`
`MILDRED S. CHRISTIAN
`
`

`

`Journal of
`the American College
`of Toxicology
`
`Editor-in-Chief: Mildred S. Christian
`
`Assistant to the Editor: Ca ro lyn P. Nettles
`
`Argus Resea rch Laborato ries, In c.
`935 H o rsham Road
`H orsham, PA 79044
`(2 15) 443-8710
`
`Arth ur Furst
`Institute of Chemica l Bio logy
`University of San Fran cisco
`Sa n Francisco, CA 94 7 7 7
`(415) 666-64 15
`
`GENERAL TOXI CO LOGY
`Section Editor
`Bruce K. Bernard
`Washingto n, DC
`
`Secl io n Associate Edit o rs
`Joseph F. Bo rzelleca
`Wa yne M . G a lbraith
`Do nald Hughes
`Rud olph J. Jaeger
`Ca rol A. M apes
`Ian C. Munro
`
`ACUTE AND SU BCHRONI C TOXICITY
`Section Ed ito r
`Ric ha rd A. Pare nt
`Baton Rouge, LA
`
`SAFETY EVALUATION AND RISK ASSESSMENT
`Secli o n Edit o r
`Robert G . Ta rd iff
`Springfield, VA
`
`CARC INOG ENESIS
`Section Edito r
`Yin-tak Woo
`Wa shington, DC
`
`Section Associate Editors
`F. Peter Cuengerich
`Charles C. Irving
`David Y. Lai
`John H . Weisbu rger
`Hanspeter R. Wit schi
`
`IMMUN OTOX ICOLOGY
`Section Editor
`Stewa rt Wong
`Waverly, PA
`DEVELOPMENTAL AND REPRODUCTIVE TOXICOLOGY
`Section Editor
`E. Marshall Joh nso n
`Phi ladelph ia, PA
`
`Associate Editors
`Robert M. Diener
`Ciba-Geigy Pharma ceuticals Division
`Summit, NJ 0790 1
`(201) 277-5430
`
`Editorial Board
`
`Section Associate Edilo rs
`Tho ma s H. Shepard
`Rober! E. Slaples
`Pe1er Voy1ek
`Ernest F. Zimmerman
`GENETIC TOXICOLOGY
`Seclion Edilo r
`W.Gary Flamm
`Rockville, MD
`Section Associate Editors
`Virginia Dunkel
`George Hoffmann
`Verne A. Ray
`EPIDEMIOLOGY AND CLINICAL TOXICOLOGY
`Section Edilor
`Edward J. Calabrese
`Amhe rst , MA
`MECHANISMS OF TOXI CITY
`Section Editor
`Ronald W. Ha rt
`Je ((e rson, AR
`Section Associate Ed itor
`Gerald N. Wogan
`DEVELOPMENT OF NON-A NIMAL TESTING TECHNIQUES
`Sect ion Editor
`Alan M . Goldbe rg
`Ba ltimore, MO
`Sectio n Associate Editors
`Thoma s E. Hickey
`De nnis Stark
`QUALITY ASSURANCE
`Section Editor
`Jane E. Goeke
`Horsham , PA
`VETERINARY TOXICOLOGY
`Section Editor
`E. Muri Bai ley, Jr .
`EDITOR FOR BOOK REVIEWS
`Wayne M. Galbrai th
`Arlingto n, VA
`
`

`

`\
`'
`
`GENERAL INFORMATION
`
`The Journal of the American College of Toxicolog y publishes fully refereed papers covering the
`entire field of toxicology. These include research in risk assessment, general toxicology, carcino(cid:173)
`genicity, safety evaluation, reproductive and genetic toxicology, epidemiology and clinical toxi(cid:173)
`cology, mechanisms of toxicity, new approaches to toxicological testing, and alternatives to ani(cid:173)
`mal testing. Reviews and major symposia in the field are included .
`
`Journal of the American College of Toxicology (ISSN:0730-09 13) is published bimonth ly for $130 per
`year by Mary Ann Liebert, Inc., 1651 Third Ave nue, New York NY 10128 . (2 12) 289-2300. Second(cid:173)
`class postage paid at New York, NY and at additional mai ling offices.
`
`Postmaster: Send address changes to Journal of the American College of Toxicology c/o Subscrip(cid:173)
`tion Department, Mary Ann Lieb~rt, Inc., Publi shers, 1651 Th ird Ave nue, New York, NY 10128.
`
`Subscriptions should be addressed to the Publisher and are paya ble in adva nce. Rates are $130 per
`volume of 6 issues in the United States and Possessions, $163 ai r mail , elsewhere.
`
`Reprints, except ·special orders of 100 or more, are ava ilable fro m the authors.
`
`Business communications should be addressed to the Publisher.
`
`Advertising inquiries should be add ressed to Ma ry Ann Liebert, Inc. , 1651 Third Avenue, New
`York, NY 10128. (2 12) 289-2300.
`
`Manuscripts should be directed to the Editor: Mildred Christian , A rgus Resea rch, 935 Horsham Rd.,
`Horsham , PA 19044.
`.
`
`All authored papers and editori al news and comments, opinions, findings, concl usions, or recom(cid:173)
`mendations in the Journal of the American College of Toxicology are those of the author(s), and do
`not necessarily refl ect th e views of the journal and its publisher, nor does their publication in the
`Jo urnal of the American College of Toxicology infer any endorsement.
`
`Copyright © 1988 by The American College ofToxicology. Printed in th e United States of America.
`
`Thi s Journal is indexed in Current Contents and Science Citation Index.
`
`16~1 n.1u h o-. I!... To,~, J, Y,
`
`IOU!
`
`...
`
`!6~iTl,fr·A ....... . - 1-,\, J./
`
`I Ona
`
`

