Cosmetics &amp Toiletries - May 2014 - Cosmeticsandtoiletries201405-13988677100005e00d10a77-Pp

May 2 0 1 4
Genetic Pathways
Into Skin p 38
Affecting Cholesterol, Enzymes in Skin p 22
Antimicrobial Stability During Use p 56
Skin Mechanics with Age p 66
Re-activating Sunscreens p 84
Magazine Tablet App Now Available! p 16, 17
Fo r sa lo n q ua lity ha ir c o lo r, yo u ne ed ea sy-to -use, re lia b le

p e rfo rm a nc e rig ht o ut of the p a c ka g e. NEW Chro m a p o l 5
Po lym e r im p a rts sup e rb rhe o lo g y c o ntro l a nd p ro d uc t
a e sthetic s to d e live r no d rip a nd sp ot-o n ta rg eted ha ir
c o lo ring , a nd a new c o nsum e r e xp e rie nc e.
New Chromapol 5 Polymer (INCI: Acrylates/Beheneth-25 Methacrylate Copolymer), a

novel polymer specially designed for hair color formulations. It is suited for permanent
as well as semi-permanent color systems, producing an elegant smooth flow as well
as a non-drip shear-thinning rheology and high clarity gels.
KEY BENEFITS
t
Enables formulation of hair colors in the form of clear gels or glossy creams with
very pleasant aesthetics.
Hair color formulations free from fatty alcohols maximizing formulation flexibility
while maintaining color efficiency
Potential to improve sustainability profile due to cold process* and substantial

savings in energy, batch time and amount of fatty phase required
Provides a better consumer experience as well as targeted hair coloring
Pleasing hair sensory and color vibrancy
*Dyes may need to be solubilized via side batch at 60 C
Hair color formulations free from fatty alcohols maximizing formulation flexibility
while maintaining color efficiency.
FIGURE 1: Comparison of ingredients present in hair color creams and gels with
Chromapol 5 Polymer versus traditional formulations.
REDUCED INGREDIENTS
Ingredients present
in traditional hair color creams
Ingredients present in hair color creams or

gels using Chromapol 5 Polymer
Aqua
Aqua
Propylene Glycol
Propylene Glycol
Dyes and anti-oxidants
Dyes and anti-oxidants
Fatty alcohols
Chromapol 5 Polymer
Oleic acid
Ammonia or MEA
Oleth-10
Oils/esters/silicones (color creams)
Oleth-5
Ceteareth-20
Steareth-20
Cetyl alcohol
Oils/esters/silicones
All tra d ema rks o w ned b y The Lub rizo l C orp ora tion. 2014 The Lub rizo l Corp ora tion.
FORMULATION FLEXIBILITY
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we ll a s sem i-p e rm a ne nt c o lo r syste m s,
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Rheology modifiers
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With Chromapol 5 Polymer fewer ingredients are required to make a hair color gel/
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Polymer
IMPROVED SUSTAINABILITY PROFILE

Potential to improve sustainability profile due to cold process* and substantial
savings in energy, batch time and amount of fatty phase required.
FIGURE 2: Potential production cost savings using Chromapol 5 Polymer technology
COST SAVINGS
Traditional hair color cream
Hair color with Chromapol 5 Polymer
Energy
(Heating and Cooling)
Energy No Heating and Cooling Required (with the

Exception of Dye Solubilization)
Formulation
(10-12% Fatty Alcohols
and Secondary Emulsifiers)
Formulation No Fatty Alcohols and/or Secondary

Emulsifiers
Longer Batch Time
Batch Time Reduction of 50% or More
*Dyes may need to be solubilized via side batch at 60 C
PLEASING HAIR SENSORY AND COLOR VIBRANCY

FIGURE 3: Color Vibrancy
30.0
Average L Values
25.0
20.0
15.0
10.0
5.0
0.0
Neat Chromapol 5
Polymer Gel without
Merquat Polymers
With Merquat
280 Polymer
With Merquat
2003PRPolymer
Comparison of L values of Hair Color Gel 5.7

Merquat polymers further enhance the wet combing properties and color vibrancy
of hair colors using Chromapol 5 Polymer compared to commercial hair color
creams.
t
t
t
Wet combing and color vibrancy test conducted on Caucasian hair tresses
Formulations tested contain Merquat polymers at 0.3% TS
The L value denotes lightness of shade, lower the L value darker the shade
CHROMAPOL 5 POLYMER
Create crystal clear gels and glossy creams that are gentle on the scalp. Deliver
vivid color while deeply conditioning your hair without the need to rely on
heavy dye loads. For uniform coverage and a new user experience choose NEW
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Gransil VX-418 offers

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C& T May 2 0 1 4
Ed it o r ' s n o t e | C&T
Cover art by James Fergus
2
4
6
96
Editor's Note
Regular Contributors
Scienti c Advisors
Advertiser Index
Market Intelligence
8
Turbulence in Emerging Markets

But Cosmetics Will Prevail by R. Walker
10
12
Finished Product Launches
14
Technology Launches
Read the Label: Ban Total Refresh

Cooling Body Cloths by S. Raffy
Regulatory
18 Lead, Aluminum and Parabens: Myths in
Cosmetics by C. Flower, PhD, and
E. Meredith, PhD
Research
22
Fatty Acid Bile Acid Conjugates Hypothesis

for Skin Anti-aging and Anti-acne Effects
by N. Dayan, PhD, and M. Halperin, MD
32
Understanding Fragrance Allergy,

Is Fragrance-Free Always Necessary?
by H.I. Maibach, MD, et al.
38
Molecular Biology in Future Skin and

Hair Care by H. Epstein, PhD
46
Review and Modern Advances of Retinoids

for Cosmetics by S. Isaacman, PhD, et al.
52
Patent Picks: Soft Hair Hold, Barrier Function

and More
Testing
56
Proposed Method to Evaluate the

Microbiological Stability of Cosmetics
During Use by N. Bresciani et al.
66
Correlating Aging with Skin's Mechanical and

Optical Properties by O. Freis, PhD, et al.
Formulating
76
Approaches and Issues in Powder

Formulations by P. Tsolis and G. Sahagun
84
Novel Azobenzene Compound to Extend and

Reactivate UV Protection by J.-Y. Wang, PhD,
and S. Geng
2 | www.CosmeticsandToiletries.com
Rachel L. Grabenhofer
NEXT-GEN COSMETICS
Years ago, Ken Klein reviewed an article submitted to Cosmetics
& Toiletries (C&T) on a cosmetic active. His feedback went
something like cosmetics should be cosmetic, not active, otherwise
they are drugs. He added that making such claims would draw the
scrutinizing eye of regulators. Many agree with this view. In fact,
the March 2013 cover of C&T featured an article on gene-silencing
for potential skin bene ts, and this stirred negative reactions from
readers. I wonder how the present cover fares?
To Klein's latter point, he was right. Several cosmetic
manufacturers have been issued warnings by the U.S. Food & Drug
Administration for making drug-like claims. To his rst point,
however, some argue cosmetics have always had e ects on the body.
In the April 2013 GCI, Steve Herman observed, Cosmetic science
used to resemble cooking. Now, it is looking like peer-reviewed
genetic research squeezed into a jar. He added, Some would say
that in reality, beauty products have always [altered genes] to a
certain extent. C&T takes no stand on what cosmetics should or
shouldn't be. We're here to provide you insight to envision what
they could be and this edition looks farther out than ever before
to research and technologies with real potential.
For one, topically impacting the human epigenome has been
all the buzz, and in this issue, Epstein introduces this concept and
its pathways into skin. In relation, Dayan and Halperin present a
hypothesis for a speci c fatty acid bile acid conjugate to a ect skin
via cellular membrane transporters.
Also looking to the future, Wang details how a avobenzene
compound attached to a cholesterol group has the potential for
sunscreens that can re-activate [Wang will present Reactivating
Sunscreens via Azobenzene Compounds at the Cosmetics &
Toiletries Summit (Summit.CosmeticsandToiletries.com) in June].
Freis et al. demonstrate the evolution of skin parameters with
aging, and identify a correlation between measured mechanical and
optical properties. And Bresciani et al. propose a method to assess
the antimicrobial e cacy of preservatives in cosmetics beyond
their containers i.e., during use.
Squeezing all this research into a jar may seem the work of
science ction, but it's of science fact. Even cosmetic cosmetics are
packing advanced particle science and optics into their compacts.
As knowledge and science continue to converge, who knows how
far next-generation products will reach.
Vol. 129, No. 4 | May 2014
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Regul ar Co n t r ibut o r s | C&T

Steven Isaacman, PhD, holds advanced
degrees in organic and physical organic
chemistry. He founded Nanometics LLC
in 2006, where he leads research on novel
molecules, polymers and materials for
personal care and pharmaceuticals.
p. 46
Howard I. Maibach, MD, is professor of
derma tology at the University of California
School of Medicine, San Francisco. His
laboratory has been interested in and
published extensively on dermatopharmacology and dermatotoxicology.
p. 32
Chris Flower, PhD, director-general of
the Cosmetic, Toiletry and Perfumery
Association (CTPA), joined CTPA a er
25 years in cosmetic safety and R&D.
He holds a doctorate from the CNAA in
toxicology and physiology/pathology.
p. 18
Susan Ra y is president of Susan Ra y
Consulting. She has more than 25 years
of experience in personal care, including
a number of roles in R&D, business
development and technical sales. She is an
active member of the SCC and ACS.
p. 12
Michael Isaacman, PhD, is a chemist

at Nanometics LLC and a postdoctoral
researcher at New York University. His
research is focused on the synthesis and
self-assembling dynamics of siliconebased amphiphilic block copolymers.
p. 46
Peter Tsolis has held various positions for
the past 14 years within e Est e Lauder
Companies R&D. He is an active member
of the Society of Cosmetic Chemists and
has presented on skin care formulation,
delivery systems and new technology.
p. 76
Emma Meredith, PhD, head of scienti c
and technical services at the CTPA, holds
a doctorate in pharmaceutical chemistry
from the University of Strathclyde. She is
particularly interested in sun protection,
hair colorants and cosmetovigilance.
p. 18
Nava Dayan, PhD, founded her research
consultancy a er 24 years in skin care.
She has written more than 150 articles
and four books, has been recognized for
excellence by the SCC and CRS, and was
awarded for innovation by In-Cosmetics.
p. 22
EDITORIAL
Editor in Chief
Jeff Falk
1-630-344-6071/ jfalk@allured.com
Editor
Rachel L. Grabenhofer
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Associate Editor
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1-630-344-6077/ kanderson@allured.com
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Fragrance
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CORPORATE
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Ot her products brought t o you by Allured:

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Vol. 129, No. 4 | May 2014
Scien t i c Ad vis o r y Bo ar d | C&T

Extreme multi-functionality is on the horizon; e.g., materials with anti-aging,
sunscreen, moisturizing and anti-free radical components in their structure, which
are slowly released to the skin, or that function continuously without being absorbed
in depth.
Luigi Rigano, PhD
Industrial Consulting Research
Eric Abrutyn
TPC2 Advisors
Ltd.
Prithwiraj Maitra,
PhD
Johnson & Johnson
David C. Steinberg
Steinberg & Associates
Angela R. Eppler,
PhD
P zer Consumer
Healthcare
Marc Pissavini,
PhD
Coty-Lancaster
Peter Tsolis
The Est e Lauder
Companies
Trefor Evans,
PhD
TA Evans LLC
Sylvianne
Schnebert, MD
LVMH Recherche
Russel Walters,
PhD
Johnson & Johnson
S. Peter Foltis,
PhD
L'Or al
G nther
Schneider,
PhD
Beiersdorf AG
Xiao Wu, PhD

Eli Lilly and Co.
Mindy Goldstein,
PhD
Atlantic Coast
Media Group
Ron Sharpe
Amway Corp.
Shuliang Zhang,
PhD
Unilever
Shuzo Ishidate,
PhD
Shiseido Research
Center
Leslie C. Smith,
PhD
Coty-Lancaster
Zoe Diana Draelos, MD

Dermatology
Consulting Services
Sunscreen is the most potent anti-aging product on the market today.
Vol. 129, No. 4 | May 2014
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LexFeel Natural
An extremely light and dry emollient
suitable as a natural alternative to
cyclomethicone and mineral oils.
Lexgard Natural
A mild, natural, multi-functional emollient/
humectant that assists in formulating
self-preserving cosmetics.
LexFeel N Series
Unique, natural uids that provide sensory
bene ts remarkably similar to silicones.
ideas start here
www.inolex.com
Mar ket In t el l igen ce | C&T
Turbulence in Em erging M arkets
But Cosmetics Will Prevail

Rob Walker, Euromonitor International
he emerging economies are going through

a painful structural readjustment, and, as
a result, the emerging middle class (a key
engine of growth for the beauty and personal
care industry over the last decade) is losing
some of its swagger.
In China, the combined pressures of a cooling economy and more aggressive competition
from homespun and South Korean brands are
squeezing all the big multinationals. Tellingly,
newly released data from Euromonitor International shows growth in China's beauty and
personal care market dropped below 9% last
year (at xed U.S. dollar values), its weakest
performance in two decades.
Market conditions are tougher still in
Brazil and India. In both cases, middle-class
consumers have been trading down across a
ra of beauty and personal care categories. e
negative tilt in market conditions in India has
taken the industry by surprise.
Po w e r Sh ift D e la ye d
Save to
My Library, a
new Web t ool.
According to the latest forecasts from

Euromonitor International, spending on beauty
and personal care in the emerging markets
will be higher than in the developed markets
by 2018. A year ago, this power shi had been
expected to take place in 2016, but the choppier
economic waters have held things back. e
key point, however, is that emerging markets,
overall, still present a myriad of opportunities
for growth into the medium and long term,
despite the trickier operating conditions.
Quick Look
n Growth in China's beauty and personal care market

dropped below 9% last year (at xed U.S. dollar values), its
weakest performance in two decades.
n In Brazil and India, middle-class consumers have been
trading down across beauty and personal care categories.
n Spending on beauty and personal care in emerging
markets was expected to overtake spending in developed
markets by 2016. Although this shift is expected to be
delayed until 2018, emerging markets, overall, still present
a myriad of opportunities for growth into the medium and
long term, despite the trickier operating conditions.
ere were also some strong individual emerging market

performances last year. For example, sales of beauty and
personal care in Indonesia climbed 16%, fueled by a booming
middle class in secondary cities such as Balikpapan, Borneo.
Across the emerging markets, the big challenge is in adapting
a planning strategy to the changing climate, and in tailoring
products and marketing to the speci c pro le of consumers.
Editor's note: To read the full market report, see the May 2014
issue of GCI magazine or visit www.GCImagazine.com.
Reproduction in English or any other language of all or part

of this article is strictly prohibited. 2014 Allured Business Media.
Vol. 129, No. 4 | May 2014
VISIT US
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Mar ket In t el l igen c e | C&T
Finished Product Launches

AMAZON BOTANICAL HAIRPOWDER
Pravana | www.pravana.com
Pravana's new Nevo brand Super Shape, Lived-In, Twist and Detail hair care products include citric acid
for shine, sorbital for nourishment and body, and PVP for control and texture for hair. Speci cally, the
Lived-In Powder Potion styling product changes from a powder to a pomade for application, helping to
provide additional texture and styling capabilities.
Ingredients: Lived-in Powder Potion: Water (aqua), Aluminum Hydroxide, Glycerin, Sodium
Carboxymethyl Starch, PVP, Methyl Methacrylate Crosspolymer, Sorbitol, Phenoxyethanol,
Fragrance (parfum), Citric Acid.
BERRYEXFOLIATOR
Michael Todd True Organics | www.michaeltoddtrueorganics.com
Michael Todd True Organics debuted Wild Berry Exfoliating Peel. Fruit enzymes from wild blueberry,
strawberry, grape and raspberry, as well as alpha and beta hydroxy acids, work to exfoliate skin in order to
smooth ne lines and surface wrinkles, and improve skin texture and tone. Aloe works to oxygenate and
detoxify the skin for an added antioxidant and antibacterial bene t.
Ingredients: Organic Aloe Leaf Juice, Isopentyldiol, Glycerin, Polysorbate 20, Lactic Acid, Rosehip
Seed Powder, L-Arginine, Decyl Glucoside, Salicylic Acid, Galactoarabinan, Mandelic Acid, Malic Acid,
Gluconolactone, Willow Bark Extract, Hawaiian Noni Fruit Extract, Bergamot Fruit Oil, Wild Blueberry
Fruit Extract, Grapefruit Extract, Grape Seed Extract, Raspberry Fruit Extract, Raspberry Seed Extract,
Cranberry Fruit Extract, Prune Fruit Extract, Cherry Fruit Extract, Wild Bilberry Fruit Leaf Extract,
Strawberry Fruit Extract, Rosa Canina Seed Powder, Blueberry Fiber, Ylang Ylang Flower Oil, Orange
Oil, Pentylene Glycol, Butylene Glycol, Ethoxydiglycol, Sclerotium Gum, Sodium Benzoate, Methyl
Hydroxyethylcellulose, Dehydroacetic Acid, Ethylhexglycine.
BLACKBERRYBODYSERUM
J.R. Watkins | www.jrwatkins.com
J.R. Watkins released an Anti-Aging Body Care System that includes a body wash, body cream, hand
cream and body serum. e Anti-Aging Body Serum utilizes the antioxidant power of cold-pressed
grape seed and blackberry seed oils, combined with plant-based moisturizers to leave skin more
hydrated. e serum also protects skin against oxidative damage and helps maintain skin's function.
Ingredients: Anti-Aging Body Serum: Vitis Vinifera (Grape) Seed Oil, Macadamia Ternifolia Seed
Oil, Prunus Armeniaca (Apricot) Kernel Oil, Persea Gratissima (Avocado) Oil, Rubus Fruticosus
(Blackberry) Seed Oil, Fragrance (parfum), Tocopherol, Ascorbyl Palmitate, Retinyl Palmitate,
Carthamus Tinctorius (Sa ower) Seed Oil, Aloe Barbadensis Leaf Extract.
Ingredients/claims are published as provided to C&T magazine by the manufacturers.
Vol. 129, No. 4 | May 2014
Botanical Oils.
Ultimate Beauty Rituals.
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Expert in natural ingredients, NATUREX is launching NAT oleis , a range of botanical oils of exceptional
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Let NAT oleis take you on a journey to discover all our oils.
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Read the Label

