budavari 2001). lizes a simple extraction and concentration technique followed
DenatoniurnBenzoate byreverse-phasehighperformanceliquidchromatography.The
Lignocaine benzyl benzoate (a synonym for Denatonium columneluentwasmonitoredat210nmandpeakareadatawere
Benzoate) is describedbyHay (1963) in aUnited StatesPatent generated by computing integrator. Detection was possible be
describing the use of this quaternary carboxylate as a denatu low 5 ngandcalibrationcurves werelinearto 100ngormore.
rant for Ethyl Alcohol. The patent presents multiple synthetic
approaches to the preparation of Denatonium Benzoate. The Impurities
patent states thatthe method ofdenaturingEthylAlcoholcom Brucine
prises adding 0.01% to 0.001% Denatonium Benzoate (note: Brucinealkaloid(27CFR21.99)mustpasstwotestsinorder
as described earlier, 1/16 avoirdupois ounceper 100 gallons of forits usetobe authorized: strychninetestand sulfatetest. The
Ethyl Alcohol is the currently approved level of Denatonium strychnineteststatesthatBrucinealkaloidshallbefreeofstrych
Benzoate as given inthe CodeofFederal Regulations). nine when tested by the method listed under Brucine Sulfate,
6 COSMETIC INGREDIENTREVIEW
N.F. IX. Ifthe Brucine contains as much as 0.05% strychnine, Forexample,AlcoholDenat.wasreportedtobeusedin9out
acleardistinctiveviolet color, characteristic ofstrychnine, will ofatotal of215 bubblebaths (4.2%) ataconcentrationof0.1%,
be obtained. The sulfate test states that no white precipitate is but the denaturants used were not reported: whereas Alcohol
formed that is not dissolvedby hydrochloric acid when several Denat. wasreportedtobeused in264 of684colognesandtoilet
drops ofa 1 Nbarium chloride solution are addedto 10ml ofa waters (38.6%)atconcentrationsfrom79%to96%,withdiethyl
solution ofthe alkaloid. phthalate, t-butyl alcohol, and Denatonium Benzoate given as
examples ofdenaturants used in this category (althoughnot to
Quassin getherin one product ornecessarily in allproducts).
Quassin (27CFR21.l24) mustpass twotests inorderforits For each SD alcohol in Table 3 for which there is a formula
use to be authorized: optical assay and solubility. The optical for denaturants in Table 1, the denaturants used are given next
assay states that when 1 g of Quassin (in solution in a small to the name. For example, SD Alcohol 3-A (denatured with
amount of95% alcohol) is dissolved in 10,000 ml ofwater, the Methyl Alcohol only) is used in atotal of 16 cosmetic products
absorbanceofthe solutionina 1-cmcell atawavelengthof258 at a concentrationrangefrom0.1% to 5%.
m shall not be less than 0.400. The solubility test states that Table 3 also includes ingredient use in categories for which
0.5 g of Quassin dissolves completely in 25 ml of 190 proof no reports were provided to FDA, but for which a currentcon
centration of use was provided by the Cosmetic, Toiletry and
alcohol.
Fragrance Association (CTFA), For example, Alcohol Denat.
wasnotreportedtoFDAtobeusedineye shadowproducts,but
DenatoniumBenzoate
industry reported that it is currently being used in this product
The Committee of Revision of the United States Pharma
category in the concentration range of0.008% to 6%.
copeial Convention (2004) states that Denatonium Benzoate,
DenaturingagentsareregulatedintheUnitedStates,asgiven
driedat 105°Cfor2h, containsonemoleculeofwaterofhydra
in Table I, and Europe. Alcohol Denat. is not included among
tion oris anhydrous. When driedat 105°Cfor2h, itcontainsno
the substances listed as prohibited, restricted, or provisionally
lessthan99.5%andnomorethan 101.0%o7H3N2O3.f84CWhen
allowed in the use of cosmetic products marketed in Europe
DenatoniumBenzoateisdriedat 105°Cfor2h,themonohydrate
(European Commission 1999) or in Japan (Ministry ofHealth,
loses between 3.5% and 4.5% ofits weight, and the anhydrous
Labor, and Welfare [MHLWI 2001a, 2001b). Brucine is listed
form loses no more than 1.0% of its weight. The residue on
inAnnexII(substances whichmustnotformpartofthecompo
ignitioncannotbemorethan0.1%.When900mgofDenatonium
sition ofcosmeticproducts) oftheEuropean Unionregulations
Benzoate is dissolved (dried and accurately weighed) in 50 ml
(European Commission 2005).