`

`JOURNAL OF THE AMERICAN COLLEGE O F TOXICOLOGY
`Volume 7, Number 3, 1988
`M•ry Ann Liebert, Inc., Publishers
`
`3
`
`Final Report on the Safety
`Assessment of Polyquaternium-1 O
`
`Polyquaternium-10 is a polymeric quaternary ammonium derivative of hy(cid:173)
`droxyethyl cellulose that is used in cosmetics as a conditioner, thickener and
`emollient at concentrations of :s; 0.1%-5%. Polyquaternium-10 has, at .'.nost,
`only a low potential to penetrate the stratum corneum but is adsorbed by ke(cid:173)
`ratinous surfaces. The oral LDso of Polyquaternium-10 was not obtained at 16
`g/kg in rats. Inhalation, dermal, and ocular animal test data indicated, at
`most, only a low degree of toxicity at test concentrations of Polyquaternium-
`10 greater than that used in cosmetic products. Polyquaternium-10 with and
`without metabolic activation was not a mutagen in three separate assay sys(cid:173)
`tems. Polyquaternium-10 was neither an irritant nor a human sensitizer when
`tested at 2.0 %. Cosmetic products containing .up to 1 % Polyquaternium-10
`were not human irritants, sensitizers, or photosensitizers. On the basis of the
`information presented, it is concluded that Polyquaternium-10 is safe as a
`cosmetic ingredient in the present practices of use.
`
`INTRODUCTION
`
`Polyquaternium-1 O is a cationic form of hyd~oxyethyl cellulos~ that adsorb_s
`
`and sorbs well to proteinaceous surfaces. It 1s used 1n cosmetics as a condi(cid:173)
`tioner, thickener, and emollient in hair care products, lotions, and makeup.
`
`CHEMISTRY
`
`Definition
`Polyquaternium-10, also known as Quaternium-19, i~ a pol_ymeric quater(cid:173)
`nary ammonium salt of hydroxyethyl cellulose reacted w ith a tnm~thyl am~o(cid:173)
`nium substituted epoxide. There are va rious grad~s of Polyquaternium-10 with
`different average molecular weights generally ranging from 250,000 to 600,000.
`Polyquaternium-10 has three CAS numbers: 53568-66-4, 54351-50-7, and
`55353-19-0. (J-3)
`
`Chemical and Physical Properties
`Po lyquaternium-1 O is a w hite granular powder with a characteristic ~mi ne
`odor. It is soluble in water and insol uble in alcohol and nonpolar orga nic sol-
`
`335
`
`

`

`336
`
`COSMETIC INGREDIE NT REVIEW
`
`vents. Polyquaternium-1 O used in cosm etics has 0. 5% m ax im um water insolu(cid:173)
`bles 1.7 to 2.2% nitrogen-containing components, 2% maxim um ash (NaCl)
`and ' 6% maximum volatile material. T he particle size speci fication s are 95°!;
`minimum throu gh a 20 mesh filter and 85 % minimum through a 40 mesh filter.
`Th e viscosity of a 2% aqueous solution (25° ( ) is between 60 and 150 centi(cid:173)
`poises. t•>
`Polyquaternium-1 O alters the su rface tensio n of aqu eo u s so lution s of ani(cid:173)
`onic surfactants. Addition of 1 % and 2% Po lyqua ternium-10 lowered the sur(cid:173)
`face tension of aqueous sol utions of sodi um lauryl sulfate, sodium tridecylben(cid:173)
`zenesulfonate, and potassium laurate. t5
`>
`
`I i
`
`1
`I
`
`Reactivity
`
`Polyquaterniu m-1 O is a cationic, surface-active polymer that is adsorbed by
`keratinous surfaces, such as hair and skin (stratum corneum). The ad sorption of
`the polymer was not readily affected by pH in the range o f 4 to 10. It undergoes
`slow hydrolyt ic cleavage outside this pH range. Sorption of Polyquaternium-10
`to keratinou s surfaces was decreased by the addition of electrolytes (salts), such
`as aluminum, iron, calcium , or sodium . Polyquaternium -10 i s biologically de(cid:173)
`gradable. The presence of ethyl alcohol o r propylene glycol add s to th e stability
`of Polyquaternium-10. (6- 9 1
`
`Analytical M ethods
`
`The most common anal yti ca l method for qu aternary ammonium com(cid:173)
`pounds is colorimetri c testing following separation by acid extraction . tio,
`
`M ethod of Manu facture
`
`Polyquaternium-10 is generally produced by reac tin g hyd ro xyethylcellulose
`with epichlorhydrin, followed by quaterni za tion usi ng t rime th ylamine. It is
`stable within a pH rane of 4 to 8.
`
`Impurities
`
`Inorganic impurities of Polyquaternium-1 O used in co smetics include water
`(up to 0.5%), nitrogen (1.7 to 2.2%), and ash (NaCl up to 2%) . Information was
`not available on organic impurities of Polyqu atern ium-1 O. <•> Epic hlorhydrin was
`not detected in any of six different Polyquaternium polymers analyzed with an
`aver~ge detection limit of about 0.5 ppm .t 11 > A maximum of 10.8 ppm Trimeth(cid:173)
`ylami ne was detected in lots of Polyquaternium-1 o th at were produced and sold
`during 1985.t 12 >
`
`USE
`Purpose in Cosmetics
`
`_Polyquate~nium-10 is used as a conditio ner, emollient and viscosity con(cid:173)
`trolling agent 1n cosmetics. <J. 1J.1 • 1
`
`