Ban Total Refresh
Cooling Body
Cloths
ANDIROBA NUT OIL

Yves Rocher | www.yvesrocherusa.com
Yves Rocher has released its Beautifying Dry

Oil with andiroba nut oil, which o ers a high
concentration of omega 9, to help nourish skin.
It is also dry to the touch for a pleasant sensory
application experience.
Ingredients: Isopropyl Palmitate,

Octyldodecanol, Coco-Caprylate/Caprate,
Ethylhexyl Palmitate, Ethylhexyl Stearate,
Hexyldecanol, Hexyldecyl Laurate, Carapa
Guaianensis Seed Oil, Sesamum Indicum
(Sesame) Seed Oil, Fragrance (parfum),
Tocopheryl Acetate, BHT.
ACRYLATEHAIRLIFTER
Not Your Mother's | www.nymbrands.com
Not Your Mother's has extended its hair care

line with Plump For Joy ickening Hair Li er,
a volumizing spray product including acrylates
to thicken hair and add body. Also, glycerin
helps to strengthen and add shine to hair.
Ingredients: Water, AMP-Acrylates/Allyl

Methacrylate Copolymer, Fragrance,
Glycerin, Polysorbate 20, Acrylates/
C10-30 Alkyl Acrylate Crosspolymer,
Aminomethyl Propanol, DMDM Hydantoin,
Methylparaben.
YOGURT BODYWASH
Dial | www.dialsoap.com
Dial introduced Frozen Yogurt Cooling Body

Wash, which uses yogurt proteins as a natural
skin conditioner to help skin retain moisture
while providing nourishment. e product
also delivers a clean rinsing experience and
uses menthol to provide a cooling sensation to
the skin.
Susan Ra y, Susan Ra y Consulting

e viewpoints expressed in this column are those of the author and
do not necessarily re ect those of Cosmetics & Toiletries.
Ingredients: Water, Sodium Laureth

Sulfate, Cocamidopropyl Betaine, PEG-8,
Fragrance, Glycerin, Polyquaternium-7, Menthol, Ananas
Sativus (Pineapple) Fruit Juice, Carica Papaya (Papaya)
Fruit Juice, Psidium Guajava Fruit Juice, Citrullus Lanatus
(Watermelon) Fruit Juice, Hydrolyzed Yogurt Protein,
Propylene Glycol, Styrene/Acrylates Copolymer, PEG-7
Glyceryl Cocoate, PEG-200 Hydrogenated Glyceryl
Palmate, Cocamide MEA, Tetrasodium EDTA, Sodium
Benzoate, DMDM Hydantoin, Citric Acid, Sodium
Chloride.
Vol. 129, No. 4 | May 2014
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2014 The Lubrizol Corporation.
Technology Launches
BIODEGRADABLEEXFOLIANT
HAIRSMOOTHING
Induchem has launched

a natural exfoliant in two
grades that are non-irritating
to sensitive skin and
biodegradable. Biogomm'age
UE (INCI: Cellulose (and)
Hydroxypropylcellulose
(and) Tocopheryl Acetate
(and) CI 77007) and Biogomm'age WD (INCI: Cellulose (and)
Hydroxypropylcellulose (and) Panthenyl Triacetate) are produced
with the company's Safe-scrub technology, enabling particles to selfdisintegrate upon use, thereby preventing skin irritation and avoiding
the skin redness typically seen a er exfoliation. is technology was
found to have the same exfoliation e ciency as conventional apricot
kernel fragments of the same size distribution.
Biogomm'age UE, with pro-vitamin B5, helps to regenerate skin, and
Biogomm'age WD, with vitamin E, maintains skin barrier function.
Each grade contains pigments in addition to the active that are available
in three sizes: 200 m, 400 m and 900 m.
e exfoliants are stable in any formula type, but are recommended
for: leave-on whitening creams, to remove pigmented cells; dandru
shampoos, without leaving residue on an oily scalp; facial cleansers;
exfoliating lotions for sensitive skin; leave-on anti-aging serums, to
stimulate epidermal renewal; scrubbing hand sanitizers; and more. e
exfoliants are also compliant with Chinese regulations.
www.induchem.com
Croda has launched an ingredient designed

to smooth hair cuticles that have been
damaged or li ed by bleaching or styling
processes. Crodabond CSA (INCI:
Hydrogenated Castor Oil/Sebacic Acid
Copolymer) is a conditioning treatment
that helps repair the damage caused by
at-home or salon chemical treatments.
e cuticle-sealing performance of the
copolymer is long-lasting, and its e ects
can be imparted over multiple shampoo
washes. Treatment with the products results in hair that is smooth,
conditioned and healthy-looking. In addition, the treatment
prevented future damage. e ingredient is recommended at
0.5-5.0% in hair care, daily conditioners, intense conditioning
treatments and shampoos.
www.croda.com
LIPOPHILICSOLUBILIZER
Mibelle Biochemistry has sourced single-cell algae that grows on

glaciers at 0 C to create its latest anti-aging active. Snow Algae
Powder (Pending INCI: Chlamydocapsa sp.-101 Extract (and)
Maltodextrin (and) Lecithin (and) Water (aqua)) mimics the e ects
of calorie restriction to improve the longevity of skin cells. e algae
adapt to their extreme habitat by changing pigment concentrations.
In addition, the production of secondary metabolites such as
biopolymers (gallerten), antifreeze glycoproteins, stress modi ers and
osmotically active amino acids and sugars help these extremophile
algae to survive in their habitat. e company produces snow algae
powder sustainably in a tailor-made bioreactor.
At the cellular level, the active protects and activates two key factors
of the caloric restriction pathway the Klotho longevity gene and
the AMP-activated kinase (AMPK) energy sensor, which together
lead to improved cellular defenses, oxidative stress resistance, cell
detoxi cation and repair. Results in the skin are the initiation of
collagen production, and rejuvenation of the dermal-epidermal
junction. Consequently, the skin barrier is reinforced while the skin
appears fresher and detoxi ed, and age spots become less visible. e
active is recommended at 2-3% in rejuvenating and repair formulas,
age-defense products, youth-protecting and promoting skin care, and
formulas to increase skin's longevity.
www.mibellebiochemistry.com
Evonik has created a naturally derived solubilizer

for the formulation of lipophilic ingredients, such
as emollients and natural oils, into clear cosmetic
products. In addition to its solubilizing ability, TEGO
Solve 61 (INCI: Polyglyceryl-6 Caprylate (and)
Polyglyceryl-4 Caprate (and) Polyglyceryl-4 Cocoate
(and) Polyglyceryl-6 Ricinoleate) has good skin feel and
imparts moisturizing bene ts. e solubilizer is made
from 100% renewable raw materials and complies with
Ecocert standards.
Being easy to handle and cold processable, the ingredient allows for
the preparation of mixtures with oils and water. For example, fatty oils
such as avocado, olive, jojoba, sun ower and argan, as well as caprylic/
capric triglyceride, can be incorporated into clear formulations. e
product was found to outperform PEG-40 hydrogenated castor oil in
solubilizing natural oils and in vivo, in a short-term moisturization test
using a corneometer, the solubilizer was found to act as a humectant
and provide moisturization from a body lotion. e solubilizer is
recommended at 0.5 10.0% in shampoos, body/hand/facial wash and
gel, makeup remover, emulsions and wet wipes.
personal-care.evonik.com
ANTI-AGINGALGAE
Vol. 129, No. 4 | May 2014
Reen g in eer ed
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Pr emiu m S po n s o r s :
Regul at o r y | C&T
Lead, Alum inum and Parabens:
Myths in Cosmetics
Chris Flower, PhD, and Emma Meredith, PhD
Cosmetic, Toiletry and Perfumery Association, London, UK
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here seem to be plenty of myths surrounding cosmetic products

and their ingredients. e rst of these is that a manufacturer would
deliberately use an ingredient that might lead to harm. Apart from
being illegal, it is not good for business to injure consumers, and successful
companies understand the needs of their consumers. is column will look
at some of the stories that are propagated online, and try to separate fact
from ction, as there is an element of both in all good myths.
Le a d a nd Lip s t ic k
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e rst is that perennial favorite lead in lipstick. It has reappeared

several times in di erent guises, and has even been prompted by well-meaning analysis of products by reputable agencies. Delving rst into the facts,
lead is harmful, but anything can be harmful if consumed excessively. ere
is no known biological use for lead in humans, so it is best avoided. But can
lead be avoided? Not completely, is the honest answer. Recognizing that lead
is harmful and not useful, lead has been removed from petrol (gasoline),
(most) paints, the solder used to seal cans for food, and household water
pipes. However, lead is present naturally in the environment, where it is
absorbed by plants from the soil, and is unavoidable in root vegetables.
Furthermore, because lead is ubiquitous, it is also found at trace levels in
some substances used to make products such as lipstick. It is not added
deliberately, but it cannot be completely avoided. e question is whether
the amount of lead in lipstick might be harmful to consumers, and the
answer is a clear no. In fact, Health Canada has said that people get more
lead from safe drinking water than the tiny traces present in lipstick.
is lead in lipstick myth is naturally related to two more myths that
women consume a given quantity (the exact amount varies with the
di erent stories) of lipstick in a lifetime, and that an individual can tell if
a lipstick contains lead by rubbing it on a gold ring. ink of how many
lipsticks a woman purchases per year, how much is le unused or discarded,
and how much is le on tissues, drink ware and even the faces of friends
and family. One soon realizes that the estimated tubes of lipstick that a
woman consumes is, at best, an exaggerated guess.
As for the gold ring test, it is sheer nonsense. It takes the nest of
analytical chemists using the latest sophisticated equipment to measure tiny
traces of lead.
Vol. 129, No. 4 | May 2014
Visit us at booth #331, NYSCC Suppliers' Day

Edison, New Jersey, May 13 - 14 2014
CON N ECTED
BY CH EM I STRY
ALL THE INGREDIENTS
FOR THE ULTIMATE
FORM ULATION
With a shared commitment to creative
innovation, Innospec has integrated
Chemsil into its personal care portfolio to
offer an unrivalled range of customizable
solutions.
Chemsil's high-quality speciality silicones
complement Innospec's extensive range of
personal care ingredients to deliver creative
formulation solutions. With a uniquely customercentric approach, we work in close collaboration
with formulators to support the development of
exciting products with real consumer appeal.
We can help you gain a market edge across
personal care categories, including:
Hair care
Skin care
Sun care
Intimate care
For inspiration, ideas or further information,
please contact us.
w w w.innospecinc.com
AMERICAS : america-pc@innospecinc.com ASIA-PACIFIC : aspac-pc@innospecinc.com
EUROPE, MIDDLE EAST & AFRICA : emea-pc@innospecinc.com
Regul at o r y | C&T
e cosmetic industry, however, should not be complacent. ere are,
unfortunately, some instances of illegal cosmetic products being discovered on
the market that are extremely high in lead, since in these cases, lead is one of the
main deliberate ingredients. e fact that such concoctions have been used in
some communities as traditional decorative products for generations does not
lessen the risk to those using them. ese are illegal cosmetics and should be
dealt with accordingly.
Pa r a b e n s
Another myth surrounds the safety of parabens. ere have been reports that
these preservatives are linked to breast cancer through an ability to mimic the
female hormone estrogen. Here, the facts are clear. Parabens are used as preservatives to ensure that cosmetics remain wholesome and safe throughout their
use and do not have to be discarded quickly. Parabens are found in nature; many
fruits contain parabens made by the plant itself to prevent the fruit from molding. Some parabens are able to mimic a portion of the properties of estrogen, but
not all of them. at mimicry is only seen under experimental conditions with
very high exposure or doses, and such conditions do not relate to everyday life.
Indeed, it is perfectly impossible for a human being to be exposed to su cient
parabens from cosmetic products to ever produce any disruption of the hormone
system and even parabens found to mimic some properties of estrogen are
poor copies. Of course, many remember the study that claimed to have found
parabens in breast cancer tissues, but this study was poorly conducted and has
been strongly criticized by scientists. Parabens from an unidenti ed source had
contaminated many of the samples, including the blank controls, which should
not have any parabens present.
A lu m in u m in An t ip e r s p ir a n t s
More recently, the cosmetic industry has seen concerns raised about the safety
of aluminum in antiperspirants. Aluminum is the most abundant metal on earth,
and is the third most abundant element. If it were particularly toxic, life itself
could not exist in its presence. In fact, aluminum has no known biological function in humans, and what humans absorb is readily removed via the kidneys.
It can cause harm when present in excess, which happens when kidneys
malfunction and when exposure is excessive with aluminum-based antacids or
during work in the aluminum industry. Any contribution from antiperspirant
use is small, particularly considering that aluminum compounds remain on the
surface of the skin to function by forming a gel to plug the sweat ducts. If the
aluminum was absorbed into the skin, the product would no longer work.
C o n c lus io n
ere are plenty more myths that have come and gone, but they all have
certain things in common: information is either exaggerated or not placed into
context, and incomplete information causes consumers to question where, in
fact, no problem exists. In all of these discussions, please remember that reputable cosmetics companies not only comply with strict legislation but want to
build a long-lasting relationship with their consumers, to have those consumers
stay loyal to the brand, and to make repeated purchases. is will not happen if
the company fails to provide satisfaction in terms of safety, e cacy and quality.
Vol. 129, No. 4 | May 2014
Introducing Perlaura
Add more life to your skin care regimen with Perlaura
the new anti-aging active for visibly brighter, smoother skin.
Just one of many new ingredients you will nd in booth # 213
at the NYSCC Suppliers Day on May 13 & 14.
carecreations.basf.com
Res ear ch | C&T
Fatty Acid Bile Acid Conjugates
Hypothesis for Skin Anti-aging

and Anti-acne Effects
Nava Dayan, PhD
Dr. Nava Dayan LLC, NJ, USA
Maya Halperin, MD
Galderm Therapeutics, Tel Aviv, Israel
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atty acid bile acid conjugates (FABACs) are a family of small synthetic
molecules that initially were developed as oral drugs to reduce fat
build-up and accumulation in the liver. e structure-activity rationale
is that the saturated fatty acid acts as a cholesterol solubilizing agent while
the bile acid acts as a vehicle to enable secretion into bile and penetrate into
the enterohepatic circulation. e amide bond further enhances stability
against intestinal degradation.1 In the skin, however, cholesterol metabolism
di ers dramatically. Skin renewal is maintained by controlling the balance
between proliferation, di erentiation and apoptosis of epidermal cells,2
and it has been shown that this program of epidermal di erentiation in
keratinocytes is altered when cholesterol-enriched domains in the plasma
membrane are disrupted.
Leveraging the innovation of FABACs in health care, the authors
developed a speci c FABACa based on a cholesterol-solubilizing moiety,
i.e., saturated fatty acid, and a bile acid (cholic acid) as the vehicle to enable
secretion into bile and entry into the enterohepatic circulation for potential
skin bene ts.3, 4 is compound was chosen for its relatively low molecular
weight and lipophilicity, allowing it to penetrate skin, a ect cholesterol
on the cell membrane level and facilitate other mechanisms. Previous
proteomic data has proven the activities of FABACs5-7 in enhancing
ATP-binding cassette (ABCA1) cholesterol transporter and competitively
inhibiting stearoyl-CoA desaturase (SCD1) enzyme. erefore, it was
hypothesized that the developed FABAC would a ect skin in similar ways.
In this paper, the mechanisms of ABCA1 cholesterol transporter and
SCD1 enzyme in the skin are detailed rst, highlighting the structure-activity relationships (SARs) involved. Following this, in vitro screenings of the
FABACa active are described; screenings determined the level that activity
was occurring. Interestingly, compiled results suggest activities comparable
to retinoic acid the only drug currently prescribed for skin aging and
known for anti-acne e ects (see Page 46 for more on this ingredient).
However, retinoic acid acts through nuclear receptors, whereas the new
FABAC is believed to act on cellular membrane transporters and competitively inhibit enzymes by depleting cholesterol from the membrane, thereby
changing membrane uidity and the exposure of membrane-anchored
a
Steamchol, Galderm
erapeutics