ofglacial acetic acid, 1 drop ofcrystal violet TS is added, and
titrated with 0.1 N perchloric acid VS to a green end point. A Noncosmetic
blankdeterminationisperformed,andanycorrectionsaremade. Alcohol Denat. is used in medical, industrial, and domestic
Eachmilliliterof0.1 Nperchloric acidisequivalentto44.66mg products. For example, methylated spirit contains 5% methyl
ofC2H3N203.84 alcohol in ethanol; Solox (Shellac solvent) contains 5% methyl
alcohol, 1% gasoline, 1% ethyl acetate, and 1%methylisobutyl
ketone in ethanol; Sterno-canned heat contains 3.7% methanol
USE
and 4% acetone in ethanol; and rectified spirit contains 90%
ethanol (Bastani et al. 1976). AlthoughFDA/CFSAN has listed
Cosmetic
Alcohol Denat. as one ofthe food additives in the “Everything
According to the International Cosmetic IngredientDictio
Added to Food in the United States” (EAFUS) list, there is no
nary andHandbook (Gottschalk and McEwen 2004), Alcohol
reporteduse ofit as afood additive (FDA, 2003).
Denat. functions as an antifoaming agent, cosmetic astringent,
solvent, and viscosity-decreasing agent in cosmetics, and all of GENERAL BIOLOGY
the SD alcohols function as cosmetic astringents, solvents, and
viscosity decreasing agents. Table 2 lists the authorized uses Absorption, Metabolism, and Distribution
for the denatured alcohols in the Code of Federal Regulations Quassin
(27CFR20 and 21). Garcia et al. (1997) reported that an intestinal transit test
AsreportedbyindustrytotheFoodandDrugAdministration was performed using Quassin extract. Thirty male mice were
(FDA),Table3 givesthereportedfrequencyofuseas afunction distributed into three equal groups. After 6 h of fasting, each
ofproduct category, along with the total number ofproducts in group was administered one of the following treatments: 0.5
thatcategory. Table 3 also includes currentconcentration ofuse ml of distilled water and Quassin extract in doses of 500 and
information provided to CIR directly by the industry. If a par 1000mg/kg. Thedatawererecorded andexpressedasthemean
ticulardenaturantwas reportedly used in thatproduct category, value±thestandarderror.Withthe500mg/kgdose,theaverage
it is listed (in bold type, if the denaturant has been considered percentageofintestinaltransitwas63.86%±7.15,whichdidnot
previously in aCIR safety assessment). constitute a statistically different value upon comparison with
ALCOHOLDENAT. 7
TABLE2
Authorized uses as a solvent for SD Alcohols inpersonal careproducts in the Code ofFederalRegulations (27CFR20and 21)
Ingredient Authorized Uses as a Solvent inPersonal CareProducts
SD Alcohol 3-A Shampoos, soap and bathpreparations
SD Alcohol 30 Soap andbath preparations
SD Alcohol 39 Hairand scalp preparations,bayrum, lotions and creams (hand, face, andbody),perfume and perfume
tinctures, toilet waters andcolognes, shampoos, soap andbathpreparations
SD Alcohol 39-B Hair and scalppreparations, bay rum, lotions and creams (hand, face, andbody), deodorants (body),
perfumes and perfumetinctures
SDAlcohol 39-C Hairand scalppreparations, lotions andcreams (hand, face, andbody), deodorants (body), perfumes and
perfumetinctures, toilet waters andcolognes, soaps andbath preparations
SDAlcohol 40 Hair andscalppreparations, bay rum, lotions andcreams (hand, face, andbody), deodorants (body),
perfumes and perfume tinctures, toiletwaters andcolognes, shampoos, soaps andbath preparations
SD Alcohol 40-B Hairand scalppreparations, bayrum, lotions andcreams (hand, face, andbody), deodorants (body),
perfumes and perfume tinctures, toilet waters andcolognes, shampoos, soaps andbath preparations
SD Alcohol40-C Hairand scalppreparations, bayrum, lotions andcreams (hand, face, andbody), deodorants (body),
perfumes andperfumetinctures,toiletwaters and colognes, shampoos, soaps andbath preparations
the control group (58.19% ± 8.82). The group that received activity test showed Quassin produced 90% to 100% damage
1000 mg/kghad an averagepercentintestinal transitof71.96% with all applications and on all days. Chlordimeform also pro
±4.96.