`

`ASSESSMENT: POL YQUA TERNI UM-10
`
`337
`
`Scope and Extent of Use in Cosmetics
`
`Polyquaternium- 10 is used primarily in hair care products, skin cleansers,
`and ski n moisturi zers in concentrations of :s;0.1 %-5%. One hundred thirty-nine
`(139) of the vo luntarily filed cosmetic product formulations were reported to the
`Food and Drug Adm inistration to contain Polyquaternium-10Y 5 > The cosmetic
`product formulation data that are made available by the FDA are compiled
`through voluntary fi ling of such data in accordance with Title 21 pa rt 720.4 of
`the Code of Federal Regulations. 116> Ingredients are listed in prescribed concen(cid:173)
`tration ran ges under specific product type categories. Since certain cosmetic in(cid:173)
`gredients are supplied by the manufacturer at less than 100% concentration, the
`value reported by the cosmetic formulator may not necessarily reflect the actual
`concentration found in the finished product; the actual concentration would be
`a fraction of that reported to the FDA . Data submitted within the framework of
`preset concentration ran ges provide the opportunity for overestimation of the
`actual concent ratio n of an ingredient in a particular product. An entry at the
`lowest end of a concen tration range is considered the same as one entered at
`the highest end of that range, thus int roduci ng the possibility of a 2- to 10-fold
`error in the assumed ingredient concentration (Table 1 ).
`.
`Polyquaternium-1 O has been generally or individually approved for use in
`cosmetic formulations marketed in Japan. <11
`>
`
`Surfaces, Frequency, and Duration of Application
`
`The hair ca re products con taining Polyquaternium-10 are, for t~e most _pa.rt,
`applied for a few minutes, then rin sed off. However, P~lyquatern,um-1 0 ,s in(cid:173)
`corporated in these products for its "substantivity" (sorpt,on) and w_ould be ex(cid:173)
`pected to rem ain in contact with the hair between treatments. Skin cl~ansers
`and moisturi zers have the potential to remain in continuous contact with the
`skin and nails for extended periods of time.
`
`BIOLOGY
`
`J
`I
`l
`
`Penetratio n, Sorption, and Permeation Through Stratum Corn eum
`I f
`of 14( -Polyquaterni um-10
`Approximately 1 ml of a 5% (w/~) water. so u ,on
`lied to the backs of 12
`(uniformly labeled on the pendant side chain) was app
`h
`t
`·od 1
`.
`f
`1/k
`occluded for t e tes pen
`•
`Fischer 344 rats. The total dose o _4.0 m g was
`boli sm cages designed
`3, and 24 h. Six rats were plac~d in Roth-type gla~s ~:t~ for the entire 24-h ex(cid:173)
`for the separate collection of urine, feces, an.d expire
`k.ll d 3 at 1 h and 3 at
`posure period. The remaining 6 exposed ant~ ~\ 1~;~h~ i~t~rim sacrifices and
`6-h postexposure. Blood measured at 1 h an
`no radioactivity above back-
`for the 6 material balance rats killed at 24 h had
`ground. There was no radioactivity above backgrou\ ,n i 02 01, of the adminis-
`d .
`ny of 9 tissues exam-
`ined in the 6 animals _that wer~ killed at 2\ \ ~e~~,: i~~h~ fec;s; no radioactiv(cid:173)
`tered dose was fo u~d in the urine and l~ss t fa he ·total dose that was recovered
`1
`tty was recovered tn CO2. Of the 83. 7 to 0
`terials and 7 .1 % was fou nd
`.
`1
`from the male rats, 75 .2% was from the occ us,ve ma
`
`