Vol. 129, No. 4 | May 2014
s
s
s
s
Res ear ch | C&T

receptors. is milder yet e ective mode of action is
an attractive option due to its larger margin of safety.
C h o le s t e r o l in Sk in
e skin is a site of active lipid synthesis. In the
stratum corneum, aliphatic lipids are synthesized
de novo in the epidermis via phospholipids, and
cholesterol is synthesized from acetate within hours
a er induction (see Figure 1); cholesterol esters are
produced three to seven days later. e skin's lipid
pro le a ects its ability to serve key functions, such
as acting as a barrier against insult and preventing
water loss from the body.
Skin b arrier re new al

invo lves the g e neratio n
and sec retio n o f lam ellar
b o d ies into the ECM .
Cholesterol is the second most abundant lipid
in the stratum corneum a er ceramides which
account for up to 50% w/w of total intercellular lipids.
Cholesterol is known to promote the intermixing of
di erent lipid species and to regulate their thermodynamic phase behavior. Cholesterol esters, i.e., the next
step in the synthesis chain, contain unsaturated and
saturated diacyl chains that contribute to the stratum
corneum's stability and uidity, and promote its liquid
condensed state. In these capacities, both cholesterol
and its esters serve a fundamental role in skin's lateral
lipid organization, and their ratio controls skin barrier properties.8
Since skin stem cells undergo terminal di erentiation, and skin acts in some aspects as a separate
entity, as several compartments in the skin are not in
equilibrium with the body's circulation, and, therefore, it ful lls its own needs from the blood. Further,
the metabolic pathways of skin lipids are di erent

from the pathways in internal organs and blood;
consequently, there is no correlation between the
cholesterol levels in blood and skin.
e skin barrier renewal process involves the
generation and secretion of lamellar bodies or
granules from keratinocytes into the extracellular
matrix (ECM), which requires utilizing a battery
of enzymes.9 ese lamellar bodies contain lipid
precursors such as glucosylceramides, cholesterol,
glycerophospholipids and sphingomyelin, as well
as catabolic enzymes such as proteases, lipases, acid
phosphatase and glucosidases. Such lipid precursors
are metabolized in the ECM and fused end-toend, forming progressively elongated membrane
sheets the intercellular lipid lamellar structure of
the stratum corneum.
Under basal conditions, lamellar body secretion is
relatively slow and corresponds to the kinetics demonstrated by Hedberg and Wertz.10 However, when
acute insult to the barrier is applied, this process is
accelerated to promote a sequence of recovery that
is achieved a er 72 hr in young skin (i.e., 20s/30s).
is sequence includes increases in cholesterol, free
fatty acids and ceramide synthesis, all of which are
restricted to the underlying epidermis at the injured
site and dependent upon a prior up-regulation of
mRNA encoding to synthesize anabolic enzymes.
Since the synthesis of each of these key lipids is
essential for maintaining normal barrier properties,
inhibiting their synthesis may lead to abnormalities
and an impaired barrier that is more permeable.
C h o le s t e r o l Re g u la t io n ,
A B C A 1 a n d SC D 1
Keratinocytes require abundant amounts of
cholesterol for maintaining a strong barrier and
controlling cutaneous permeability; hence, the
regulation of cholesterol homeostasis in the skin
is of great importance. ABCA1 plays a pivotal role
for cholesterol e ux. It regulates cholesterol levels
by promoting the transport of cholesterol and
Figure 1. Synthesis path o f c holesterol in skin
HMGCoA
Synthase
Acetate
HMGCoA
Reductase
HMGCoA
Squalene
Synthase
FPPS
Melvalonate
Farnesol
Squalene
Enzyme
Cholesterol
Substrate
Vol. 129, No. 4 | May 2014
phospholipids across cell membranes. Jiang et al.11

demonstrated the expression of ABCA1 in human
keratinocytes and murine epidermis, con rming its
localization both in the outer epidermis, i.e., stratum corneum and stratum granulosum, and lower
compartments including the stratum spinosum and
stratum basale. e activation of ABCA1 was shown
to lead to the activation of the retinoid X receptor in
keratinocytes and in macrophages.12
Jiang et al. also demonstrated that acute disruption of the barrier by tape-stripping or the application
of acetone increases the synthesis of cholesterol and
suppresses the expression of ABCA1 transporter;
that way, cholesterol remains local and is available for
rapid barrier repair. Conversely, facilitating ABCA1
activity results in transport of cholesterol into the cell,
reducing the presence of cholesterol in the keratinocyte membrane and stratum corneum. is suggests
the ABCA1 transporter may be linked to keratinocyte
di erentiation.
In relation, studies13 point toward a cascade
connection between stearoyl-CoA desaturase (SCD1)
enzyme inhibition and ABCA1 activation in skin.
Mice de cient in SCD1 demonstrated sebaceous
gland atrophy, depletion of sebaceous lipids, dry
skin and alopecia. Interestingly, researchers suggest
that the deletion of SCD1 and resulting reduction
in sebaceous lipids may be of value in the treatment
of Acne vulgaris, which is associated with increased
sebaceous gland activity.
C h o le s t e r o l a nd Lip id Ra ft s
A closer investigation of cholesterol points to its
indirect role in controlling cell cycles. It accomplishes
this via receptors and transporters at the cell membrane, a ecting the organization of protein-binding
functions, in turn changing the protein conformation
and interacting with nearby clusters. e plasma

membrane of cells is made up of a combination of
glycosphingolipids and protein receptors organized
in glycolipoprotein micro-domains referred to as
lipid ra s (see Figure 2). ese specialized entities
compartmentalize cellular processes by organizing
the assembly of signaling molecules, in uencing
membrane uidity and tra cking membrane proteins. Lipid ra s are more ordered and tightly packed
than the surrounding and relatively uid bilayer but
oat freely in the membrane bilayer. ey have been
described as small (10-200 nm), heterogeneous,
highly dynamic, sterol and lipid enriched domains.
Further, small ra s can sometimes be stabilized to
form larger platforms through protein-protein and
protein-lipid interactions.14
One key di erence between lipid ra s and the
plasma membrane from which they are derived is the
lipid composition. Research has shown 15 that lipid
ra s generally contain three to ve times the amount
of cholesterol found in the surrounding bilayer.
Cholesterol interacts preferentially, although not
exclusively, with sphingolipids due to their structure
and the saturation of the hydrocarbon chains. So
although not all of the phospholipids within the ra
are fully saturated, the hydrophobic chains of the
lipids contained in the ra s are more saturated and
tightly packed than the surrounding bilayer. Cholesterol can be viewed as the dynamic glue that holds
the ra together.
Due to the rigid nature of the sterol group,
cholesterol partitions into the lipid ra s where the
acyl chains of lipids tend to be in a less uid state.
One important property of membrane lipids is their
amphiphilic character having a polar, hydrophilic
head group and a non-polar, hydrophobic region.
Cholesterol has the ability to pack in between the
Figure 2. Illustration of lipid raft; reg ion 1) is standard lipid bilayer w hile region 2) is a lipid raft
* 1) Non-raft membrane, 2) Lipid raft, 3) Lipid raft associated transmembrane protein, 4) Non-raft membrane protein, 5) Glycosylation
modi cations (on glycoproteins and glycolipids), 6) GPI-anchored protein, 7) Cholesterol, 8) Glycolipid
Vol. 129, No. 4 | May 2014
Cosmetics & Toiletries
| 25
Res ear ch | C&T

lipids in ra s, serving as a molecular spacer and lling
any voids between associated sphingolipids. e depletion
of cholesterol from lipid ra s has been shown to change
their organization and a ect keratinocyte di erentiation. It
was demonstrated, for example, that when lipid ra s were
treated with low concentrations of methyl-beta cyclodextrin, which entrapped cholesterol and removed it from
the ra , a signi cant decrease in keratin 1 and 10 early
di erentiation markers was observed.16
M a t e r ia ls a n d M e t h o d s
Based on the biology and SARs, as stated previously, it
was hypothesized that the speci ed FABAC would enhance
ABCA1 cholesterol transporter and competitively inhibit
SCD1 enzyme. us, its e ects were assessed in vitro via a
gene expression assay, described here.
Full thickness model: Gene expression was measured in
a full-thickness skin culture modelb. e FABACa ingredient was applied to the surface of each test culture at a
concentration of 0.5% and collected 24 hr post-application.
Cultures treated with 10 mL 100% DMSO served as the
vehicle control group. Tissues were collected in an RNA
stabilization solution c, and gene expression was analyzed
using validated assaysd. A set of 94 genes known for functions in the skin were evaluated, including ABCA1 and
SCD1 genes. Statistics were carried out using so waree.
Re s u lt s a n d D is c u s s io n
Of the 94 selected genes used in the panel, only
two keratin 1 and 10 (KRT 1 and 10) demonstrated
statistically signi cant deviations in expression (see
Table 1). ABCA1 and SCD1 mRNas were not altered
by the FABAC at the gene expression level as expected.
However, considering the SARs and previous proteomic5-7
data, these ndings, in fact, support the theory of e ects
being con ned to the protein level. Further, the FABAC
was speculated to be depleting cholesterol levels in lipid
ra s. As noted, the depletion of cholesterol from lipid ra s
was shown to change their organization and signi cantly
decrease keratins 1 and 10; here, KRT 1 and 10 were
reduced. To elucidate the receptors being a ected further,
additional studies are planned.
b
Epiderm FT, Mattek

RNAlater, AMBION, Inc.
d
TaqMan, Roche Molecular Systems, Inc.
e
StatMiner so ware v4.2, Integromics Inc.
c
Table 1. Statistically Signi cant Fold-c hange

Data for 0.5 % FABAC a vs. DM SO Control
Gene ID
Gene name
Fold-change
KRT10
keratin 10
-2.6
KRT1
keratin 1
-2.49
Vol. 129, No. 4 | May 2014
K e r a tin s 1 a n d 10
To further understand the e ects of the specied FABAC, consider the role of keratinocytes.17
Keratins are heteropolymeric structural proteins
that form the intermediate lament. ese laments,
along with actin micro laments and microtubules,
compose the cytoskeleton of epithelial cells. e
intermediate laments are assembled from keratin
monomers, and the corni ed envelope is assembled
from a protein called involucrin, as well as others.
Involucrin is synthesized in the stratum spinosum,
where it is cross-linked by transglutaminase enzyme,
which further stabilizes it. us, involucrin provides
structural support to the cell and allows for resistance
to microorganisms. Involucrin also binds to loricrin,
another protein, and contributes to the formation of
the corni ed envelope.
Keratins 1 and 10 are heterodimers and major
constituents of the intermediate lament cytoskeleton in the superbasal epidermis. Both keratins
are expressed in the spinous and granular layers
of the epidermis. Type II cytokeratins consist of
basic or neutral proteins that are arranged in pairs
of heterotypic keratin chains co-expressed during
the di erentiation of simple and strati ed epithelial
tissues. e down-regulation of keratins 1 and 10 has
been associated with the up-regulation of involucrin, and both have been shown to be triggered by
exposure to retinoic acid.18
e up-regulation of involucrin production, as
a result of the down-regulation of keratins 1 and
10, can therefore be explained as a compensation
mechanism that allows the epidermis to maintain
its integrity in spite of the attenuated di erentiation.
Table 2 cross-sections the normal human skin, outlining its layers, corresponding cell types, expressed
keratins and other markers. It should be noted that
the expression of keratins 1 and 10 is linked to the
expression of involucrin and con ned to the epidermal spinous layer in which critical biochemical
paths that determine the integrity of the barrier and
its appearance are found.
Following the expression of keratins 1 and 10,

pro- laggrin, an important marker, is expressed and
leads to the generation of laggrin, a cationic protein
speci c to the stratum corneum. Taken together,
in theory, the FABAC's attenuation of keratinocyte
di erentiation and depletion of cholesterol from lipid
ra s could lead to a compensation mechanism that
results in accelerated proliferation and barrier rejuvenation, potentially renewing the skin and a ecting its
appearance. Additional studies are under way.
Re t in o ic A c id , a n d
K e r a t in s 1 a n d 10
Retinoic acid is the active form of vitamin A. e
four most common indications for retinoids are:
acne, wrinkles, photo-damaged skin and inheritable
keratiopathies however, the potential for teratogenic e ects from the use of retinoids in women
of children-bearing age is a key consideration.19 In
addition, it is well-established that a common adverse
e ect from retinoid treatments is skin irritation,
although the exact mechanism is not fully elucidated.
e general hypothesis is that retinoids normalize
keratinocyte di erentiation; other possible mechanisms include the down-regulation of desmosomal
proteins, anti-proliferative e ects, regulating lipid
synthesis, growth factors and cytokines.
Unlike the theoretical mechanism of the new
FABACa, retinoids exert their e ects entirely through
nuclear receptors. ere are at least six retinoic acid
receptors belonging to two families: retinoic acid
receptors (RARs) and retinoid X receptors (RXRs).
Nuclear receptors expressed in keratinocytes include
these two families of receptors, vitamin D3 receptor
and thyroid hormone receptor. All of these can a ect
keratinocyte di erentiation. For example, the RAR
gamma receptor for retinoic acid plays an important
role in the morphogenesis and di erentiation of
squamous epithelia.
Retinoic acid has been shown to inhibit the
activity of keratinocyte transglutaminase and the
formation of the corni ed envelope. Similar to the
Table 2. Norm al Hum an Skin Layers, Corresponding Cell Types, Expressed Keratins
and Other M arkers
Skin sub tissue
Cell type
Expressed keratins
Other markers
Stratum corneum
Corneocyte
Stratum granulosum
Granular keratinocyte
Keratin 2
Stratum spinosum
Spinous keratinocyte
Keratins 1 and 10
Involucrin
Stratum basale
Basal keratinocyte
Keratins 5 and 14
Integrins
Stratum dermis
Fibroblast
None
Collagen and elastin
Vol. 129, No. 4 | May 2014
Corni ed envelope
Loricrin, laggrin
| 27
Res ear ch | C&T

FABACa, both keratins 1 and 10 were shown to be
down-regulated in skin treated with retinoic acid
in vitro and in vivo.20 Interestingly, however, not all
di erentiation markers are regulated by retinoic acid;
involucrin, for example, is una ected when cultured
keratinocytes are treated by retinoic acid (see Page 46
for further discussion).
SC D 1 a n d A n t i- a c n e Po t e n t ia l
As noted previously, studies11 point to a connection between SCD1 enzyme inhibition and ABCA1
activation in skin; the deletion of SCD1 reduces
secretion of sebaceous lipids, which may be of value
in the treatment of acne. In relation, the indirect
inhibition of SCD1 activity by FABACs does not
appear to a ect barrier integrity, unlike retinoic
acid. is is believed to be a result of the compensation mechanism. While inhibition of SCD1 leads
to an increase in ABCA1 transporter activity and
the depletion of barrier cholesterol levels in keratinocytes, a compensatory enhanced production of
cholesterol and ceramides may lead to a replenished
barrier. is restored barrier is especially necessary in
the case of reduced sebum secretion, which acts as a
protecting layer.
ere are numerous factors for the initiation and

progression of acne, and its severity can be related
to the interplay between these factors. Table 3 and
Figure 3 summarize the physiological conditions
leading to acne and the anticipated activity of the
speci ed FABACa based on its structure, SARs with
similar FABACs, and data generated from the gene
expression study.
Pr a c t ic a l A p p lic a t io ns
e presented active has potential for anti-aging
and anti-acne applications, and the next steps are to
study its behavior in topical formulations and acquire
safety assessment data relevant to the skin. When
considering a compound for potential biological
activity, rst-tier studies should seek to understand
its mode of action (i.e., pharmacodynamics) and
site of action i.e., the skin sub-tissue and cellular,
receptor and enzyme levels. Based on this screening,
a hypothesis is drawn, which is the stage at which
the present authors have arrived and at which this
paper was written. Perhaps the most interesting
aspect is that FABACs impart biological activity
manifestations similar to those of a known drug but
by a ecting di erent cellular entities.
Table 3. Conditions Lead ing to Acne and Anticipated Activity of the Speci ed FABAC
Abnormality in acne
Clinical/physiological effect
Potential FABAC activity to attenuate

the condition
Increased sebum production

as a result of elevated blood
hormone levels or other
abnormal physiology
Sebum serves as a nutrient,

fostering bacterial
proliferation in the gland
Potential SCD1 enzyme* inhibition,

associated with:
a) Depletion in sebum lipids
b) Attenuation in sebaceous gland activity
Abnormal microbiota
population, especially
Propionobacterium acnes
(P. acne)
P. acne consumes sebum

and over-proliferates,
resulting in the release of
in ammatory fatty acids and
toxins
Due to its chemical properties, lipophilicity

and relatively low MW, the FABAC may
intercalate into the bacterial wall, create
imbalance and partial perturbation of the
barrier, and therefore decrease the bacteria
population
Corni cation of the

pilosebaceous duct that
narrows the duct opening
and restricts sebum
drainage
Obstruction of sebum ow
from the gland onto the
surface, therefore appropriate
drainage of the gland is not
achieved
Attenuation of keratinocyte differentiation

to corneocytes; relief of pore-clogging of
the pilosebaceous opening, allowing more
effective sebum drainage**
In ammation
Humoral immune response

to free fatty acids and
toxins; recruitment of
in ammatory immune cells
and biomarkers that leads
to skin damage and can
cause scarring
Depletion in sebum lipid and potential

antimicrobial activity can lead to reduction
in in ammation
* Stearoyl CoA-Desaturase-1 (SCD-1) is an enzyme that catalyzes the synthesis of monounsaturated fatty acids from saturated fatty acids.
** Such an effect is claimed for both retinoic acid and salicylic acid
Vol. 129, No. 4 | May 2014
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Figure 3. Diagram desc ribing the hypothesis for FABAC anti-acne activity
Affecting arachidonic
acid cascade
Reduction in
in ammation
Attenuation in conversion
of saturated to mono
unsaturated fatty acids
FABAC inhibits
SCD1 enzyme and
enhanced ABCA1
transporter activity
Induction of changes
in sebum composition;
less wax diesters and
triglycerides
Reduction in sebaceous
gland activity
Activation of ABCA1
transporter
Potential anti-microbial
effect
Reduction in p. acne
population
Potential compensation
mechanism of RXR
activation
A retinoic acid like

anti-acne effect
Down regulation of
keratin 1 and 10
Decrease in corni cation

of the pilosebaceous
gland duct
Su m m a r y
5. US Pat 6384024B1, Bile salt conjugates, assigned to T Gilat

(May 7, 2002)
Here, the authors present the potential activity of

a selected FABAC on skin. Initial in vitro screenings
are presented, leading to a hypothetical suggested
activity of the active. Based on screenings, SARs and
the ingredient's proposed mechanism of action, a
hypothesis for its activity as an anti-aging and antiacne active is drawn.
Interestingly, its suggested mechanism can be
compared to that of retinoic acid, but while retinoic
acid acts via the activation of nuclear receptors, the
FABAC is thought to act on a cellular membrane
transporter level and through competitive enzyme
inhibition. Lastly, this apparently milder yet e ective mode of action could translate to larger safety
margins; when tested for cytotoxicity on a full thickness model, the FABAC demonstrated no signi cant
changes in cellular viability up to levels of 2%.
6. US Pat 6395722B2, Fatty acid derivatives of bile acids and bile

acid derivatives, assigned to T Gilat (May 28, 2002)
Acknowledgments: Galderm erapeutics, an Israeli start-up company,

and Nava Dayan, PhD (nava.dayan@verizon.net), collaborated on the
present work; the authors are thankful to Prof. Philip W. Wertz, from the
University of Iowa, for his critical review of this paper.
References
1. I Goldiner et al, ABCA1 dependent but apo A-1 independent
cholesterol ef ux mediated by fatty acid-bile acid conjugates
(FABA's), J Biochem 396 526-536 (2006)
7. US Pat 6589946B2, Bile salt conjugates, assigned to T Gilat (Jul