ducedthis result. Forthe insecticidal activitytest, protocol was
Lidocaine the exact same, in which Quassin produced potent insecticidal
activityaswell. Itwasconcludedthatthedeathofthelarvaewas
As seen in Figures 3 and 4, lidocaine and Denatonium not dueto any antifeedantactivity ofthecompoundstested,but
Benzoate are chemicallyrelated.
that these compounds had lethality.
Keams etal. (2003) used an iontophoreticdrugdelivery sys Parketal. (1987)reportedthatQuassin,among otherquassi
temtogivelidocaineto 12children(ages5 to 15). Thechildren noids, was tested upon acute insecticidal activity against the
wereexposedtolidocaine(100mgofa 10%solution)for0,0.3, American cockroach. Thequassinoids were subcutaneouslyin
and 3.5 h. Plasma levels oflidocaine, measured for up to 10 h jected into the cockroach and observations were made at 3, 24,
afterthistreatment,werebelow 10ng/mlatalltimepoints. This and 48 h with recordings ofthe minimum effective dose (,ug/g
method, which enhances entry into the skin, did not result in a of insect body weight) to cause ataxia. All of the quassinoids,
measurable systemic dose.
includingQuassin,hadstrongacuteparalyticactivityatdosages
CTFA (2005) offered the commentary that lidocaine is a as low as 25 iig/g of insectbody weight, but after24h most of
smaller molecule than is Denatonium Benzoate. Even with an theinsectsrecoveredandthespecificactivityfellto 100,ug/gof
iontophoretic drug delivery system, little of the lidocaine was insectbodyweightormore. Whenahigherdosagewasapplied,
found in plasma, suggesting that the larger Denatonium Ben acute insecticidal activity resulted and there was no recovery.
zoate,intheabsenceofadeliverysystemotherthanitspresence The authors noted that Quassinand neoquassin were noteffec
as an alcoholdenaturant, wouldbeunlikely toresultinany sys tive as an insecticide against the cockroach under a different
temic exposure.
evaluation method that was focused on delayedtoxicity.
Leskinen et al. (1984) reported in a study where 13 differ
Insect Antifeedant and Insecticidal Activities ent quassinoids were tested for antifeedant activity against the
Quassin
Mexican bean beetle (Epilachna varivestis Mulsant) and the
Daido et al. (1995) noted that many cytotoxic quassi southern armyworm(Spodopteraeridania). Quassin wastested
noids have been shown to have insect antifeedant activity. at500, 250, 100, 50, and 10ppm with the Mexicanbeanbeetle
The quassinoids were isolated from P.ailanthoides and the an and 500, 250, and 100 ppm with the southern armyworm. Five
tifeedant and insecticidal activities were measured against the trials ateach concentration were performed bycoating one leaf
diamondback moth, Plutella xylostella. These activities were with the compound and one with just a solvent. The subjects
comparedto aknown antifeedant, chiordimeform. were placed on the leaves, and after 24 h the percentage eaten
The quassinoid tested was applied to the cabbage leafdiscs for each leaf was estimated. In every test conducted with the
fedtothemoth at63.7,31.9, and 16.0 t2g/cm andobservations Mexican bean beetle, more ofthe control leafwas eaten com
were made on days 1, 2, and 5. The results ofthe antifeedant pared to the leafwith Quassin applied to it. When leaves eaten
8 COSMETICINGREDIENTREVIEW
TABLE 3
Alcohol Denat. and SD Alcohol use incosmetics
ProductCategory NumberofFormulations Use
(Total formulations in productcategory FDA 2002) Containing Ingredient Concentration
-
(Denaturant, ifknown a CTFA 2003) (FDA 2002) (CTFA 2003)
-
AlcoholDenat.