`

`TABLE 1. Produce Formulacion Daca for Polyquacernium- 10""
`
`Product category
`
`Bubble baths
`Mascara
`Hair cond icioners
`Hair sprays (aerosol fixacives)
`Permanenc waves
`Hair shampoos (noncoloring)
`Tonics, dressings, and ocher hair grooming a,ds
`Wave sels
`Ocher hair preparacions (noncoloring)
`Hair shampoos (coloring)
`Makeup bases
`Bath soaps and detergents
`Skin cleansing preparacions (cold creams, loc,ons,
`liquids, and pads)
`Face, body, and hand skin care prepara c,ons (excluding
`shaving preparations)
`Moisturizing skin care prepara1 ions
`Night skin care preparations
`Paste masks (mud packs)
`Skin fresheners
`Wrinkle smoothers (removers)
`Other skin ca re preparacions
`
`1981 TOTALS
`
`Total no. of
`fo rmulat ions
`,n catego ry
`
`Total no.
`conta,nmg
`,ngredient
`
`N o. of produc t formula tions within each
`co ncentration range ( % )
`
`U nreported
`concentration
`
`> 1- 5
`
`>0. 1-1
`
`:SO. /
`
`475
`397
`478
`265
`474
`909
`290
`180
`177
`16
`831
`148
`680
`
`832
`
`747
`219
`171
`260
`38
`349
`
`I
`5
`16
`l
`6
`62
`4
`6
`I
`3
`2
`l
`6
`
`8
`
`12
`I
`l
`l
`l
`I
`
`139
`
`-
`-
`-
`-
`-
`-
`-
`-
`-
`-
`
`2
`
`-
`
`I
`-
`-
`-
`-
`-
`3
`
`-
`6
`-
`
`l
`9
`-
`2
`-
`-
`-
`-
`-
`
`-
`-
`-
`-
`-
`-
`
`-
`9
`l
`5
`46
`3
`3
`1
`3
`-
`
`I
`
`3
`
`3
`-
`-
`-
`-
`1
`
`18
`
`81
`
`5
`
`7
`
`2
`
`3
`
`5
`
`a
`I
`I
`I
`l
`-
`37
`
`.....
`...,
`
`00
`
`(")
`0
`VI
`~
`m
`-4
`;:;
`z
`C)
`;IQ
`m
`0
`m
`z
`-4
`;c m
`< ;;;
`:£
`
`- --A----
`
`

`

`ASSESSMENT: POLYQUATERNIUM-10
`
`339
`
`i~ or around the ski_n at the dose site. The fraction of the applied material that
`~1~ penetr?te th~ skin amounted to less than 1.5% of the administered radioac(cid:173)
`t1v1ty. The _investigators noted the low total recovery of the applied radioactivity;
`however, 1f the data were normalized to 100%, less than 1.75% of the applied
`?ose w~uld h~ve penetrated the skin. The authors concluded that Polyquater-
`1um-l0 1s unlikely to penetrate the human skin in toxicologically significant
`amounts.< 18
`9 >
`• 1
`. Th_e anti-irritant e~ect of hydroxyl and quaternary am monium compounds,
`including Polyquatern,um-10, has been investigated in recent years. It has been
`hypothesized that the sorption properties of Polyquaternium-10 enable the
`polymer to block keratin reactive sites, thus reducing the topical and eye irritant
`effects of surfactants in shampoo and cleanser formulations. <20-2 1>
`Several studies have investigated the passage of Polyquaternium-10 and sur(cid:173)
`factants, such as sodium lauryl sulfate, into and through isolated stratum cor(cid:173)
`neum from the skin of fetal pigs, neonatal rats, and adult humans. The fetal pig(cid:173)
`skin was frozen, and on warming, the stratum corneum was gently separated
`from the underlyi ng epidermis. Neonatal rat skin was removed following death
`by CO2 inhalation. The skin was placed in a desiccator jar and exposed to am(cid:173)
`monia vapor for 1- 3 h, then put in water to remove the epiderm is from the der(cid:173)
`mis. After 1 h the epidermis was recovered on a paper towel, and the mal(cid:173)
`pighian layer was scraped off, leaving the stratum corneum to dry. The various
`preparations were exposed to ••e-Polyquaternium-10 and/or surfactant (i n aque(cid:173)
`ous solution) in via ls or permeability cellsP 2
`> to quantify sorption and perme(cid:173)
`ability and to evaluate the effects of the polymer on the preparations. The results
`indicated that Polyquaternium-10 slowly diffused into the outer layer of the
`stratum corneum (all three species) rather than forming multilayers on the sur(cid:173)
`face. Although ionic surfacta nts did reduce polymer sorption, pretreatment of
`the stratum corneum with 1 % Polyquaternium-10 greatly reduced the amount
`of surfactant that passed through the preparation. This reductio_n in per~.e~tion
`of surfacta nt supported the hypothesis that Polyquaternium-10 1s an ant1-1mtant
`in the presence of ionic (i.e., irritant) surfactants. The kinetics of polym_er/s~rfac(cid:173)
`tant sorption and permeation indicated that the polymer did not ~ct ~nmarily as
`a barrier to penetration, but rather it helped to maintain the physical integrity of
`the stratum corneum preparation .
`.
`.
`Pretreatment of the stratum corneum of neonatal rats with Polyquat_ernium-
`10 slightly increased the hydration of the membrane and reduced swelling after
`surfactant exposure. 12
`2•>
`9
`23
`
`•
`
`•
`
`•
`
`ANIMAL TOXICOLOGY
`Oral Toxicity
`Three lots of Polyquaternium-10 were tested for acute oral toxicity by int~(cid:173)
`bation in rats. Three studies at maximum doses of 16 g/kg, 13.1 g/kg, 16 g/kg did
`not
`h ·
`th LO
`<25
`21 >
`ac ,eve e
`h
`00 and a con-
`0 .
`•
`50·
`.
`The acute oral toxicity of 1.0% Polyquatern1um-l
`1n a s amp.
`ditioner was tested by gavage. The LD5o of the two formulations was not
`reached at s g/kg. 12s.29 > Results are summarized in Table 2.
`
`