8, 2003)
8. I Kravchenko, Y Boyko, N Novikova, A Egorova and S Andronati,
In uence of cholesterol and its esters on skin penetration in vivo
and in vitro in rats and mice, Ukrainica Bioorganica Acta 1 (2011)
17-21
9. MR Prausnitz et al, Skin barrier and transdermal drug delivery,
available at drugdelivery.chbe.gatech.edu/Papers/2012/Prausnitz%20Derm%20Book%20Chapter%202012.pdf (Accessed
Mar 21, 2014)
10. CL Hedberg, PW Wertz and DT Downing, The nonpolar lipids of
pig epidermis, J Inves Derm 90 225 229 (1988)
11. YJ Jiang, B Lu, PM Elias and KR Feingold, Regulation of ABCA1
expression in human keratinocytes and murine epidermis, J
Lipid Res 47 2248-2258 (2006)
12. P Costet et al, Retinoic acid receptor-mediated iInduction of
ABCA1 in macrophages, Mol Cell Biol 23 (21) 7756-7766 (2003)
13. Ibid Ref 10
14. S Lambert, R Gniadecki, and Y Poumay, Cholesterol and lipid
rafts as regulators of signaling through the EGF receptor in
keratinocytes, Open Dermatology J 3 151-158 (2009)
15. LJ Pike, Lipid rafts, bringing order to chaos, J Lipid Res 44,
655-667 (Apr 2003)
16. F Sporl et al, Real-time monitoring of membrane cholesterol
reveals new insights into epidermal differentiation, J Invest Derm
130(5) 1268-1278 (2010)
17. RL Eckert and ER Rorke, Molecular biology of keratinocytes
differentiation, Environ Health Perspect 80 109-116 (1989)
2. J Nie, X Fu and W Han, Microenvironment-dependent homeostasis and differentiation of epidermal basal undifferentiated
keratinocytes and their clinical applications in skin repair, J Eur
Acad Dermatol Venereol 27(5) (2013) 531-535 (2013)
18. Y Poumay, F Herphelin, P Smits, IY De Potter and MR Pittelkow,

High-cell density phorbol ester and retinoic acid upregulate
involucrin and downregulate suprabasal keratin 10 in autocrine
cultures of human epidermal keratinocytes, Mol Cell Biol Res
Commun 2(2) 138-144 (1999)
3. T Gilat et al, Fatty acid bile acid conjugates (FABACs) New

molecules for the prevention of cholesterol crystallization in bile,
Gut 48(1) 75-9 (Jan 2001)
19. C Fisher, M Blumberg and M Tomic-Conic, Retinoid receptors

and keratinocytes, Crit Rev Oral Biol Med 6(4) 284-296 (1995)
4. T Gilat et al, Arachidyl amido cholanoic acid (Aramchol) is a

cholesterol solubilizer and prevents the formation of cholesterol
gallstones in inbred mice, Lipids 36(10) 1135-40 (Oct 2001)
20. H Torma, Regulation of keratin expression by retinoids, DermatoEndocrinology 3:3 136-140 (2011)
Vol. 129, No. 4 | May 2014
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Understanding Fragrance Allergy

Is Fragrance-free Alw ays Necessary?
Howard I. Maibach, MD
University of California San Francisco, San Francisco, USA
Garrett Coman and Nicholas Blickenstaff

University of California San Francisco, San Francisco, USA;
and University of Utah, Salt Lake City, USA
Ashley Edwards
Touro University, Vallejo, CA, USA
KEY WORDS
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ragrances have become ubiquitous in skin and hair care products

to appeal to the consumer's senses. However, fragrances have been
associated with allergic contact dermatitis in applications including:
unspeci ed leave-on products, sun tan lotion,1 deodorants,2 scented lotion,
unspeci ed rinse-o products, ne fragrances, shampoo, liquid soap, a ershave, lipstick, sunscreen, hair styling products, shaving foam, mascara, hair
dye, eye shadow and makeup cream. An example of typical allergic contact
dermatitis of the axilla due to a deodorant fragrance is shown in Figure 1.
While some dermatologists recommend avoiding all fragrances yielding
positive patch test results, as shown in Figure 2, it has become increasingly
di cult to avoid all fragrances and in the end, may be unnecessary for the
patch test positive patient.
To identify dermatitis caused by fragrance, a patch test3 for common
aromatic allergens was designed in which two fragrance mixes using
putative common allergens served as screens. Fragrance Mix #1 (FM1) was
developed from the fragrances used in an antifungal cream that had caused
an allergic contact dermatitis epidemic.4 It comprised: Evernia prunastri
(oak moss), isoeugenol, cinnamyl alcohol, eugenol, cinnamal, geraniol,
-amylcinnamal and hydroxycitronellal. Fragrance Mix #2 (FM2), a later
attempt5 to identify fragrance allergens, consisted of hydroxyisohexyl
3-cyclohexene carboxaldehyde (HICC), farnesol, citral, hexyl cinnamal,
citronellol and coumarin. ese materials are considered EU fragrance allergens; Table 1 on Page 36 lists them with their FM designations, along with
additional EU fragrance allergens. ese mixes have been used in fragrance
allergy testing reported in the literature, which are reviewed here.
Pr e lim in a r y Q u e s t io n n a ir e
e diagnosis of a fragrance contact allergy always starts with a detailed
medical history. Schollhammer et al. developed a questionnaire to determine if consumers had a certain, probable or possible allergy to fragrances
based on their recollection of adverse reactions to perfumes or perfumed
products.6 In this questionnaire, the certain allergy included an itching
dermatitis reaction to at least one ne perfume or a ershave, and reactions
to other perfumed products. e probable allergy involved reacting to
one or more perfumed products (e.g., deodorant) but no speci c perfume

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being identi ed as causing the clinical reaction. e
possible allergy meant reacting to various cosmetic
products with and without perfume, where materials
other than fragrance constituted the possible cause
of the reaction. Finally, those identi ed without a
fragrance allergy had never reacted to a perfumed
material.
is questionnaire allows the dermatologist to
determine which patients have the highest probability
of fragrance contact allergy before patch testing. With
the proper clinical history, patients then proceed
to the patch testing of FM1 and FM2, to de ne the
possible role of fragrance in the dermatitis.
Figure 1. Typical allergic contact

derm atitis of the axilla due to a deodorant
fragranc e Note: The fold of the axilla
is partly spared; a c lassic observation
believed to be due to axillary sw eating
diluting the allergen.
As IFRA has learned

m o re ab o ut frag ranc es,
rec o m m end ed
c o nc entratio ns fo r
frag ranc e use c o ntinue
to d ec rease.
Pa t c h Te s t ing
Patch testing is used by dermatologists to
determine if a chemical is causing an individual's
allergic in ammatory reaction on the skin. Usually, a
sampling of diluted potential allergens is arranged in
a grid pattern on an individual's back. Since this test
is for a type IV hypersensitivity reaction, or delayed
type of hypersensitivity, the skin must be checked
two and four days later to assess for reaction, which is
contained to the area of application.
Testing with FM1 is a common practice when
dermatitis is suspected to be from fragrance. Nardelli
et al.3 showed that 9.6% of patients investigated
for fragrance contact allergy reacted positively to
FM1, and 6% to FM2. Of those with suspected
fragrance induced dermatitis, sensitivity for FM1 has
been shown to be 27.2%, and 8.7 21.5% for FM2.7
Additional patch testing for FM2 helps to minimize
fragrance-induced clinical dermatitis false negatives, with an estimated additional 6% of patients
identi ed.3
Additionally, Larsen et al. report false negatives
to be 33%, suggesting neither FM1 or FM2 alone
are su cient screening tools.8 Schnuch noted that
46% of patients react negatively when tested for the
individual constituents of FM1 but positively to the
mixture.9 is may be due to false positive reactions
to the mix, lowered allergen threshold, false negative
Photo Credit: Jean-Marie Lachapelle, MD
Figure 2. Positive patch test reaction

scored as ++ ac cord ing to International
Contact Derm atitis Research Group
recom m endations
Photo Credit: Jean-Marie Lachapelle, MD
Vol. 129, No. 4 | May 2014
reactions to individual constituents, or the existence

of a compound allergy.6
To increase sensitivity, Hesiterberg et al. recommend using four screening markers: FM1, FM2,
M. pereirae (balsam of Peru) and HICC. Further, in
addition to the 14 fragrance allergens used in FM1
and FM2 for patch testing, they recommend testing
for 12 fragrance constituents, including: butylphenyl methylpropional, Evernia furfuracea, linalool,
benzyl salicylate, benzyl alcohol, anise alcohol,
benzyl cinnamate, amylcinnamyl alcohol, limonene,
alpha-isomethyl ionone, benzyl benzoate and methyl
2-octynoate.1 Recently, additional allergens have
been commercialized, including hydroperoxides of
linalool, hydroperoxides of limonene and perfume
mix (Mx-08).10
Fr a g r a n c e C o nc e n t r a t io n
Consideration of fragrance concentration and its
impact on dermatitis adds another layer of complexity when managing fragrance-allergic consumers.
e International Fragrance Research Association
(IFRA) publishes safe guidelines on fragrance use.
Based on research, the IFRA Code of Practice
includes 186 standards that either restrict or prohibit
the use of selected fragrance materials for all types
Vol. 129, No. 4 | May 2014
of applications. Producers of fragranced cosmetics

and household products are expected to comply with
IFRA standards.11
In addition to regulating speci c fragrance
compounds, IFRA sets acceptable fragrance exposure
levels. Allowable concentrations are determined by
the quantitative risk assessment of factors such as
volume of use, dermal exposure, and structural alerts
for dermal sensitization. Broad categories organize
products by exposure level, including: Category 1
for lip products, toys and waxes for mechanical hair
removal; Category 2 for deodorants, antiperspirants
M arket Intelligence
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CFBVUZQSPEVDUTI BW
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SBHSBODF
DPNQPOFOUPG
UFOI BW
FBOFEHFJOUIFNBSLFUQM
BDF
BOEUIBURVBM
JUZG
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| 35
Res ear ch | C&T

and fragranced bracelets; and Category 3 for a ershave, eye products, facial creams, tampons, baby
creams and body paint for children. Exposure levels
vary by product, but the concentration level for
eugenol, for example, is 0.20% for Categories 1 and 2
and 0.50% for Categories 3 (and 4).11
As IFRA has learned more about fragrances, recommended concentrations for fragrance use continue
to decrease. is suggests fragrance concentration is
relevant to dermatitis; however the tolerated level is
unlikely to be discerned by patch testing alone.
0.05% and 0.005% w/v of eugenol, a weak sensitizer.

e highest ROAT concentration (0.5%) was selected
based on the current allowable concentration for use
in hydroalcoholic products according to the IFRA.
ese tests involve the once or twice daily application
of the product to a convenient anatomic site, such as
the elbow pit, for up to 28 days. At the levels mentioned, eugenol did not induce reactions in eugenol
patch test positive volunteers.12 Additional use test
studies with fragrances are welcomed by the dermatology and dermatotoxicology community.
Re p e a t O p e n A p p lic a t io n Te s t
D e r m a t o lo g is t
Re c o m m e n d a t io n s
IFRA regulations underscore the importance of

fragrance concentration, and an understanding of
time-dose relationship is important for consumers
to integrate into their daily use or abstinence from
scented products. Svedman et al. described threeweek, repeat open application tests (ROAT) in which
individuals were tested with concentrations of 0.5%,
Table 1. Fragrance M ix Allergens and

Additional EU Allergens
Material
Fragrance mix
Amyl cinnamal
FM1
Cinnamal
FM1
Cinnamyl alcohol
FM1
Citral
FM2
Citronellol
FM2
Coumarin
FM2
Eugenol
FM1
Evernia prunastri
FM1
Farnesol
FM2
Geraniol
FM1
Hexyl cinnamal
FM2
HICC
FM2
Hydroxycitronella
FM1
Isoeugenol
FM1
Additional EU Allergens
Alpha-isomethyl
ionone
Butylphenyl
methylpropional
Amylcinnamyl alcohol
Evernia furfuracea
Anise alcohol
Limonene
Benzyl alcohol
Linalool
Benzyl benzoate
Methyl 2-octynoate
Benzyl cinnamate
Sorbitan
sesquioleate
Benzyl salicylate
If a patient's clinical history suggests contact

dermatitis from a fragrance, the fragranced product should be discontinued for 8-12 weeks. If this
improves the dermatitis, patch testing for common
aromatic allergens included in FM1 and FM2
should be attempted to identify clinically relevant
fragrances. Dermatologist o ces equipped with more
complete fragrance batteries permit the identi cation of additional fragrance allergens. In Europe, if
the concentration of 26 known fragrance allergens
exceeds 100 ppm in a rinse-o product, or 10 ppm in
a leave-on skin products, they must be listed on the
label so the consumer has more freedom and awareness when choosing products.13
If an individual tests positive to FM1 or FM2,
some dermatologists advise that individual to avoid
all FM1 and FM2 fragrance compounds. is is not
straightforward for that person, as these ingredients
are not always listed on products. Other dermatologists advise patients to eliminate exposure to all
fragrances. is is not a simple task, considering the
pervasive use of fragrance in products. It may be
possible to nd a fragrance-free body lotion, but few
brands produce entire fragrance-free product lines.
If an individual is motivated to use a speci c
product a er the period of abstinence, a ROAT can
be performed to ascertain if there is an acceptable
exposure concentration. As noted, this consists of
once to twice-daily applications of the product for up
to 28 days, typically to the elbow pit. Identi cation of
the o ending fragrance and a ROAT to understand
appropriate concentration may allow the patient
to resume use of the fragranced product in a safe
manner.
Much remains to be learned about the clinical
relevance of a positive fragrance mix patch test.
Banning life-long fragrance use is practiced but o en
cumbersome for a consumer. With the exception of
overt dermatitis from ne fragrances and toilet water,
and axillary dermatitis from deodorants and antiperspirants, clinical relevance is o en not obvious.
Vol. 129, No. 4 | May 2014
References
1. MV Heisterberg, T Menn and JD Johansen, Contact allergy to the 26 speci c fragrance ingredients
to be declared on cosmetic products in accordance with the EU cosmetics directive, Contact Dermatitis 65(5) 266 275 (2011)
2. MV Heisterberg et al, Deodorants are the leading cause of allergic contact dermatitis to fragrance
ingredients, Contact Dermatitis 64(5) 258 264 (2011)
3. A Nardelli, A Carbonez, J Drieghe and A Goossens, Results of patch testing with fragrance mix 1,
fragrance mix 2, and their ingredients, and Myroxylon pereirae and colophonium, over a 21-year
period, Contact Dermatitis 68(5) 307 313 (2013)
4. WG Larsen, Allergic contact dermatitis to the perfume in Mycolog cream, J Am Acad Dermatol 1(2)
131-133 (1979)
5. PJ Frosch, JD Johansen, IR White and JC Congress, Fragrances: Bene cial and Adverse Effects,
Springer, New York, USA (1998)
6. L Schollhammer, KE Andersen and CG Mortz, The diagnostic value of patch tests with two fragrance
mix I preparations for detection of clinically relevant perfume allergy, Contact Dermatitis 66(6) 350 352
(2012)
7. PJ Frosch et al, Patch testing with a new fragrance mix detects additional patients sensitive to
perfumes and missed by the current fragrance mix, Contact Dermatitis 52(4) 207 215 (2005)
8. W Larsen et al, A study of new fragrance mixtures, Am J Contact Dermatol 9(4) 202 206 (1998)
9. A Schnuch, J Geier, W Uter and PJ Frosch, Another look at allergies to fragrances: Frequencies of
sensitisation to the fragrance mix and its constituents, Exog Dermatol 1(5) 231 237 (2002)
10. Patch Test Products and Reference Manual 2014, Chemotechnique Diagnostics, www.chemotechnique.se/ck nder/user les/ les/Patch%20Test%20Products%20and%20Reference%20Manual%20
2014%20-%20For%20digital%20distribution(1).pdf (accessed Mar 14, 2014)
11. IFRA RIFM QRA Information Booklet Version 6.0, International Fragrance Association, www.ifraorg.
org (2011) (accessed Oct 15, 2013)
12. C Svedman et al, Does the new standard for eugenol designed to protect against contact sensitization protect those sensitized from elicitation of the reaction? Dermat Contact Atopic Occup Drug
23(1) 32 38 (2012)
13. DA Buckley, Fragrance ingredient labelling in products on sale in the UK, Br J Dermatol 157(2)
295 300 (2007)
GU ES T CON TRIB UTORS

Garrett Coman is a senior medical student at the University of Utah,
currently completing a research fellowship at Maibach's lab. He has a
background in economics and biomedical engineering, with a focus on
medical innovation and research in dermatology.
Nick Blickensta is a senior medical student at the University of Utah, with

a background in psychology and biomedical engineering. He is currently
completing a research fellowship at Maibach's lab, performing research
focused on dermatopharmacology and dermatotoxicology.
Ashley Edwards is a senior medical student at Touro University California.

She plans to pursue pediatric dermatology. A er graduating from Columbia
University in chemical engineering and visual arts, she worked in medical
ethics and public health research at Columbia University Medical Center.
Vol. 129, No. 4 | May 2014
| 37
Res ear ch | C&T
M olecular Biology in
Future Skin and Hair Care

Howard Epstein, PhD
EMD Chemicals, Philadelphia, USA
KEY WORDS
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ABSTRACT
Techniques developed
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biology are currently
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cosmeceutical ingredients
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and hair health and
appearance. Several
relevant developments are
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Save to
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new Web tool.
olecular cell biology is a eld that investigates how cells develop,

operate, communicate and control their activities. Cells communicate with body tissue, which is composed of cells. Tissue
communicates with other tissues and organs throughout the body. Generally the transfer of information between cells, tissues and organs is through
the interaction of proteins.
Techniques developed in the eld of molecular biology are currently
being used to screen cosmeceutical ingredients for skin and hair care
applications. New ndings are published on a daily basis, providing insight
with respect to future innovations for skin and hair health and appearance.
Several relevant developments are reviewed here.
G e n e Ex p r e s s io n a nd Ep ig e n e t ic s
Gene expression is the process by which information from a gene is
used to direct the synthesis of a functional gene product most frequently,
a protein. Gene expression involves a large number of individual genes, and
is a complex activity with many layers of control. It is a critical component
of normal growth and development, and disruption or changes in gene
expression are responsible for many diseases, as well as aging. UV radiation, chemical exposure, viruses, chronic in ammation and oxidative stress
are all associated with skin conditions and aging. Recent publications also
suggest that impaired cellular energy metabolism has an impact on the
condition of skin.1, 2
Epigenetics is the study of heritable changes in gene expression or
cellular appearance caused by mechanisms other than changes in the underlying DNA sequence. Another way to describe epigenetics is the branch of
molecular biology related to mechanisms that a ect gene activity continually occurring inside the cells of the body. Epigenetic events do not alter the
DNA of the cell, rather they change the shape and chemical behavior of the
molecules that form DNA. ese changes alter the instructions coded in the
DNA. Changes in gene expression may be temporary or they may be passed
down to the next generation. Epigenetic e ects can by stimulated by the
environment, lifestyle choices or what are currently perceived as random
events, which commonly occur when DNA is being formed mutations, for
example.