BathPreparations
Bath Oils, Tablets, and Salts (143) 6 0.005%
Bubble Baths (215) 9 0.1%
OtherBath Preparations (196) 0.07%
Eye Makeup Preparations
Eyeliner (548) (t-Butyl Alcohol; DenatoniumBenzoate) 3 2—10%
Eye Shadow (576) (DenatoniumBenzoate) 0.008—6%
—
Eye Makeup Remover (100) (Denatonium Benzoate) 3%
Mascara (195) (DenatoniumBenzoate) 3—7%
OtherEye Makeup Preparations (152) (Diethyl Phthalate; Denatonium Benzoate) 5—10%
Fragrance Preparatons
Colognes and Toilet Waters (684) (Diethyl Phthalate; t-Butyl Alcohol; 264 79—96%
Denatonium Benzoate)
Perfumes (235) (Diethyl Phthalate;t-ButylAlcohol; DenatoniumBenzoate) 79 70—97%
Dusting andTalcom Powders (273) 0.008%
Sachets (28) (t-Butyl Alcohol) 4 86%
OtherFragrancePreparations (173) (t-ButylAlcohol; DenatoniumBenzoate) 20 30—98%
Non-coloring Hair Preparations
HairConditioners (651) (Denatonium Benzoate) 9 0.1—40%
Hair Sprays (aerosol fixatives) (275) (DenatoniumBenzoate) 60 76—90%
Rinses (non-coloring) (42) 2
Shampoos (non-coloring) (884) (DenatoniumBenzoate) 11 0.1—25%
Tonics, Dressings, and Other HairGrooming Aids (598) (DenatoniumBenzoate) 53 35—91%
Wave Sets (53) (DenatoniumBenzoate) 6 89%
OtherHairPreparations (277) (Denatonium Benzoate) 27 29—83%
HairColoringPreparations
HairBleaches (120) 3
Other Hair Coloring Preparations (55) (DenatoniumBenzoate) 0.2—16%
Makeup Preparations
Blushers (245) 0.008%
Face Powders (305) 0.008%
Leg and Body Paints (4) 0.03%
—
Foundations (324) (t-Butyl Alcohol; DenatoniumBenzoate) 5 4—10%
OtherMakeup Preparations (201) (t-Butyl Alcohol; DenatoniumBenzoate) 1 7—12%
Nail CareProducts
Basecoats andUndercoats (44) 1 10%
Cuticle Softemers (19) 3%
Nail Polish and Enamel (123) 10—52%
NailPolish andEnamel Removers (36) 2
—
OtherManicuring Preparations (55) (Diethyl Phthalate; DenatoniumBenzoate) 4 3—48%
Oral HygieneProducts
Mouthwashes and BreathFresheners (46) 20
ALCOHOLDENAT. 9
TABLE3
AlcoholDenat. andSD Alcohol use incosmetics
ProductCategory
Number ofFormulations Use
(Total formulationsinproduct category FDA 2002) Containing Ingredient Concentration
(Denaturant, ifknowna - CTFA 2003) (FDA2002) (CTFA 2003)
Personal CleanlinessProducts
Bathsoaps and detergents (421)
0.2%
Deodorants (underarm) (247) (t-ButylAlcohol; DenatoniumBenzoate) —47 72—99%
Douches (5)
2
Feminine Deodorants (4)
1
OtherPersonal CleanlinessProducts (308) 17 9—82%
Shaving Preparations
AftershaveLotion (231) (Diethyl Phthalate; t-Butyl Alcohol) 77 55—97%
PreshaveLotions (alltypes) (14)
10 87%
Shaving Cream (134)
0.07%
OtherShaving PreparationProducts (63) (t-Butyl Alcohol; Denatonium 11 5—70%
Benzoate)
Skin Care Preparations
Skin Cleansing Products (coldcreams,lotions, liquids, andpads) (775) (Diethyl 15 0.7—51%
Phthalate andDenatoniumBenzoate; Denatonium Benzoate)
Depilatories (34) (DiethylPhthalate)
1%
Face andNeck skin care (excl. shaving) (310) (Diethyl Phthalate; Denatonium 3 2—80%
Benzoate)
Body and Handskincare (excl. shaving) (840) (Diethyl Phthalate andt-Butyl 28 2—40%
Alcohol; DiethylPhthalate; DenatoniumBenzoate)