`

`f
`i
`
`1
`
`340
`
`COSM ETIC INGREDIENT REVIEW
`
`TABLE 2. Acu1e Oral Toxicily
`
`Compound
`
`No. of rats
`
`Vehicle
`
`Polyquaternium-1 0
`(lot 1J
`
`10
`
`Corn oil
`
`LO,.
`
`> 16 g/kg
`
`Polyquaternium-10
`(Loi 1l
`
`Polyqua1ernium· l 0
`(lot 2)
`
`Polyquaiernium-10
`(Loi 2)
`
`Polyquaiernrum-10
`(Loi 2)
`
`Polyquaternrum-10
`(Loi 3)
`
`3
`
`10
`
`10
`
`5
`
`5
`
`Water
`
`ot reached
`
`Corn orl
`
`13. 1 g/kg
`
`Corn oil
`
`'Jot reached
`
`Waler
`
`'sot reached
`
`Corn orl
`
`> 16.0 g/kg
`
`Comments
`
`Reference
`
`3/10 died; sluggish. wet
`fur and d iarrhea on
`one day; low order of
`1ox1city
`
`one dred: sluggish a1
`4 g/kg
`
`7/10 dred; sluggish. we1
`fur nose CO\ ered
`w11h blood on 1 day
`
`-.:one dred al 8 g/kg:
`appeared normal
`
`'Jone died at 2 g/kg
`appeared normal
`
`-.: one dred al 16.0 g/kg:
`" et tur and diarrhea
`al l day
`
`25
`
`25
`
`26
`
`26
`
`26
`
`r
`
`3
`
`\Valer
`
`'-one dred a1 4 g.kg
`Polyquaiernrum-10
`animal~ ~luggrsh
`(lot 3)
`----------------------------------------------------------------------------------------
`> 5 g/kg
`one dred· nontoxic
`28
`Polyquaternrum-10
`10
`Formula11on
`l °' .. in shampoo
`
`-..: 01 reached
`
`27
`
`Polyquaternrum-10
`1°' .. in condi-
`Ironer
`
`10
`
`Formula11on
`
`> 5 g/kg
`
`, one dred nontoxic
`
`29
`
`Inhalatio n Toxicity
`
`Inhalation tests of three lots of Polyquaternium-1 O were conducted using in(cid:173)
`dividual groups of 6 rats and an exposure period of 8 h. The rats were exposed
`to a substantial aerosol concentration that was p repared by placing 50 g of test
`material on a 200 cm 2 tray for 16 h before testing in a sealed chamber and dur·
`ing the 8-h exposure period . A fan was operated intermittently to agitate the in·
`ternal chamber atmosphere. o signs of toxicity were observed during the ex(cid:173)
`posure period. 1251
`
`Dermal Toxicity and Irritatio n
`Three lot samples of Polyquaternium-1 o were tested individually using thr~
`groups of 5 rabbits each. o deaths occurred when 4.0 g/kg was applied di(cid:173)
`rectly to the skin and the application site covered . Erytheme developed, but no
`~ther remarkable gross lesions were observed. The th ree lot samples were con·
`s1dered, at most, slightly toxic. 12s-21 1
`.
`Three lots of Polyquaternium-10 were tested o n the clipped uncovered in·
`tact skin of the abdomen of 5 rabbits at individ ual lot concentrations of 2%, S%,
`
`