Vol. 129, No. 4 | May 2014
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H a llm a r k s o f A g in g
3
A recent review of the hallmarks of aging noted

nine common denominators that appeared most
frequently in the scienti c literature. ese are: 1)
genomic instability, i.e., damaged genes; 2) telomere
attrition or exhaustion, whereby cells no longer
proliferate; 3) epigenetic alterations, as described
above; 4) loss of proteostasis, meaning impaired cell
signaling caused by dysfunctional protein signaling;
5) deregulated nutrient sensing, examples of which
The p ro m ise o f g ene

exp ressio n and
ep ig enetic stud ies is to
id entify targ et p ro d uc ts
to im p ro ve hum an
health d uring ag ing .
are the insulin pathway or tumor suppressors; 6)
mitochondrial oxidative metabolism, e.g., white/
brown fat created and stored in the body, or mitochondrial dysfunction when DNA repair is hindered;
7) cellular senescence when cells are not replicating
properly; 8) stem cell exhaustion, which is o en
observed in muscle tissue and epidermis this is
a state of reduced cell division and proliferation or
reduced cell regeneration; and 9) altered intercel-
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lular communication, especially seen when cells are

subject to increased in ammation and a weakened
immune system. A tenth hallmark of aging would
be low grade chronic in ammation, referred to as
in ammaging.
e authors of the hallmark publication note that
a major challenge is dissecting the interconnectedness
of these denominators with their relative contributions to aging.3 e ultimate goal is to identify target
products to improve human health during aging, and
this is the promise of gene expression studies including epigenetics-related research.
M a r k e t ing M e c h a n is m s
Historically, the cosmetic chemist formulated
a product and the marketing department created
a story with limited scienti c documentation to
promote it. is paradigm has shi ed to focus
marketing on the proposed biological mechanism(s)
underlying skin concerns the product is intended to
address such as acne, dry skin, in ammation and
aging. Today, it is also expected that cosmetic ingredient suppliers provide a data package that includes a
mechanism of activity with supporting laboratory test
data, o en in vitro.
For example, the ingredient vitamin B3, or
niacinamide, could be marketed based on lab studies
showing its ability, in co-cultures of melanocytes and
keratinocytes, to improve skin hyperpigmentation by
inhibiting the transfer of pigment-bearing melanosomes from melanocytes to keratinocytes.4 Also,
certain soy products are reported to improve the
evenness of skin tone by modulating the activity of a
protease-activated receptor-2 (PAR-2) found in skin.5
Peptides function as cell signaling molecules and
enzyme inhibitors, and these mechanisms have been
identi ed using molecular biological techniques.6
And the current opinion, based on epigenetic studies,
is that while the cell signaling role of antioxidants
is important, this activity may have greater power
unrelated to antioxidant activities.7, 8
N r f2 Tr a n s c r ip t io n Fa c t o r
A transcription factor is a protein that binds to
speci c DNA sequences to control the ow of genetic
information, i.e., transcription, from DNA to messenger RNA. Transcription factors may work alone
or with other proteins in a complex. e Nuclear
Factor E2-Related Factor 2 (Nrf2) signaling pathway
(see Figure 1) is one area of current interest. Nrf2 is
a transcription factor that activates more than 200
genes crucial in the metabolism of drugs and toxins,
signaling for protection against oxidative stress and
in ammation. It is a stress-sensing genetic transcription factor that is thought to be a master regulator of
cellular responses to oxidative damage. Nrf2 is one
Vol. 129, No. 4 | May 2014
of a variety of proteins that controls which genes are

applied topically to damaged skin can modulate
turned on based on the speci c needs of the cell.
in ammation and reduce scar tissue formation. Heng
Nrf2 plays a crucial role in stabilizing proteins
et al.11 also observed topically applied curcumin
and removing damaged ones from cells. Properly
improved psoriasis a er three to eight weeks of treatment. Interestingly, Sonavane12 obtained equivalent
functioning proteins are essential for cellular communication, and Nrf2 was recently identi ed as a
results with topical and dietary curcumin for the
signaling mediator connected with the e ects of
treatment of skin cancer.
caloric restriction through cross-talk with other metNrf2 recently was also found to be important in
abolic signaling pathways. For example, it interacts
wound healing and to skin exposed to UVB radiawith p53 and NF- b, and this combined interaction
tion. Higher levels of Nrf2 protein expression were
is thought to be a guardian of life span and to protect
found in the upper layer of skin than in the lower
against age-related disease.
It was also shown that
signaling between Nrf2 and
Figure 1. The m echanism of Nrf2; Nrf2 is in the cytoplasm o f
cellular metabolic pathways
cells and attached to the protein Keap 1. When oxidative stress
is associated with insulin
reac hes the cell, Nrf2 is released from Keap 1 and translocates
signaling. In this way, it is
to the cell nuc leus. In the nucleus, Nrf2 bind s to another protein,
hypothesized that Nrf2 sigelec trophile- response elem ent (ARE), in the prom oter region of the
naling is a mediator of the
nucleus (sho w n as a dark rec tangle). In this w ay, a variety of genes
e ects of caloric restriction,
are activated through transc ription. Source: o penw etw are.org /
including longevity.
w iki/ 20.109_M OD3_Researc h_Proposal
Co ee, chocolate,
turmeric, olive oil, broccoli, garlic, green tea and
blueberries contain chemical components generically
called phytochemicals,
and many of these have
been reported to induce
the expression of enzymes
in uencing cellular antioxidant defense mechanisms.
Multiple biological pathways are involved; most
frequently, those for Nrf2/
keap1, NF-kB, sirtuinFOXO are reported.9
N r f2 Pa t h w a ys
a n d Sk in
In the majority of
research papers published
on the therapeutic activity
and e cacy of synthetic
and natural products,
the test materials were
ingested by or injected
into test subjects or animal
models. However, a select
number of studies have
been published evaluating
the e cacy of products
applied topically. Heng,10
for example, published data
indicating that curcumin
Vol. 129, No. 4 | May 2014
Table 1. Natural M aterials With Nrf2 Activity and Their Sourc es

Material
Source
Curcumin
Tumeric
Carnosol
Rosemary
Quercetin
Dark colored vegetables, onions
Resveratrol
Red wine
Perillaldehyde
Cherries
Isothiocyanates sulforaphane
Broccoli and cabbage
Catechins
Green tea
| 41
Res ear ch | C&T
Table 2. Com m on Phytochem icals and Their M o des of Action 17

Mode of action
Example phytochemicals
Filtering light
Alkaloids as in caffeine; avonoids in red wine, cocoa and

tea; epigallocatechin 3-gallate in green tea
Inhibiting chronic in ammation
Epigallocatechin 3-galllate
Modulating immunosuppression
Epigallocatechin 3-galllate
Inducing apoptosis
Scavenging reactive oxygen species (direct
antioxidant)
Carotenoids
Activating antioxidant pathways

(indirect antioxidant)
Isothiocyanate sulforaphane in broccoli
layer, and lower layer skin cells die at a higher rate

than upper layers when exposed to UVB probably
for this reason.13 Another study found quercetin
protected keratinocytes against UVA exposure in cell
cultures via Nrf2.14 Table 1 lists the natural ingredients that have been shown to provide skin bene ts
via the Nrf2 pathway; Table 2 lists the di erent ways
phytochemicals provide bene ts for health.15
Sk in Lip id s a n d
Po s s ib le Ro le o f N r f2
Skin is the largest organ of the body. e innermost layer is a subcutaneous fat layer. Next is the
dermis containing broblasts that produce collagen
and elastic bers. Within the dermis are specialized organelles including sebaceous glands, sweat
glands and hair follicles. Nerves and blood vessels

are localized in the dermis as well. e epidermis is
the outermost layer of skin, where highly active lipid
synthesis occurs.
Studies in lipid metabolism have not previously
been of major importance to skin care product development. However, it is becoming an area of greater
interest as studies in molecular genetics increasingly
show that barrier permeability abnormalities are the
primary cause of atopic dermatitis and other skin
diseases. Further, the sebaceous glands play a key role
in acne, dry skin and other skin conditions. In short,
there is renewed interest in lipid metabolism and elucidating its impact on skin.16 (For more on this topic,
see the article by Dayan and Halperin on Page 22.)
Nrf2 regulates lipid metabolism in the liver and
Vol. 129, No. 4 | May 2014
adipose tissue, although the role of Nrf2 with respect

to regulating adipose and other skin conditions is not
clear at this time. Ceramide, cholesterol and phosphatidic acid are the basic structures of cell membrane
lipids, and they can be modi ed by energizing cells
with glucose, which facilitates metabolism. Ceramides are the major component of the stratum corneum
and essential for a functioning permeability barrier;
the role of Nrf2 in regulating lipid barrier function is
an area of current investigation.17
Skin surface lipids are a mixture of sebum and
keratinocyte membrane lipids.18 Major lipid components of sebum include squalene, wax esters and
triglycerides. e squalene found in skin is di erent
from squalene in other body parts. In humans, about
60% of dietary squalene is absorbed and transported
in serum to tissue by very low density lipoproteins.
e greatest accumulation in skin is from sebocyte
concentrations. In the liver, it is metabolized to
squalene epoxide and converted to lanosterol. In the
sebaceous gland, two key oxygen-regulated enzymes
are involved in squalene metabolism: squalene
synthase and squalene oxidocyclase. is is biologically important, as peroxidable squalene is known to
be a key mediator of skin reactions to environmental
stressors.19 Previously, little research on epidermal
lipids was available but more recently, the biological
signi cance of sebum lipids to skin and hair has
become the subject of research activity; speci cally
for acne, seborrheic dermatitis, pityriasis versicolor
and androgenic alopecia.18
In relation, long-wavelength UV radiation
(UVA-1, 340-400 nm) causes oxidative stress to skin
cells, to which cells respond by producing detoxifying enzymes and antioxidants. e e ects of UVA-1
exposure combined with oxidized lipid treatments
on human dermal broblasts and keratinocytes have
been studied. An increase in the accumulation of
nuclear DNA binding of Nrf2 was observed. UV
exposure to skin may result in the oxidation of lipids
found in the membrane layer surrounding skin cells.
e signi cance of the DNA binding of Nrf2 is that
Nrf2 expression will be silenced, resulting in the
absence of antioxidant defense mechanism activation.
In the referenced paper,20 investigators view the lipids
in skin as speci c signaling mediators, whereas previously UV-generated lipid products were considered
to be merely the end result of UV exposure.20
e quality of skin surface sebum lipids, including sebum oxidant levels, and the transport of these
products to skin is an area requiring further research.
e external lipid lm represents a reliable in vivo
biomarker that has the potential to indicate degrees
of environmental stress, to evaluate drug delivery
through the skin and chemical reactions on the
Vol. 129, No. 4 | May 2014
skin, and to assess cross reactions of jewelry, textiles,

cosmetics, drugs, industrial chemicals and other
materials that come into contact with skin.18
C o n c lu d in g Re m a r k s
e Fitzpatrick scale for skin types was developed
about 40 years ago, is based on skin's complexion
and appearance, and provides subjective reporting of
an individual's reaction to UV exposure. Since that
time, molecular biology has advanced considerably.
Currently, the most common methods to collect
data include genome sequencing, the study of single
nucleotide polymorphisms, epigenomics and the
evaluation of many di erent types of RNA expression in cells. As the cost and time to conduct such
studies decreases, an increase in published research
will provide new data and formulation opportunities
for skin and hair care product development. e
information generated from these studies is daunting,
and there is much to be interpreted from it. Further,
the analysis and interpretation remains an inexact
science.21 However, one can envision the day, perhaps
within the next 10 years, when personalized medicine
and skin and hair care products will be common.
| 43
Res ear ch | C&T

References
1. TN Seyfries and LM Shelton, Cancer as a metabolic disease,
Nutrition Metabol 7(7) 1-22 (2010)
2. RK Singh, A Sudhakar and BL Lokeshwar, From normal cells to
malignancy: Distinct role of pro-in ammatory factors and cellular
redox mechanisms, J Cancer Sci Therapy 3(4) 70-75 (2011)
3. C Lopez-Otin, MA Blasco, L Partridge, M Serrano and G
Kromer, The hallmarks of aging, Cell 6 1194-1217 (2013)
4. A Greatens et al, Effective inhibition of melanosome transfer
to keratinocytes by lectins and niacinamide is reversible, Exp
Dermatol 14 498-508 (2005)
5. ER Sharlow et al, The protease-activated receptor-2 upregulates
keratinocytes phagocytosis, J Cell Sci 113 3093-3101 (2000)
6. L Zhang and TJ Falla, Cosmeceuticals and peptides, Clin
Dermatol 27 485-494 (2009)
12. K Sonavane et al, Topical curcumin-based cream is equivalent

to dietary curcumin in a skin cancer model, J Skin Canc 1-9,
Doi:10.1155/2012/147863 (2012)
13. M Schafer et al, Nrf2 establishes a glutathione-mediated gradient of UVB cytoprotection in the epidermis, Genes and Devel 24
1045-1058 (2010)
14. S Kimura, E Warabi, T Yanagawa, D Ma, K Itoh and Y Ishii,
Essential role of Nrf2 in keratinocyte protection from UVA by
quercetin, Biochem and Biophys Res Comm 378(1) 109-14
(2009)
15. A Dinkova-Kostova, Phytochemicals as protectors against
ultraviolet radiation: Versatility of effects and mechanisms, Planta
Med 74 1548-1559 (2008)
16. KR Feingold KR, The importance of lipids in cutaneous function,
J Lipid Res 48 2529-30 (2007)
7. T Finkel and NJ Holbrook, Oxidants, oxidative stress and the

biology of aging, Nature 408(9) 239-7 (2000)
17. Y Ishibashi, A Kohyama-Koganeya and Y Hirabayashi, New

insights on glycosylated lipids: Metabolism and functions,
Biochem et Biophysica Acta 1831 1475-1485 (2013)
8. S Oter, S Jin, L Cucullo and HJ Damien Dorman, Oxidants and

antioxidants: Friends or foes? Oxid Antioxid Med Sci 1(1) 1-4
(2012)
18. C De Luca and G Valacchi, Surface lipids as multifunctional

mediators of skin responses to environmental stimuli, Mediator
In amm 1-11 (2010)
9. A Speciale, J Chira si, A Saija and F Cimino, Nutritional antioxidants and adaptive cell responses: An update, Curr Mol Med
11(9) 770-89 (2011)
19. Y Yamamoto, Role of active oxygen species and antioxidants in

photoaging, J Dermat Sci 27(1) s1-s4 (2001)
10. MC Heng, Curcumin targeted signaling pathways basis for antiphotoaging and anti-carcinogenic therapy, Int Soc Dermatol 49
608-622 (2010)
11. MC Heng, MK Song, J Harker and MK Heng, Drug-induced
suppression of phosphorylase kinase activity correlates with
resolution of psoriasis as assessed by clinical, histological and
immunohistochemical parameters, Br J Dermatol 143 937-49
(2000)
20. F Gruber et al, NF-E2-related factor 2 regulates the stress

response to UVA-1-oxidized phospoholipids in skin cells, FASEB
J 24(1) 39-48 (2010)
21. F Bont , 11th annual LVMH recherch symposium: Skin rejuvenation, Eur J Dermatol 22(3) 432-6 (May/Jun 2012)
Vol. 129, No. 4 | May 2014
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Res ear ch | C&T
Review and M odern Advances of
Retinoids for Cosmetics

Steven Isaacman, PhD, and
Michael Isaacman, PhD
Nanometics LLC, Great Neck, NY, USA
Peter Smith
New York University, New York City, USA
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he vitamin A metabolite retinol is essential for life, and has been shown
to exhibit a diverse range of biological functions. Natural as well as synthetic molecules that are structurally related to vitamin A are referred
to as retinoids, and typically consist of a polyene chain linking a cyclic end
group to a polar end group (see Figure 1). Within the cell, retinoids function as signaling molecules, and play important roles in vision, embryonic
development, cell proliferation, cell di erentiation and immune functions.1
Vitamin A is stored intracellularly as retinyl esters, which a er conversion to
retinol, are oxidized to more bioactive retinaldehydes or retinoic acids. e
structural abundance of retinoids (see Figure 2) and subsequent bioactivity
allow for a variety of therapeutic applications within the pharmaceutical and
cosmetic space.
C h e m o t h e r a p y a nd C a n c e r Pr e v e n t io n
In carcinogenesis, the intracellular levels of retinyl esters are greatly
reduced, compromising retinoid signaling.2 In relation, extensive work has
been conducted to examine the e cacy of retinoids to restore signaling for
cancer therapy. ese studies show retinoids are both chemopreventive and
chemotherapeutic, with their activity attributed to abilities to induce cell differentiation, arrest cell proliferation and promote apoptosis in cancer cells.3
All-trans retinoic acid, or tretinoin, is the most extensively studied
Figure 1. Chem ic al structure o f vitam in A, retinol, show ing

the c haracteristic s o f retinoids; cyclic end group (green),
polyene linker (black), and polar end group (blue)
Save to
My Li brary, a
new Web tool.