FootPowders and Sprays (35) 4 0.01%
Moisturizers (905) (Diethyl Phthalate; DenatoniumBenzoate) 15 0.1—26%
Nightskin care (200) (Denatonium Benzoate) 1 0.005—25%
Paste Masks (mudpacks) (271) (DiethylPhthalate; DenatoniumBenzoate) 11 1—5%
SkinFresheners (184)
38 10%
OtherSkin CarePreparations (725) (Diethyl Phthalate; t-ButylAlcohol; 32 0.6—90%
DenatoniumBenzoate)
SuntanPreparations
SuntanGels, Creams, andLiquids (131) (Diethyl Phthalate; t-Butyl Alcohol) 3 2—74%
IndoorTanning Preparations (71)
0.0008%
OtherSuntanPreparations (38) (DenatoniumBenzoate) —1 50%
2002 total/range forAlcohol, Denat. 821 0.0008—99%
SDAlcohol3-A(denaturedwithMethylAlcoholonly)
BathPreparations
BubbleBaths (215)
5 4%
OtherBathPreparations (196) 3
Non-coloring HairPreparations
Shampoos (non-coloring) (884) 1 3%
PersonalCleanlinessProducts
Bath Soaps and Detergents (421)
5
Skin CarePreparations
SkinCleansing Products (coldcreams, lotions, liquids, and pads) (775) 2 5%
Skin Fresheners (184)
0.01—2%
2002 total/rangeforSD Alcohol 3-A —16 0.01—5%
SDAlcohol39-B (denaturedwith t-ButylAlcoholandDiethylPhthalateonly)
FragrancePreparations
Colognes andToiletWaters (684) 1
(Continuedon nextpage)
10 COSMETICINGREDIENTREVIEW
TABLE 3
Alcohol Denat. and SD Alcoholuse in cosmetics (Continnued)
Product Category Number ofFormulations Use
(Total formulations in productcategory FDA 2002) ContainingIngredient Concentration
-
(Denaturant, ifknown a CTFA 2003) (FDA 2002) (CTFA 2003)
-
Shaving Preparations
Other Shaving PreparationProducts (63)
1998 totals/ranges for SD Alcohol 39-B 2
SD Alcohol39-C(denatured withDiethylPhthalateonly)
Bath Preparations
BathOils, Tablets, and Salts (143) 1 2%
OtherBath Preparations (196) 52%
Eye Makeup Preparations
Eye Makeup Remover (100)
Mascara(195) 4 5—10%
OtherEye Makeup Preparations (152) 5%
FragrancePreparatons
Colognes and ToiletWaters (684) 104 86%
Perfumes (235) 32 78—88%
OtherFragrance Preparations (173) 10 76—81%
Non-coloring Hair Preparations
HairConditioners (651) 1 10%
HairSprays (aerosol fixatives) (275) 5
Rinses (non-coloring) (42)
Shampoos (non-coloring) (884) 1 0.3%
Tonics, Dressings, and OtherHair Grooming Aids (598) 12 0.3%
OtherHairPreparations (277) 6 74%
Hair Coloring Preparations
HairDyes and Colors (1690) 58
Makeup Preparations
Foundations (324) 1 0.003%
OtherMakeup Preparations (201) 28%
Personal Cleanliness Products
Bath Soaps and Detergents (421) 0.0002%
—
Deodorants (underarm) (247) 18 0.001—87%
OtherPersonal CleanlinessProducts (308) 1 0.0005—65%
Shaving Preparations
Aftershave Lotion (231) 35 88%
Preshave Lotions (all types) (14)
Other Shaving Preparation Products (63) 3 72%
Skin Care Preparations
Skin Cleansing (cold creams, lotions, liquids, andpads) (775) 4 8%
Face andNeck skin care (exci. shaving) (310) 12 0.05—12%
Body and Hand skincare (exci. shaving) (840) 4 5%
FootPowders and Sprays (35) 73%
—
Moisturizers (905) 3
Night skin care (200)
Paste Masks (mud packs) (271) 2
SkinFresheners (184) 4 0.006%
Other Skin CarePreparations (725) 8 58%