`

`ASSESSMENT: POLYQUATERNIUM-10
`
`341
`
`a~d 10% (in water). At 2% concentration, no irritation was observed in 4 rab(cid:173)
`~1ts,_ and moderate ery~hema wa_s seen in 1 rabbit. At 5.0% concentration, no ir(cid:173)
`n~at1on was observed in 3 rabbits, and moderate erythema was seen in 2 rab(cid:173)
`bits. At 10% concen tration , no irritation was observed in 2 rabbits, and moder(cid:173)
`ate erythema was observed in 3 rabbits. The investigator termed the observed ir(cid:173)
`ritation a trace reaction . <2 5 - 27 1
`A shampoo con taining 0.5% Polyquaternium-10 had a primary irritation in(cid:173)
`dex (PII) of 5.37 (max . 8) in a study performed according to CFR Title 16:
`l 500.3(c)(4) and 1500.41. <301 The test used 12 rabbits, dosed at 0.5 ml of the
`shampoo, w ith 6 having intact ski n and 6 having abraded ski n. The shampoo
`was a severe skin irritant .<18 · 3 '
`'
`The dermal toxicity of a shampoo containing 0.5% Polyq uaternium-10 was
`evaluated according to the procedures in the CFR Title 16:1500.3(c)(l)(ii)(c) and
`1500.40. po, The product was tested on the clipped skin of 5 rabbits and 5 that
`were both clipped and abraded. The dermal LD50 of the formulation was greater
`than 2 g/kg. <28
`3
`' 1
`•
`A 21-day su bc hronic dermal toxicity study was conducted with a condi(cid:173)
`tioner contai ning 1 % Polyq uaternium-10. Five male and five female rabbits
`were given 0.5 m l undiluted product once a day for 21 consecutive days. The
`test material was ap plied to the clipped dorsal trunk of the rabbits, and the skin
`of 5 of 10 ani mal s was abraded before application of the test material. The test
`material rem ained in contact with the ski n 6 h daily, and sites were uncovered .
`A control group of 5 male and 5 female rabbits had tap water applied to intact
`and abraded skin. o rabbits died during the test period, both groups had nor(cid:173)
`mal weight gai ns, and no lesions were observed at necropsy. Initial and termin al
`hematological and urinalysis val ues were within normal range. The product was
`21
`not a cumulative dermal toxin. P
`Six ew Zealand rabbits were used to evaluate the primary skin irritation of
`a conditioner contai ning 1 % Pol yquaternium -10. One-half (~.5) millili~er of the
`test material was administered to 1 intact and 1 abraded test site per animal. The
`clipped test sites were covered by occlusive patches for 24 h, and test sites were
`sco red for irritation immediately after patch removal and 48 h later. The group
`PII was 0.0 (max . 8). The product was not a primary skin irritant< 291 (Table 3).
`
`Ocular Toxicity
`
`Three lots of Polyq uaternium-1 O were tested for ocular _toxicity, b_ot~ a_s a
`dry powder and as an aqueous solution using groups of 5 animals. o 1mtat1on
`was produced by the powder form of any lot. When applied as the aqueous
`solution, no irritation was produced by lot 1 at a 20% co nc~ntrat1on or ~Y lot 2
`at a 5% concentration. A trace of irritation was produced by lot 3 at a 10 Yo con-
`centration. <25- 27 >
`•
`•
`A shampoo conta ini ng 0.5% Polyquaternium-10 was tested for o~ular irrita-
`tion in two groups of 12 rabbits according to the pro~edures given in t~e ~FR
`Title 16: 1500.3(c)(4) and 1500.42.<3°'. One gr_ou~ received / ull~str~ngth instilla(cid:173)
`tions of the shampoo; a second received instillation of a 5 Yo. dil~tio_n. The e_y~s
`of half of the rabbits of each group were rinsed after product 1nstdlat1on . No 1m-
`
`

`

`..,
`~ ...,
`
`TABLE 3 . Dermal Toxicil y and lrrilalion
`
`Compound
`--
`Polyqualernium-10
`(LOI I )
`
`Polyq ualernium-10
`(LOI 2)
`
`Polyqualernium- 10
`(Loi 3)
`
`Polyq ualernium-10
`(Loi I)
`
`Polyquaternium-10
`(Lot 2)
`
`No. of rabb,rs
`
`Vehicle
`
`Concentration
`
`Method
`
`Results
`
`Reference
`
`5
`
`5
`
`5
`
`5
`
`5
`
`None
`
`4 g/kg
`
`Applied direclly 10 clipped inlacl
`skin, occluded for 24 h
`
`None
`
`4 g/kg
`
`Applied clireclly 10 clipped in1ac1
`skin, occluded for 24 h
`
`None
`
`4 g/kg
`
`Applied direc1 ly 10 clipped in1ac1
`sk,n, occluded for 24 h
`
`Waler
`
`2%
`
`Water
`
`5%
`
`0.01 ml applied 10 clipped in1ac1
`bellies for 24 h
`
`0 .01 ml applied to clopped 1n1act
`bellies for 24 h
`
`No deal hs; erylhema al 24 h;
`no remarkable gross palho-
`logical resuhs
`No deal hs; ery1hema al 24 h;
`no remarkable gross palho-
`logical resuhs
`No dealhs; erythema al 24 h;
`no remarkable gross palho-
`logical resu lls
`No orrilalion in 4; moderate
`capillary injection in 1
`rabbit
`
`No irritation in 3; moderate
`capillary injection in 2
`rabbits
`
`25
`
`26
`
`27
`
`25
`
`26
`
`Polyquaternium-10
`(l ot 3)
`
`5
`
`Water
`
`No irrilation in 2; moderate
`capilla ry injection in 3
`rabbits
`-- ------------------------------------------------------------------------------------------------------------
`--- ---
`Shampoo contain-
`12
`Dorec1
`1.0%
`( FR I 6: 1500.3(( )(4), I 500.4 I ;
`PII 5. 37 (max. 8); shampoo
`28, 3 I
`ing 1 .0% Poly-
`6 with intact and 6 with
`was classified as severe irri-
`q uaterniu m- 10
`abraded skin
`tant
`
`10% (suspen-
`soon)
`
`0.01 ml applied to clipped intact
`bellies for 24 h
`
`27
`
`Conditioner con-
`taining 1.0%
`Polyqua-
`iernium-10
`
`6
`
`Direct
`
`1.0%
`
`0.5 ml applied to each intact
`and abraded area; occluded
`for 24 h
`
`PII 0.0 (max. 8); conditioner
`was not skin irritant
`
`29
`
`(")
`0
`V>
`~
`m
`::::!
`(")
`z
`C'l
`,0
`m
`
`51 m z
`
`-f
`,0
`m
`!:
`i
`
`___ _
`
`.J
`
`