Vol. 129, No. 4 | May 2014
n
o
i
t
a
c
i
n
u
m
m
o
c
l
l
ce
Communicating, exchanging messages or sharing data are today

naturally anchored in our everyday life. This hyper-connection also exists
at the skin level through biological networks between cells and tissues
but can be altered by internal or external stresses.
Acting as modulators of endogenous systems, SILAB's actives allow the
skin to react and adapt to its environment for maintaining an optimal
homeostasis.
MESSAGE MODULATORS
UNFLAMAGYL
Yeast biopeptides
VOLUNAGE
Peony oligosaccharides
EPIDERM IS
DERMIS
Anti-inflammaging
DERMIS
HYPODERMIS
Replumping
TRANSPORT CONTROLLERS
WHITONYL
Algae oligosaccharides
PRO-LIPISKIN
Yeast mannans
MELANOSOM ES
Depigmenting
LIPIDS
Restructuring & moistur izing
RECEPTION SYSTEM REGULATORS
NACHYLINE
Yeast peptides
VEDERINE
Chicory oligofructosans
nACh RECEPTOR
Restructuring & barrier function
VITAMIN D RECEPTOR
Restructuring & repairing
Res ear ch | C&T

retinoid for cancer therapies, and has been evaluated
in clinical trials for the treatment of lymphoma, melanoma, lung cancer, neuroblastoma and leukemia.3
Retinoids have shown e cacy at preventing the onset
and development of cancer, especially precancerous
skin lesions. UV irradiation can cause a de ciency
of vitamin A; as such, retinoid treatment can prevent
aberrant signaling and skin cancer development.3
A n t i- A g ing Pr o p e r t ie s
Intrinsic skin aging is a natural process, although
the majority of changes in skin appearance and health
are attributed to sun exposure.4 Natural skin aging is
mainly characterized by ne wrinkling and sagging,
whereas photoaging causes increased pigmentation,
deep wrinkling, sallowness and dryness. Further differences between intrinsic aging and photoaging can
be seen at the cellular level. Intrinsically aged skin is
thinner, with lower levels of collagen and broblasts.
Photoaged skin is thicker, with the accumulation of
elastin and disorganization of collagen.5
Retinoids can be e ective at repairing photodamaged skin by repairing or halting the degradation
of collagen, elastin and hyaluronic acid, the main
structural constituents of skin. All-trans retinoic
acid is commonly prescribed to successfully improve
the appearance of aged skin as well as prevent the

unfavorable e ects of aging. is is accomplished by
stimulating dermal collagen production, reducing
collagen degradation, and promoting epidermal turnover.6, 7 Non-prescription retinoids such as retinol,
retinaldehyde and retinyl propionate also e ectively
lessen skin wrinkling by stimulating extracellular
collagen production.8 In the skin, these derivatives
are converted to the more active retinoic acid,9 which
results in similar e ects to retinoic acid.10
Retinoid e cacy is backed by numerous clinical
trials, and thus remains the gold standard for the
treatment of skin aging.11
Effe c t iv e A c n e Tr e a t m e nt
e use of retinoids to improve acne has been

successful as well.12 Here, retinoids function by
exhibiting immunomodulatory and anti-in ammatory responses, in addition to stimulating collagen
production and increasing epithelial turnover.13 ese
activities also treat acne scarring.14
All-trans retinoic acid has repeatedly been proven
to signi cantly reduce mild to moderate in ammatory acne and blackheads, making it a frequently
prescribed acne medication.13 e most e ective
acne therapy, however, is oral 13-cis-retinoic acid,
isotretinoin, which typically
cures acne when other treatFigure 2. Structures of various retinoid s c urrently utilized in
ments fail.15 Unfortunately, the
the treatm ent and preventio n of certain cancers, skin aging
use of oral retinoids, which is
and ac ne
most commonly isotretinoin,
is limited due to side e ects
such as teratogenicity, caused
by increased levels of retinoic
acid throughout the body.16 e
topical application of retinol,
however, has not been shown to
a ect levels of retinoic acid in
blood plasma, therefore avoiding teratogenic risks17 although
both tretinoin and isotretinoin
treatments have high irritation
potential, and are light and air
sensitive. Newer generations
of synthetic retinoids, including adapalene and tazarotene,
represent molecules that are
beginning to overcome these
n " DDPS
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A N e w Re t in o id
fo r C o s m e t ic s
e pharmaceutical and
cosmetic utility of retinoids is
impressive, but recent work aims
Vol. 129, No. 4 | May 2014
to push the skin care bene ts even further. One common

goal is to avert the skin irritation and dryness commonly
associated with topical retinoid treatments. Structural
alterations to vitamin A such as molecular alterations
to the cyclic group, polar group or linker have enabled
the construction of synthetic retinoids that harness the
desired activities while simultaneously reducing adverse
side e ects.18 Taking inspiration from stored retinyl esters,
novel esteri ed forms of retinoic acid are also being
investigated. Typically, natural retinyl esters, such as retinyl
propionate, are less active than retinol.8 However, new
synthetic esters, such as hydroxypinacolone retinoate, are
proving to be viable alternatives (see Figure 3).
Hydroxypinacolone retinoate is of great interest
because it is capable of interacting with retinoid receptors without rst being converted to retinol or retinoic
acid.19 is allows for the e ective treatment of skin while
eliminating the irritation and dryness caused by acidic
retinoids. Furthermore, topically applied hydroxypinacolone retinoate e ectively di uses into skin, comparable to
Figure 3. Synthetic retinoid derivatives that

exhibit enhanc ed properties
Adapalene
Tazarotene
Hydroxypinacolone Retinoate
Vol. 129, No. 4 | May 2014
| 49
Res ear ch | C&T

topically applied retinoic acid, again without elevating the levels of retinoic acid in blood plasma, thus
eliminating the risk of teratogenicity.19
6. C Grif ths, AN Russman, G Majmudar, RS Singer, TA Hamilton

and JJ Voorhees, Restoration of collagen formation in photodamaged human skin by tretinoin (retinoic acid), New England J
Med 329 530 535 (1993)
C o n c lu s io n
7. GJ Fisher, Z Wang, SC Datta, J Varani, S Kang and JJ Voorhees,

Pathophysiology of premature skin aging induced by ultraviolet
light, New England J Med 337 1419 1429 (1997)
Clearly, retinoids possess a wide spectrum of

functions. For skin cancer, skin aging and acne, they
have proven both preventive and therapeutic. Successful e orts to create more bioavailable vitamin A
derivatives and retinoid formulations have increased
the industry's reliance on them for cosmetic bene ts.
Recent work to further optimize retinoids and reduce
their side e ects for cosmetic applications has yielded
a new generation of promising molecules.
References
1. R Blomhoff, M Green, T Berg and K Norum, Transport and storage of vitamin A, Science 250 399 404 (1990)
2. X-H Tang and LJ Gudas, Retinoids, retinoic acid receptors and
cancer, Annual Review of Pathology: Mechanisms of Disease 6
345 364 (2011)
3. N Bushue and Y-JY Wan, Retinoid pathway and cancer therapeutics, Advanced Drug Delivery Reviews 62 1285 1298 (2010)
4. MCB Hughes, GM Williams, P Baker and AIC Green, Sunscreen
and prevention of skin aging, a randomized trial, Annals of
Internal Medicine 158, 781 (2013)
5. YR Helfrich, DL Sachs and JJ Voorhees, Overview of skin aging
and photoaging, Dermatology Nursing 20 177 184 (2008)
8. ZD Draelos, The latest cosmeceutical approaches for antiaging,

J Cos Derm 6 2 6 (2007)
9. CK Huang and TA Miller, The truth about over-the-counter
topical anti-aging products: A comprehensive review, Aesthetic
Surgery Journal 27 402 412 (2007)
10. J Varani et al, Vitamin A antagonizes decreased cell growth and
elevated collagen-degrading matrix metalloproteinases and
stimulates collagen accumulation in naturally aged human skin
1, J Inves Derm 114 480 486 (2000)
11. S Mukherjee et al, Retinoids in the treatment of skin aging: An
overview of clinical ef cacy and safety, Clinical Interventions in
Aging 1 327 348 (2006)
12. LF Sandoval, JK Hartel and SR Feldman, Current and future
evidence-based acne treatment: A review, Expert Opinion on
Pharmacotherapy 15 173 192 (2014)
13. A Thielitz and H Gollnick, Topical retinoids in Acne vulgaris:
Update on ef cacy and safety, Amer J Clin Derm 9 369 381
(2008)
14. AE Rivera, Acne scarring: A review and current treatment
modalities, J Amer Acad Derm 59 659 676 (2008)
15. A Shalita, The integral role of topical and oral retinoids in the
early treatment of acne, J Eur Acad Derm and Venereol 15
43 49 (2001)
16. FW Rosa, AL Wilk and FO Kelsey, Vitamin A congeners, Teratology 33 355 364 (1986)
17. GJ Nohynek et al, Repeated topical treatment, in contrast to
single oral doses, with Vitamin A-containing preparations does
not affect plasma concentrations of retinol, retinyl esters or
retinoic acids in female subjects of child-bearing age, Toxicology
Letters 163 65 76 (2006)
18. JH Barnard, JC Collings, A Whiting, SA Przyborski and TB
Marder, Synthetic retinoids: Structure activity relationships,
Chemistry A European Journal 15 11430 11442 (2009)
19. J Gormley, Topical hydroxypinacolone retinoate: Skin diffusion,
receptor activity, metabolism and mildness, Grant Industries Inc.
(2008)
GU ES T CON TRIB U TOR

Peter Smith is currently a
fourth-year undergraduate
student at New York University,
where his research is focused
on the design and synthesis of
bioactive peptidomimetics. He
has developed new methods to
utilize these functionalized oligomers as both
antimicrobial agents and inhibitors of crystal
growth. Smith is the recipient of multiple New
York University undergraduate research grants,
and his work has extended into personal care
with research at Nanometics LLC.
Vol. 129, No. 4 | May 2014
Res ear ch | C&T
Patent Picks:
Soft Hair Hold,

Barrier Function and More
KEY WORDS
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Editor's note: Patent Picks are compiled by the editors from publicly available sources and
cover recent patents issued, or applied for, in the cosmetic and personal care industries and
relevant peripheral markets.
So ft , v o lu m izin g , na t u r a l n is h h a ir h o ld
U.S. Patent 8674036; published March 18, 2014; assignee: Kao Corp.
e present invention relates to a hair cosmetic that imparts a so
feeling and natural nish. It is preferably used to provide u and volume
to a hair style, and prevent y-aways, unwanted curls or kinks. Further,
it can maintain a hair style for a long time without being disturbed by
external factors; e.g., combing the hair with ngers, wind or vibrations.
e composition is obtained by incorporating two types of poly(Nacylalkyleneimine)-modi ed organopolysiloxanes having speci c structures
at a speci c ratio, provided in detail in the patent.
Pe p t id ic h yd r o lyza t e t o r e in fo r c e b a r r ie r
fu n c tio n ing
U.S. Patent 8674072; published: March 18, 2014; assignee: ISP Investments, Inc.
Disclosed in this patent is a peptidic hydrolyzate enriched in bioactive peptide that is capable of reinforcing the skin barrier function and
stimulating epidermal di erentiation. Additionally, a cosmetic and/or
pharmaceutical composition that includes a physiologically acceptable
medium and the peptidic hydrolyzate as active principle are described. e
composition activates the HMG-CoA reductase in the cutaneous cells, in
turn treating the cutaneous signs of aging and photo-aging.
M e la n in s ynt h e s is a n d u s e
U.S. Patent 8673983; published: March 18, 2014; assignee: Loyola University Chicago
Save to
My Li brary, a
new Web tool.
Melanins have been shown to possess a number of interesting pharmacological properties including immunomodulatory activity, photoprotective
activity, metal-binding properties useful as image enhancers in MRI, and
the ability to protect against oxidant-induced tissue damage. According
Vol. 129, No. 4 | May 2014
Res ear ch | C&T

to the inventors, it would therefore be desirable
to synthetically produce melanins in high yields
under reproducible conditions. Further, it would be
desirable to produce novel melanins, especially watersoluble melanins, with antiviral activity. is patent
thus describes the enzymatic and chemical synthesis
of melanins and novel melanins by contacting an
oxidase and a phenolic substrate containing at least
two hydroxyl groups for a time su cient to produce a
melanin. e method can further comprise any of the
following steps: inactivating the oxidase, precipitating
the melanin, and/or purifying the melanin.
Silk d o p e p r o d u c t io n ,
a p p lic a t io n in c o s m e t ic s
U.S. Patent 8674077; published: March 18, 2014;
assignee: Commonwealth Scienti c and Industrial
Research Organization
According to these inventors, silks protein bers
are produced by a wide range of insect and spider
species. Numerous e orts have been made to clone
and express silkworm or spider silks in transgenic
systems but the large sizes and highly repetitive
sequences of these silk genes make them recalcitrant
to expression outside specialized silk glands, and lead
to low protein yields. ere is therefore a need for

methods to produce silk dope from recombinantly
expressed coiled-coil silk proteins.
e present invention relates to methods of
producing silk dope comprising silk proteins with a
coiled-coil structure, such as honeybee silk proteins.
e proteins are obtained from cells producing them,
solubilized by contacting them with a surfactant or an
ionic liquid, and concentrated to produce silk dope.
ey can be used in a variety of area such as textiles,
biomedical products and personal care. Applications
may include cosmetics, skin care, hair care and hair
coloring; in the coating of particles such as pigments;
or: ethoxylation to promote water-oil emulsion
enhancement, siloxylation to provide lipophilic
compatibility, and esteri cation to aid in compatibility with soap and detergent compositions.
D e v ic e , m e th o d fo r b o d y
c o n t o u r in g
assignee: Zeltiq Aesthetics, Inc.
is invention relates to methods and devices
that enable the delivery of radio frequency energy
and cryotherapy applications to adipose tissue for
Vol. 129, No. 4 | May 2014
reducing and contouring body fat. e method can

target a region in the subject at a frequency that
selectively heats brous septae in a subcutaneous
layer of the target region to a maximum temperature
less than a brous septae denaturation temperature.
Furthermore, the method can include removing heat
such that lipid-rich lobules in the subcutaneous layer
are a ected while non-lipid-rich cells and lipid-rich
regions adjacent to the brous septae are not substantially a ected.
Qu a n t ifyin g
e ye la s h e s
b e r s , h a ir s a n d

assignee: Procter & Gamble
Described herein is a method for counting the
number of bers emanating from the surface of a
web substrate. Speci cally, the substrate is assessed by
the parameters of: machine direction, MD; a crossmachine direction, CD; orthogonal and coplanar
thereto; and a Z-direction orthogonal to both said
machine and cross-machine directions. While
intended to assess the so ness of tissues and the like,
this method also may be used for various household,
cosmetic and personal implements, for example,
quantifying facial hairs or eyelashes.
La r g e - a r e a Ra m a n p r o b e
w it h r e d u c e d b a c k g r o un d
uo re sc e nc e
two-dimensional array-collecting anti-Stokes Raman

spectra, and a probe con gured to measure complex
solid samples with reduced background uorescence.
e system collects spectra from an area of 1-mm or
greater, preferably 3-12 mm or more, facilitating the
collection of statistically useful data from nonhomogeneous and laser-sensitive samples. e system
can further be used with lower laser energy density
by expanding the laser to up to 10 mm or more to
analyze skin and the e ects of cosmetics on skin
without causing damage to skin. e probe conforms
to the ANSI standards and is therefore skin safe.
Silic o n e - c o n t a in in g , lm fo r m in g p o lym e r s in n a il p o lis h

a p p lic a t io ns
assignee: Surfatech Corp.
is invention is directed to polymers that are
useful in nail polish applications. Film-forming
polyester polymers are described that are liquid in
solvents such as butyl acetate. ey form a uniform
lm upon application when the solvent evaporates
under atmospheric conditions, yet can be removed
easily and thoroughly by application of a solvent such
as acetone. e polymers of the present invention
having a silicone portion contained therein, which
provides improved exibility, tear-resistance and
superior aesthetics.

assignee: Kaiser Optical Systems
Disclosed in this patent is a compact Raman analysis system combining a near-infrared laser source, a
Vol. 129, No. 4 | May 2014
| 55
Tes t in g | C&T
Proposed M ethod to Evaluate the
Microbiological Stability
of Cosmetics During Use
Nadine Bresciani, Val rie Poulet and
Nathalie Collard
Anemcoli, Lille, France
Rapha l Dugue
Laboratoire Midac, Loos, France
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he aim of antimicrobial e cacy testing (AET) performed during

cosmetic product development is to predict microbial stability and
consumer safety during use. e AET design includes referenced
microbial strains and acceptance criteria. However, such tests do not
include speci c situations resulting from consumer use; for example,
microbial ora encountered in normal environments; repeated insults;
environmental condition variability; the impact of accessories and packaging; and the possibility of localized inoculation, e.g., via caps.
erefore, it was deemed necessary to develop an additional test to
strengthen the investigation. Here, the authors propose an approach to
assess the microbial stability of a product during use, referred to as the
Microbiological Use Test (MUT), and apply this analysis in a few case
studies to predict the microbiological risk of commercial products. e
described test has been used successfully in the development process of
cosmetic products.
Ex p e r im e n t a l D e s ig n
e MUT test assesses, during the product development phase, the
ability of a product to prevent its own microbial contamination during
standard conditions of use. e aim is to perform a quantitative and
qualitative assessment of potential contamination a er a speci ed period of
use. To ensure results, some parameters must be xed; for example, blind
testing, to ensure the product is used under conditions close to reality. Panelists should not be informed of the aim of the study especially that the
product will be microbiologically tested upon return. Also, packaging must
be as close as possible to the nal form, including being comprised of the
same materials and utilizing the same closure system. is is a key point,
as packaging plays an important role during product use and, therefore, in
product contamination.
Timing and conditions also are important. Samples should be returned
directly to the microbiological laboratory without extra manipulation prior
to testing. e time between the last use and the rst test must be xed, e.g.,
72 hr, to limit the recovery of transient microbes but allow for the detection
of the more critical persistent contaminants. Further, it is necessary to have
at least 20 samples involved to ensure a relevant assessment.