`

`ASSESSMENT: POLYQUATERNIUM-10
`
`343
`
`th h
`talion was observed in the eyes of any animal The full- t
`not an eye irritant. <3 • . JJ>
`s reng
`s ampoo was
`•
`t
`The ocular irritation or a conditioner containing 1 o, Pol
`·
`z I
`.
`t d · 9 N
`yqua ern,um-lOwas
`I
`10
`eva ~a e_ in . ew ea and rabbits. One-tenth (0.1) milliliter of the test material
`was instilled into one eye of each rabbit and the other
`d
`.
`,
`eye serve as an un-
`I
`d
`treate contra . The eyes of 3 rabbits were rinsed 15 sec afte
`t·
`1·
`f h
`I
`r app ,ca 10n o t e
`·
`· .
`.
`test so ut,on. No 1rrr~at1~n was observed in rinsed or unrinsed eyes 1, 2, 3, 4,
`and ~9~ays after appl1cat1on of the product. The conditioner was not an eye irri-
`tant.
`A summary of ocular test data is given in Table 4.
`
`MUTAGENICITY AND GENOTOXICITY
`
`Polyquaternium-10 was tested in triplicate and at five concentrations for
`mutagenic activity in the Salmonella typhimurium assay (Ames test) on TA-98,
`TA-100, TA-1535, TA-1537, and TA-1538 tester strains without and with meta(cid:173)
`bolic activation. The metabolic activation was induced with 59 liver homoge(cid:173)
`nate from male Sprague-Dawley rats pretreated with Aroclor 1254. In this
`setting, Polyquaternium-10 was not mutagenic in any of the five bacterial strains
`tested wit h and without metabolic activation. 1341
`Chi nese hamster ovary (CH O) cells were exposed for 5 h to five concentra(cid:173)
`tions of Polyquaternium-10 (0. 12-0.285% in DMSO) without and with addition
`of S9 liver homogenate for metabolic activation (as in the Ames assay). Although
`some degree of mu tagenicity was observed in both of the duplicate assays in the
`absence as well as in the presence of 59 homogenate, the response was not
`dose dependent. Polyquaternium-10 was thus considered to be nonmutagenic
`in the C HO test. In sister chromatid exchange (SCE) assays, Polyquaternium-10
`in CHO cells both with and without enzyme induction by 59 liver homogenate
`did not increase the frequency of SCE over the range of concentrations tested
`(0. 14-0.23%). ( J 4 )
`Polyquaternium-1 O was evaluated also for its genotoxic effect in the rat he(cid:173)
`patocyte primary culture/ DNA repair test in six concentrations between 0.~0~9
`and 0.23%. The results from duplicate samples indicate significant genotox rc~ty
`of this cosmetic ingredie nt at three of six doses. However, the measured activ(cid:173)
`ity, which was not dose dependent, was determined with hepatocytes from a
`si ngle preparation and quantitated by liquid scintillation counting rather ~han by
`the more reproducible quantitative autoradiography. 1341 The result~ of th,s_study
`do not allow a va lid conclusion about the activity of Polyquatern, um-10 rn he-
`patocyte DNA repair test.
`.
`. .
`.
`Polyquaternium-1 o was tested for its in vivo clastogenrc act1v1 ty rn f~male
`Swiss mice by injecting single doses of 0.125, 0.25, and 0.4 g/kg, respectively,
`into the peritoneal cavity of 5 males and 5 females. Blood samples were col(cid:173)
`lected 24, 48, and 72 h after the injection. Blood smears_ were prepared a~d
`stained for evaluating the possible presence of micro~~cler. No stat,st,cally sig(cid:173)
`nificant increases in micronucleated polychromatophrlrc eryth rocytes were re(cid:173)
`ported. 134 1
`
`