Vol. 129, No. 4 | May 2014
Tes t in g | C&T
A general survey should be sent with each product to match the outcome with the user's practices;
also, directions for use, frequency and duration of
application and speci c requirements such as the
use of an applicator should be speci ed. Finally, the
number of subjects included in the study must be
controlled, and they should be realistic potential user
types who are physically located as close as possible
to the laboratory.
De te rm inatio n o f e f c ac y
is b ase d o n p ro d uc t
susc e p tib ility and re d uc e d
c o ntam inatio n.
A n a lys is Pr o to c o l a n d
Ot h e r C o n s id e r a tio n s
Regarding sample analysis, the study must respect
several steps. First, the product contamination
bio-burden should be determined prior to study
execution. Also, some samples should be retained in
the laboratory at room temperature during the study
execution and assayed two or three days prior to the
return of consumer samples for validation; these
results should be lower or equal to the initial count.
Besides MUT standards, the products must meet
the usual microbiological speci cations. Further,
products, identi ed by lot number, require being
assayed for preservative content. Regarding the study
duration, the product must be used as o en as pos-
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(GCImagazine.com
sible; the design proposed here suggests at least three

weeks.
Other considerations involve the design of the
customer survey, directions for use and the study
direction i.e., start date, return date, etc. Product distribution to the selected panel of users and the return
of samples must also be coordinated. As noted, it is
necessary to manage the return date to ensure the delay
between last use and rst testing is within xed limits,
to be able to evaluate a possible regression of microbial
count a er a speci ed time.
Sa m p lin g Pr o c e d u r e s
Testing should include all returned samples using
methods proven suitable, such as ISO 21149, and following usual procedures applied in the laboratories. It
is important to note that microbial contamination due
to product use is typically not homogeneously distributed through the product. erefore, sampling should
occur as close as possible to normal use conditions. e
sampling procedure must include the following steps:
1. No mixing of the product before sampling;
2. Surface sampling for liquid and semi-solid
products;
3. For products in tubes, retaining only the rst
expelled portion; and
4. For products used with an applicator, sampling
must use the applicator, which should be returned
to its original place a er sampling.
A second round of testing is then performed to
assess whether the microbial contamination originally
present was or was not eliminated a er a certain
period. is is a key element of product robustness
assessment. It is recommended that this test be carried
out six days a er the last use; i.e., three days a er the
rst testing, allowing for complete analysis within
one week. is will not be feasible in all instances,
and in certain cases, this could even alter the results
and overall assessment. For example, with powder, a
second sampling performed on a product scraped for
the rst testing will not be relevant since the entire
contamination could have been removed. erefore,
when possible, it is also advisable to scrape only half of
the surface for the rst step, saving the second half for
follow-up tests.
For products packed in tubes, the second dose will
not always be representative, either. erefore, it must
be noted that the absence of recovered contaminant
is not necessarily due to product performance; it may
instead be driven by the absence of a homogeneous
contamination. Finally, to ensure a good understanding
of the results, it is necessary to assess other parameters
such as organoleptic characteristics, physico-chemical
parameters, preservative content and pH.
Vol. 129, No. 4 | May 2014
A s s e s s m e n t Sp e c i c s
Determination of the results is based on two
criteria, described here. e rst is the analysis for
microbial contamination susceptibility; the second is
the analysis for the product's capability to reduce its
original contamination a er a short duration of time.
Susceptibility
rst testing: e rst test is for the
product's resistance to contamination, and is determined by only one criterion: the number of returned
contaminated samples, independent from their levels
of contamination. e presence of 10 cfu/g is enough
to declare a sample as positive. Five levels were
arbitrarily assigned by the authors (see Table 1)
based on the % of positive returned samples. For
example, Level 1 was assigned to the most robust
products.
Reducing contamination second testing:
e second test to reduce contamination is
performed six days a er the last product use.
During this period, products are stored in the
laboratory at room temperature without exposure
to sunlight. In this step, the criteria are di erent
based on product characteristics; i.e., anhydrous
vs. aqueous products.
Aqueous products are usually able to recover
from contamination during the resting phase.
Vol. 129, No. 4 | May 2014
Here, two levels have been set: A) bio-burden reduction, characterized by any signi cant reduction in
count; and B) no bio-burden reduction. Considering
the small number of samples involved (20), and taking into account the size of commercial productions,
products will fail this test as soon as just one sample
fails to reduce bio-burden.
In the case of anhydrous products, three levels
have been set: A) bio-burden reduction, as described
above; B) bio-burden stabilization, an intermediary
level; and C) bio-burden increase, which, even if rare,
must eventually be considered especially for natural
products.
Table 1. Levels of Contam ination

Level of
contamination:
% of contaminated samples:
< 5%
From 5 to 10%
From 11 to 25%
From 26 to 50%
> 50%
| 59
Tes t in g | C&T
M UT A s s e s s m e n t s
e proposed MUT approach will result in
products described as follows. First, each product
will have a resistance level rating from 1 to 5 based
on the rst susceptibility testing. en, based on the
second testing, an additional rating will be assigned:
A or B for aqueous products; A, B or C for anhydrous
products.
e overall assessment will then be rated as:
IP (Insu cient Preservation): For products with
no reduction, or an increase in, bio-burden;
S (Suitable): For products rated at levels between
3 and 5, and with either a bio-burden reduction or no
increase with the second testing; or
RU (Robust during Use): For products rated at
levels from 1 to 2, with either a bio-burden reduction
or no increase as assessed with the second testing.
ese product ratings are further outlined in
Tables 2 and 3.
C o m m e r c ia l Pr o d u c t
Asse ssm e nts
To test the proposed approach, seven commercial products were chosen based on their formula,
function and use or non-use of a preservative system.
e packaging, distribution mode and associated
processes and eventually, certi cation base, i.e., ecolabeling of products were also taken into account.
Speci c choices were made to improve the feasibility
and discriminant power of the MUT model. e
chosen products and their basic information are
included in Table 4.
Challenge tests following NF EN ISO 11930 were
carried out1 on each product except number 7, the
lip gloss. For products 1 through 5, criteria A were
reached for bacteria, yeast and mold. For product 6, a
refreshing lotion, no criteria were achieved; neither A
nor B. e reason for this relates to packaging and is
described later.
Table 2. Aqueous Produc t Assessm ent

First testing
Second testing
1
< 5%
2
5 to 10%
3
11 to 25%
4
26 to 50%
5
> 50 %
A = Reduction
RU
RU
B = No reduction
IP
IP
IP
IP
IP
Table 3. Anhydrous Product Assessm ent

First testing
Second testing
1
< 5%
2
5 to 10%
3
11 to 25%
4
26 to 50%
5
> 50 %
A = Reduction
RU
RU
B = No reduction
RU
RU
C = Increase
IP
IP
IP
IP
IP
Table 4. Selected Products

Product No.
Product Type
Product Information
Foundation
Ecocert certi ed, no listed preservative, 30 mL pump bottle
Foaming gel
Preserved with paraben and potassium sorbate, 200 mL tube
Shower cream
Unpreserved product, 200 mL tube
Face cream
Preserved with phenoxyethanol and benzyl alcohol; presence

of signi cant amount of alcohol based on INCI list positioning
(beginning of the list); 100 mL jar
Face and body cream
Ecocert certi ed, preserved with potassium sorbate and sodium

benzoate, 200 mL jar
Refreshing lotion
Ecocert certi ed, sterilized through UHT process, without

preservative, 150 mL pump bottle
Lip gloss
Anhydrous product, unpreserved with rubber applicator
Vol. 129, No. 4 | May 2014
Results of the MUT assessment are summarized

in Table 5. No contamination was found a er the use
of a face and body cream or a foundation (5). e two
most contaminated products were the foaming gel (2)
and shower cream (3). is is not surprising, as these
are o en stored in areas of risk due to the presence
of water. e preserved shower gel did not provide

better results, which is noteworthy.
For the face cream (4), the ratio of contamination
was high (30%) but concentrations were low (< 10
cfu/g). Nevertheless, this low level of contamination
may be associated with an absence of regression (5%).
Table 5. M ic ro bial Stability During Use

Product
1, Foundation
First Analysis
Second Analysis
Ratio of contaminated
samples
Total of contaminated
product and %
0/20
0/20 (0%)
6/20 (30%) < 10 cfu/g

2, Foaming gel
5/20 (25%) between 10

and 104 cfu/g
na
10/20 (50%) regression
12/20 (60%)
2/20 (10%)
no regression
1/20 (5%) > 104 cfu/g

3/20 (15%) < 10 cfu/g
3, Shower
cream
6/20 (30%) between 10

and 104 cfu/g

11/20 (55%)
3/20 (15%)
no regression
2/20 (10%) > 104 cfu/g

4, Face cream
6/20 (30%) <10 cfu/g
6/20 (30%)
5, Face and
body cream
0/20
0/20 (0%)
na
1/20 (5%)>104 cfu/g
1/20 (5%)
1/20 (5%) no regression
1/20 (5%) < 10 cfu/g
2/20 (10%)
5/20 (25%)
na
6, Refreshing
lotion
7, Lip gloss
1/20 (5%) no regression
1/20 (5%) between 10 and

104 cfu/g
7, Applicator
Vol. 129, No. 4 | May 2014
na
| 61
Tes t in g | C&T
For the refreshing lotion, in only one panelist (10%)
was a high level of contamination observed. In the
absence of preservative system, no regression was
noted.
For anhydrous lip gloss, contaminations are
isolated from the gloss itself as well as from the
applicators. All the contaminants were isolated and
identi ed, and results are summarized in Table 6.
e authors noted a large proportion of cocci coming
from the cutaneous ora, which are directly linked
to use. Also observed was the fact that, among the
recovered microorganisms, only one was speci ed:
Staphylococcus aureus (1/22 Staphylococci recovered).
A large quantity of Gram-positive bacilli were
recovered as well. Another noticeable point was from
the foaming gel (2), where the majority of recovered
microorganisms were Staphylococci, while for the
shower cream (3), Pseudomonas and Enterobacteriaceae were in majority.
Table 7 summarizes the nal product assessments.
Interestingly, none of the tested products were rated
as Suitable (S). e face and body cream, foundation
and lip gloss were classi ed as RU, whereas the face
cream, foaming gel, shower cream and refreshing
lotion were classi ed as IC.
D is c u s s io n : M UT v s . A ET
All products evaluated in accordance with the ISO
document (AET) except the refreshing lotion met the
most stringent criteria. Nevertheless, following the
MUT approach, some products would be classi ed
as IC. is indicates that meeting the AET criteria
is not su cient to ensure a product will not become
contaminated during use.
Also, as noted, the refreshing lotion did not meet
either criteria A or B. In this case, protection of the
ultra high temperature-sterilized formula (UHT) during use was provided by the pack. e protocol showed
5% of samples at most were contaminated (> 104
cfu/g) without any regression observed at the second
analysis. is is the most important demonstration
of the relevancy of the MUT to evaluate the global
microbiological risk during use.
When a formula does not meet either criteria A or
B, or where protection during use is based on packaging, then the MUT is capable of assessing the product's
robustness to prevent contamination. For satisfactory
results (RU and S), this could become acceptable
justi cation for product commercialization. e same
reasoning may be followed for products where AET is
not relevant, i.e., anhydrous lip gloss.
Table 6. Identi c ation o f Contam inants

Product
Total Number of
Isolated Strains
Identi cation and Ratio

Staphylococcus: 9 (45.0%)
2, Foaming gel
20
Micrococcus: 4 (20.0%)
Pseudomonas: 6 (30.0%)
Gram+ rods: 1 (5.0%)
3, Shower cream
14
Enterobacteriaceae: 6 (42.9%)
Aerococcus: 3 (21.4%)
Staphylococcus aureus: 1 (8.3%)
4, Face cream
12
Other Staphylococci: 5 (41.7%)

Gram+ rods: 3 (25.0%)
6, Refreshing lotion
Pseudomonas: 2 (100%)
7, Lip gloss applicator
23
Gram+ rods: 8 (34.8%)
Aerococcus: 1 (4.3%)
Enterococcus: 2 (8.7%)
Vol. 129, No. 4 | May 2014
Table 7. Final Product Assessm ents
Product
% of
Contaminated
Product During
First Testing
Level
( rst analysis)
Product Behavior
(during second
testing)
Conclusion
1, Foundation
na
RU
2, Foaming gel
60
IC
3, Shower cream
55
IC
4, Face cream
30
IC
5, Face and body

cream
na
RU
6, Refreshing
lotion
IC
7, Lip gloss
10
RU
e di erence in results between AET and MUT

can derive from di erent elements. First, during the
MUT assessment, contamination is iterative, whereas
the AET assesses inoculation at one time. e MUT
approach is more appropriate to assess formula
robustness. Further, its contamination is natural
and comes from the user's environment, whereas
Vol. 129, No. 4 | May 2014
the AET relies on simulated contamination with a

limited panel. e likelihood of identifying preservation weakness to some contaminants is therefore
increased using the MUT. Also, packaging is an
important element to prevent microbial contamination during use, and only the MUT can assess this
parameter since the AET only looks at the
| 63
Tes t in g | C&T
formulation independent of the packaging.
ese various elements demonstrate why there is
interest in this test. However, it does have limitations that
must be highlighted. First, the MUT makes sense only if
a second testing can be performed. Also, special attention
must be paid to the sampling and re-sampling conditions
to make sure the data is meaningful. When a product
assessment is based on the second test, the outcome may
be faulty, especially if contamination was diluted despite all
precautionary measures taken during re-sampling. Finally,
knowledge of the nal package is critical to this test, indicating one must know the packaging and have it available
at the time of testing. Since this o en is not the case, this
may delay testing to the latest development phases.
C o n c lu s io n
is study demonstrates, for seven selected products,
the ability of the proposed MUT approach to determine
a product's robustness during use conditions. e specied criteria allows for a standardized investigation of the
microbiological stability of products. Experimental results
demonstrated that products tested three and six days a er
their last use can remain contaminated, as assessed via the
recovery of speci ed microorganisms, potentially altering
their organoleptic properties and product performance.
is risk would not have been identi ed using only the
current AET method, which seems not so predictive, with
the exception of a product failing the test.
MUT appears to be an acceptable alternative to the
AET. It additionally focuses on the element of microbial
stability on the packaging materials required to prevent
microbial ingress. us, microbial risk evaluation of
the global product, i.e., formula plus packaging, during
its normal use in an unprotected environment can be
achieved using the proposed test design. is is of major
interest, especially as the industry is developing more and
more products with limited amounts of or no preservatives. Packaging then becomes a more signi cant part of
a product's preservation, and the MUT is able to assess
packaging performance in situ.
References
1. NF EN ISO 11930, Evaluation of the antimicrobial protection of
a cosmetic product, available for purchase at www.iso.org/iso/
catalogue_detail?csnumber=51037 (Jun 2012)
Vol. 129, No. 4 | May 2014
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June 26 27, 2014

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Phila delp hia, PA
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Tes t in g | C&T
Correlating Aging w ith
Skin's Mechanical and

Optical Properties
Olga Freis, PhD, Gilles Perie and
Andreas Rathjens
BASF Beauty Creations/Care Solutions,
Pulnoy and Essey-l s-Nancy, France
KEY WORDS
CJPNFDI BOJDBMQSPQFSUJFTt
PQUJDBMQSPQFSUJFTt
CSJM
M
JBODFt DPM
PSt
uorescence
ABSTRACT
The aim of this study was
to monitor the evolution
of biomechanical and
optical properties of the
skin with aging. Different
biophysical parameters
were measured, including
skin: elasticity and
rmness, color, brightness,
uorescence emission,
sebum content, hydration
and pH. A signi cant
evolution of the evaluated
parameters with aging was
observed.
Save to
My Li brary, a
new Web tool.
he evolution of skin's biomechanical and optical properties as a function of aging and/or photoaging is one of the main targets of cosmetic
and dermatological research. Many noninvasive devices to measure
skin's biomechanical properties have been developed using alternative
methods such as stretching, torsion, indentation and suction. Measurements of skin deformation a er suction or torsion are the most widely used
techniques in cosmetic research.1, 2
e skin's optical properties play an important role as well, and devices
measuring these characteristics assess re ected light a er illumination
of the skin surface. Di erent noninvasive methods have been proposed
for evaluating skin complexion in vivo. ese include quantitative measurements of skin color, using colorimetry i.e., L*a*b* and Individual
Typological Angle (ITA );3 or of the intensity of specular re ection and the
back-scattering of light from the skin.4, 5 e purpose of this study was to
demonstrate the evolution of the measured parameters with aging, and to
nd the correlation between measured mechanical and optical properties of
the skin.
M e thod s
Test population and body sites: A total of 113 female volunteers ages
18 76 participated in the study. All participants were Caucasian, with no
apparent signs of skin disease. e volunteers applied no cosmetic products
for 24 hr before measurements were taken. Measurements were conducted
on two di erent anatomical regions: the inner forearms and/or the face, i.e.,
the forehead, temples or cheeks.
Biomechanics: e biomechanical properties of the skin were measured
using commercially available standard devicesa, b as well as an internally
developed device, referred to as a corneovacumeter.6 e corneovacumeter measures capacitance between skin and a conductive plate, whereby
capacitance is proportional to skin deformation. With this type of device,
12 simultaneous suctions are realized and the result provided is the mean
of the measurements. While such suction-based devices measure vertical
skin deformation a er suction, torque-based devices measure deformation
a
b
Cutometer SEM 575, Courage & Khazaka