`

`TABLE 4. Ocular Toxicity
`
`Compound
`
`N o. o f rabbit s
`
`Vehicle
`
`Concentrat,on
`
`M ethod
`
`Results
`
`Reference
`
`Powder
`
`Direct
`
`Single instillation into conju nct,val sac
`
`No irrila tion
`
`5
`
`5
`
`5
`
`5
`
`5
`
`5
`
`Water
`
`2%
`
`Powder
`
`Direct
`
`W.iter
`
`5%
`
`Powder
`
`D irect
`
`Water
`
`10% (suspen(cid:173)
`; ,o n)
`
`0 .5 ml si ngle instill;ition into conjunc(cid:173)
`tiva! sac
`0 .5 ml single instill;, tion into conjunc(cid:173)
`tival sac
`0.5 ml single instillat,on ,nto con1unc(cid:173)
`t1val sac
`0 .5 ml single instillation into conjunc(cid:173)
`ltval sac
`0 .5 ml single 1nst1lla11on 11110 r onjunc(cid:173)
`tiva l sac
`
`Polyquaternium- 10
`(Lot 1)
`
`Polyquatern ium- 10
`(Lot I)
`Polyquaternium- 10
`(Lot 2)
`
`Polyquaternium- 10
`(Lot 2)
`
`Polyquaternium- 10
`(Lot 3)
`
`Polyquaternium- 10
`(Lot 3)
`
`Shampoo contain(cid:173)
`ing 0.5% Poly(cid:173)
`quaternium -10
`
`Conditioner con(cid:173)
`taining 1.0%
`Polyqua(cid:173)
`ternium-10
`
`12
`
`Direct and 5%
`dilutio n
`
`1.0 and
`0 .05%
`
`Cl R 16: I 500 .3(()(4) and I 500 .42; ha lf
`o f test eyes w ere rinsed
`
`9
`
`D irect
`
`0. 1 ml of
`conditioner
`
`Direct applicatio n, 3 eyes rinsed
`
`25
`
`25
`
`26
`
`26
`
`27
`
`27
`
`----- -·
`28, 3 1
`
`29
`
`No irritation
`
`N o irritation
`
`No orrotation
`
`No irritation
`
`Trace irritation
`in I eye,
`no ne in 4
`
`No irritation at
`either con-
`centration,
`rinsed or
`unronsed
`
`N o irritat ion,
`110 1 an eye
`irritant
`
`...,
`t
`
`("')
`0
`V>
`?
`m
`- j
`ri
`
`C)
`
`"' m
`0
`m z
`"' m
`~
`m
`~
`
`-j
`
`

`

`ASSESSMENT: PO L YQUA TERNIUM-1 0
`
`345
`
`CLINICAL ASSESSMENT OF SAFETY
`Primary Skin Irritation, Sensitization, and Photosensitization
`
`. A lot o! Polyquater~ iu~-10 was tested for ~rimary skin irritation on 106 sub(cid:173)
`Jects. A 5 ~o w/v solutio n in water was applied to the skin under occlusive
`~ate~~~ for ~8. h .. No skin irritation was observed at 48 and 72 h after applica(cid:173)
`tion. No 1mtat1on was observed when a 5% Polyquaternium-1 o solution was
`appl ied daily under occlusive conditions for 21 days to 27 subjects.<36>
`Three separate lots of Polyquaternium-10 were tested by a repeated insult
`p~tch test (RIP_T) p ro~oc_ol us_ing 50 su bjects. A 2% w/v sol ution in water was ap(cid:173)
`plied to the skin on lintine disks under occlusive patches for 24 h. The treatment
`r~action si tes .were scored at 24 h. Following a 24-h nontreatment period, the
`sites were again exposed to the test samples. A 2-week nontreatment period fol(cid:173)
`lowed the 15th application. The test sites were evaluated and then treated in a
`similar m anner and scored at 24 and 48 h. No evidence of either irritation or
`sensitization was observed in any subject w hen exposed to the three lot samples
`- 39>
`of Polyquatern iu m-1 O. <37
`There was no irritation or sensitization when a 5% w/v solution of Polyqua(cid:173)
`ternium-10 was used in an RIPT on 203 subjects. <•0 >
`A shampoo containing 0 .5% Polyquaterni um-10 was evaluated for primary
`irritation and sensi tization by a prophetic patch test <••> and an RIPT.<•2
`> One
`hundred subjects participated in the prophetic patch test, and no reactions were
`observed to either of the two closed patches applied. Fifty-three subjects com(cid:173)
`pleted the RI PT, and no reaction s were observed during induction or at chal(cid:173)
`lenge. Photosensitization also was evaluated in these two studies by exposu re of
`the sites to UV light during the induction phase of the RJPT stud y and during the
`chal lenge phase of the prophetic patch test. The undiluted shampoo was not a
`primary ski n irritant, sensitizer, or photosensitizerY '·43>
`The primary irritation , sensiti zation, and photosensitization of an undi luted
`sham poo contai ning 0 .5% Polyq uaternium-10 was evaluated in an RIPT accord(cid:173)
`ing to the procedu res of Shelanski and Shelanski<•2> using a 25-member panel.
`The UV light was applied during the induction phase of the stud y. Th.e s~ampoo
`was tested at full strength. The product was nonirritating, nonsens1t1zing, and
`nonphotosensitizing. <44
`>
`•
`Forty-six panelists partici pated in an RIPT of a condition in? product c~~tain(cid:173)
`ing 1 % Pol yq uaternium-10. Patches containing 0.2 ml undiluted cond1t1oner
`were applied to the inner arm or back of each subject on Mondays, Wednes(cid:173)
`days, and Fridays for a total of 10 induction patches. After a 10-20 day nontreat(cid:173)
`ment period, two chal lenge patches were applied simultaneo~sly, one '.o. the
`origi nal test site and the other to an untreated site. One panelist had minim.al
`erythema (score of ± on a 0-4 scale) to induction patch 9, and another panelist
`had a ± reaction to induction patch 10. Two panelists reacted at challenge; 1