Dermal Torquemeter, DiaStron

Vol. 129, No. 4 | May 2014
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Tes t in g | C&T
a er torsion. For the present study, both standard and
surface methods were used for calculating biomechanical skin parameters.7
Skin color, brightness: Skin color was measured
by colorimeterc in the L*a*b* colorimetric system.
In addition, skin brightness was evaluated by
another internally developed device, referred to
as a brillanometer. is device enables the in vivo
determination of both specular and di use light
re ection, continuously and in numerous directions,
The c o rrelatio n
o f b io m ec hanic al
p ro p erties w ith ag e
w as m o re sig ni c ant
in m easurem ents o f
the fo rearm s than the
fac e, w here p ho to ag ing
inc reased variab ility.
in a contactless manner.8 e studied parameters
were: peak height in specular re ection, peak width
of specular re ection in the middle height, their ratio,
and peak height in di use re ection.
Fluorescence: To acquire uorescence spectra of
the two uorophores present in skin, a spectro uorimeter d was used; the uorophores of interest were:
c
d
CR 200, Minolta
LS 55 Spectro uorimeter, Perkin Elmer
According to the NPD Group:
The U.S. prestige skin care market grew 3%
in the 12 months ending January 2014, reaching
$3.6 billion in sales. Products for the face brought
in $2.8 billion, representing 78% of the business.
The anti-aging crusade has evolved. It's no

longer just about line correction, rming and
wrinkle repair, the emphasis is on things like
radiance, resiliency, texture, evenness and clarity.
n
tryptophan (exc. 295 nm), which re ects epidermal

proliferation; and collagenase digestible cross-links
(exc. 370 nm), which re ect the accumulation of
Advanced Glycation End-products (AGEs) in the
dermis.9
Additional parameters, analysis: Auxiliary
parameters, such as skin pH, sebum levels and hydration state, were measured using a specialized meter e.
e data was analyzed using descriptive statistics and
correlation so ware, and the correlation between
the di erent methods also was evaluated. is
approach allowed the authors to not only determine
the relation of each tested property with aging, but
also determine which properties were statistically
dependent and correlated.
B io m e c h a nic a l Re s u lts
e obtained skin deformation curves were
similar for all devices (see Figures 1 3), and on the
basis of the deformation curves, the biomechanical
parameters were calculated. Here, the authors present
only the ratio parameters that are independent of
skin thickness, i.e.: skin elasticity (ratio Ur/Ue =
[R5]), skin visco-elasticity (ratio Uv/Ue = [R6]) and
skin rmness (Ratio Ur/Uf = [R7]). e correlation of
biomechanical properties with age was more signi cant on the measurements taken on the forearms in
comparison with the face, where the combination
of intrinsic and photoaging increased the variability
of the measurements. is was observed with all
devices used to measure biomechanical properties.
In addition, a strong correlation was found between
the expression of biomechanical parameters, standard
versus surface parameters (data not shown).
Further, a signi cant negative correlation with
age for skin elasticity [R5] and rmness [R7]
was observed by all three biomechanical evaluation devices. e viscoelastic component of skin,
described by parameter [R6] and represented by the
ratio of viscoelastic to elastic distension, i.e., Uv/
Ue, increased with age (see Figures 1 3).
Table 1 on Page 70 illustrates the correlation
coe cients between the three devices for the
biomechanical parameters measured as a function
of age. e best and highly signi cant correlation
was obtained on the skin rmness parameter, Ur/
Uf. e correlation between devices was lower for
the viscoelastic component, as illustrated here on
the coe cient of correlation for skin viscoelasticity, especially between the dermal torquemeter
and corneovacumeter. e higher variability of the
viscoelastic part of skin deformation may explain this
lower correlation. Signi cant correlation between the
three devices was observed for skin elasticity.
e
Derma Unit, Courage & Khazaka
Vol. 129, No. 4 | May 2014
Figure 1. Cutom eter results
Figure 2. Corneovacum eter results
Figure 3. Derm al torquem eter results
Vol. 129, No. 4 | May 2014
| 69
Tes t in g | C&T
Re s u lt s in Sk in C o lo r a n d
B r ig h tn e s s
Colorimetry measurements of luminosity and
ITA revealed a statistically signi cant evolution of
parameters with age, and the diminution observed
related to both UV exposed and non-exposed skin
on the cheeks and forearms (see Figure 4). e
evaluation of colorimetric parameters on macrophotographs con rmed skin color modi cation with
aging.
e diminution of luminosity and ITA , as well as
an increase of the yellow and red component of color,
Table 1. Correlation Coef
cients o f Three Biom echanical Properties Devices
Skin elasticity Ur/Ue [R5]

Cu
Cu
Dtm
Co
0.369,
p<
0.0001
0.416,
p<
0.0001
Dtm
Co
0.369,
p<
0.0001
0.416,
p<
0.0001
0.689,
p<
0.0001
0.689,
p<
0.0001
signi cantly changed. is increase of yellowish color

has been reported in the literature and linked to the
glycation and carbonylation of dermal proteins.10 In
relation, an increase in red color may be linked to
the unevenness of microcirculation and appearance
of the small vessels closer to the skin surface (see
Figure 5).
e skin complexion parameters measured by the
brillanometer were also signi cantly modi ed with
aging. A diminution of the peak height of specular
re ection and increase of the peak width of the peak
and concomitant decrease of the peak height (D/2)
in cross light, i.e., di use re ection, corresponds to
Skin viscoelasticity Uv/Ue [R6]

Cu
0.477,
p<
0.0001
0.531,
p<
0.0001
Dtm
Co
0.477,
p<
0.0001
0.531,
p<
0.0001
0.199,
p=
0.03
0.199,
p=
0.03
Skin rmness Ur/Uf [R7]

Cu
0.553,
p<
0.0001
0.525,
p<
0.0001
Dtm
Co
0.553,
p<
0.0001
0.525,
p<
0.0001
0.736,
p<
0.0001
0.736,
p<
0.0001
Cu = Cutometer; Dtm = Dermal Torquemeter; Co = Corneovacumeter
Figure 4. Evolutio n w ith age of param eters linked to skin color
lum inosity (L*) and ITA
Vol. 129, No. 4 | May 2014
Figure 5. Evolution w ith age of the param eters linked to skin

color lum inosity (L*), ITA , skin redness and yellow ness on the
cheeks, as evaluated from m acrop hoto graphs
lower, duller skin luminosity with

less radiance (see Figure 6). In
addition to the decrease of specular
re ection, a decrease in di use
re ection was observed, meaning
the skin was less radiant with age.
A strong positive correlation
was found between the di use
re ection and colorimetric parameters L* and ITA (see Table 2). e
correlation with specular re ection
was positive, but with a lower
correlation coe cient and limited
signi cance (p = 0.1). ese results
con rmed that the di use re ection had a strong impact on skin
luminance and radiance.
Flu o r e s c e n c e
Re s u lt s
Tryptophan uorescence is
a good marker for noninvasive
evaluation of the epidermal cell
proliferation rate. Since this
uorescence is associated with the
cell proliferation, its decline with
Vol. 129, No. 4 | May 2014
| 71
Tes t in g | C&T
Figure 6. Evolution w ith age of the param eters linked to the skin radianc e peak height
and w idth in specular re ection, and peak height in c ross light on the forearm s and cheeks
Table 2. Correlation Co ef
cients Betw een Skin Color (L*, ITA ) and Brightness
H, Height of the peak in

specular re ection
WH/2, Width of the peak in the

middle of height
D/2, Height of the peak in

cross light
Luminance, L*
0.162, p = 0.106
0.068, p = 0.503
0.697, p < 0.0001
ITA
0.165, p = 0.100
0.069, p = 0.495
0.653, p < 0.0001
age may indicate the reduced replicative capacity of

epidermal cells; i.e., replicative senescence.9 From this
study, the authors could conclude that photoaging
as well as intrinsic aging reduced cell renewal (see
Figure 7). In contrast, the uorescence of collagen
and elastin cross-links increased with age on the
forearms and face (temples), indicating an accumulation of AGEs (see Figure 8). e correlation between
di erent optical endpoints is illustrated in Table 3 on
Page 74.
A positive correlation was observed between the
skin complexion, evaluated by L* and ITA measurements, and epidermal turnover. Higher epidermal
turnover resulted in higher values for skin complexion. On the contrary, the accumulation of AGEs
with age correlated negatively with skin complexion
values. AGEs have been shown to be responsible for
the yellow discoloration of skin with aging. Indeed,
the negative correlation of AGEs and ITA , which
takes into consideration the L* and b* colorimetric
parameters, con rms this nding.10, 11
Epidermal proliferation, i.e., tryptophan uorescence, was also positively correlated with specular
and di use re ection, again indicating its importance in skin radiance. Finally, no correlation was
observed between AGEs and skin specular re ection;
on the contrary, a strong negative correlation was
found between di use refection and AGEs. Di use
re ection is diminished with aging due to a higher
absorption of light by chromophores present in the
skin.
A d d it io na l Pa r a m e t e r s
In regard to additional skin parameters, no
statistically signi cant variation of pH with age
was observed in any of the tested zones. Also, no
statistically signi cant evolution in hydration rate
was observed on the cheeks, although a signi cant
increase was observed on the forearms. Sebum excretion was strongly dependent on the age of volunteers,
with a signi cant decrease in the group of postmenopausal volunteers observed (see Figure 9).
Vol. 129, No. 4 | May 2014
Figure 7. Age-related dim inution of tryptophan

level on the forearm s and face (tem ples)
M e c h a nic a l v s . O p tic a l
Pr o p e r t ie s
To compare the mechanical and optical properties of skin, the authors used
the data obtained with the cutometer as
representative of skin's biomechanical
properties, since this device is generally
used. Table 4 on Page 75 shows the correlation between mechanical and optical
properties. e statistical comparison
demonstrates there are signi cant
correlations between the skin rmness
parameter, R7, and optical properties
linked to radiance, such as color and
brightness. A negative correlation was
found between the viscoelasticity of
skin, R6, and skin radiance, characterized by luminance and specular and
di use re ection. e correlation
between elasticity of the skin, R5, and
optical parameters was weaker and less
signi cant.
C o n c lu s io n
Figure 8. Evolution of norm alized uorescence

intensity w ith age for collagenase digestible collagen
crosslinks on the forearm s and fac e (tem ples)
Vol. 129, No. 4 | May 2014
e results of the present studies

con rm previously published data on
the evolution of skin elasticity and
rmness with age.12-17 Skin's optical
properties and their evolution throughout the aging process were evaluated,
and statistical correlation between the
di erent methodologies was performed.
A strong correlation was observed
between all devices used for the determination of biomechanical properties.
Optical properties including color and
brightness were strongly linked to age, as
was the formation of AGEs, as measured
by spectro uorimetry.
e parameters that correlated
positively with aging were the viscous
components of skin's biomechanical
properties, shown via an increase in
skin uorescence due to the glycation
of proteins, as well as an increase of
redness and yellowness. e negative
correlation with aging was observed for
skin elasticity, rmness, radiance and
turnover, and these parameters were
positively correlated with one another.
Skin rmness and elasticity was also
negatively correlated with the appearance of collagenase digestible cross-links
of collagen.
| 73
Tes t in g | C&T
Table 3. Correlation Co ef
cients Betw een Skin Color, Brightness and Fluorescence
Epidermal proliferation, tryptophan
Collagenase digestible crosslinks, AGEs
Luminance, L*
0.342, p < 0.0001
0.532, p < 0.0001
ITA
0.409, p < 0.0001
0.448, p < 0.0001
H = height of the
peak in specular
re ection
0.190, p = 0.029
0.137, p = 0.118
WH/2 = width of
the peak in the
middle height
0.231, p = 0.0077
0.148, p = 0.0914
D/2 = height of the

peak in cross light
0.294, p = 0.0006
0.657, p < 0.0001
Figure 9. Evolution of sebum excretion o n forehead as a function o f age
Interestingly, there was a signi cant negative

correlation between skin radiance, L* and ITA , and
the formation of AGEs, with a simultaneous positive
correlation of AGE and yellow color. Negative correlations also were found for epidermal skin turnover
and increased red and yellow colors.
Finally, a decrease in skin luminance and radiance
was positively correlated with the decrease in epidermal turnover. us, this overview of the evolution of
di erent parameters with aging demonstrates how
all these parameters are tightly linked together and
modi ed during the aging process.
Acknowledgements: e authors wish to acknowledge

Catherine Bonnaud-Rosaye and Nadine Duc-Sikora for their skilled
technical contributions.
References
1. BC Murray and RR Wickett, Correlation between Dermal Torque
Meter, Cutometer and Dermal Phase Meter measurements of
human skin, Skin Res Tech 3, 2 101 106 (1997)
2. N Krueger, S Luebberding, M Oltmer, M Streker and M Kerscher,
Age-related changes in skin mechanical properties: A quantitative evaluation of 120 female subjects, Skin ResTechnol 17, 2
141 148 (2011)
3. GE Pi rard, EEMCO Guidance to the assessment of skin color,
J Eur Acad Dermatol Venereol 10, 1, 1 11 (1998)
Vol. 129, No. 4 | May 2014
Table 4. Correlation of M echanic al and Optic al Skin Properties

Luminance, L*
ITA
H, height of the
peak in specular
re ection
WH/2, Width of
the peak in the
middle of height
D/2, Height of
the peak in the
cross light
Cutometer,
Ur/Ue [R5]
0.240
p = 0.010
0.170
p = 0.072
0.153
p = 0.109
0.169
p = 0.077
0.169
p = 0.077
Cutometer,
Uv/Ue [R6]
0.312
p = 0.007
0.264
p = 0.004
0.262
p = 0.005
0.218
p = 0.021
0.258
p = 0.006
Cutometer,
Ur/Uf [R7]
0.304
p = 0.001
0.227
p = 0.015
0.206
p = 0.031
0.204
p = 0.032
0.226
p = 0.017
4. JC Pittet, C Musnier, SC Jacobs, M Lebel, JM Baret and

P Beau, Evaluation of the property of the skin to absorb and
re ect the light, New instrumental method and visual sensory
evaluation, Proceedings of the 22nd IFSCC Congress, Edinburgh 485 490 (2002)
5. A Petitjean, JM Sainthillier, S MacMary, P Muret, B Closs,
T Gharbi and P Humbert, Skin radiance: How to quantify?
Validation of an optical method, Skin Res Technol 13, 2 8 (2007)
6. M Sabadotto, O Freis, G Perie and A Rathjens, The Corneovacumeter: A new device for evaluation of biomechanical
properties of the skin, J Cosmet Sci 5, 354 355 (2011)
7. D Qu et al, Correlation age and quantifying product ef cacy on
human skin using novel viscoelastic parameters, Proceedings
IFSCC 1 6 (2007)
8. V Gillon, G Perie, S Schnebert and G Pauly, A new method
for contactless in vivo quantitative measurement of stratum
corneum gloss attributes: In uence of natural active ingredients,
in The Essential Stratum Corneum, R Marks, JC L v que and
R Voegeli, eds, Taylor & Francis, Philadelphia (2002) 331 334
9. GN Stamatas, RB Estanislao, M Suero, ZS Rivera, A Khaiat
and N Kollias, Facial skin uorescence as a marker of the skin's
response to chronic environmental insults and its dependence
on age, Br J Dermatol 154, 125 132 (2006)
10. H Ohshima et al, Melanin and facial skin uorescence as markers of yellowish discoloration with aging, Skin Res Technol 15,
496 502 (2009)
Vol. 129, No. 4 | May 2014
11. Y Ogura et al, Dermal carbonyl modi cation is related to the

yellowish color change of photo-aged Japanese facial skin,
J Dermatol Sci 64, 45 52 (2011)
12. T Ezure and S Amano, In uence of subcutaneous adipose tissue
mass on dermal elasticity and sagging severity in lower cheek,
Skin Res Technol 16, 332 338 (2010)
13. T Ezure and S Amano, Involvement of upper cheek sagging in
nasolabial fold formation, Skin Res Technol 18 259 264 (2012)
14. N Krueger, S Luebberding, M Oltmer, M Streker and M Kerscher,
Age-related changes in skin mechanical properties: A quantitative evaluation of 120 female subjects, Skin Res Technol 17
141 148 (2011)
15. H Ohshima et al, Use of Cutometer area parameters in evaluating age-related changes in the skin elasticity of the cheek, Skin
Res Technol 19 e238-242 (2013)
16. A Firooz et al, Variation of biophysical parameters of the skin
with age, gender and body region, Scienti c World J 1 5 (2012)
17. PA Wendling and G Dell'Acqua, Skin biophysical properties of
a population living in Valais, Switzerland, Skin Res Technol 9
331 338 (2003)
18. C Arce-Lopera, T Igarashi, K Nakao and K Okajioma, Effects of
diffuse and specular re ections on the perceived age of facial
skin, Optical Rev 19 167 173 (2012)
| 75
Fo r mul at in g | C&T
Approaches and Issues in
Powder Formulations
Peter Tsolis
The Est e Lauder Companies, Melville, NY
Gil Sahagun
Mana Products, NYC USA
KEY WORDS
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hough many people associate color cosmetics with anhydrous lipsticks

and emulsion foundations, powder formulations have always been a
core product category that drives brand growth and consumer loyalty.
e U.S. powder market alone is set to grow by almost 7% by the end of
scal 2014.1 is category has been steadily gaining share in the makeup
market, with new technologies entering the market in both packaging and
raw materials. Currently, powder formulations contribute approximately
12% to annual global makeup sales.
e skill set for complete powder formulating can only come from
years of experimentation and development using a variety of powder-based
products. It requires thorough knowledge of raw materials, color matching techniques and processing. Besides being a formulator, this is one case
where the formulator must also uno cially play the role of a packaging
analyst to ensure compatibility with the proposed components. Whether
the powder-based product is loose or pressed, its stability in a packaging
component can dictate its performance.
As noted, powder cosmetic products (both pressed and loose powders)
include foundations, eyeshadows, primers, blushes, bronzers and all-over
shimmer powders. ough the raw materials used in them o en are similar,
the processing, texture, packaging, applicators and application, along with
color development, vary for each type; these are described here.
Ra w M a t e r ia ls a n d Us e s
Powder formulations involve the blending of pigments, llers, and dry
and wet binders to develop a uniform product. e coverage and shade
provided by a product is de ned by the levels of organic dyes, iron oxides
and titanium dioxide in the formula as well as processing techniques.
Dry binders: Dry binders are a necessity to powder formulations. ey
compact easily and use their adhesive properties to assist other ingredients
in compacting. Some common dry binders are polyethylene, kaolin and
fatty acid derivatives including their metallic soaps, such as zinc stearate.
Surface treatments: Surface treatments on llers like talcs, sericites and
micas are used to provide smooth application while assisting with wear
and adhesion to the skin. eir use will change the overall coverage of the
makeup. e addition of surface-treated materials may reduce the need

Vol. 129, No. 4 | May 2014
Mibelle AG Biochemistry, 5033 Buchs/Switzerland, Phone +41 62 836 17 31
Snow Algae Powder

Key to skin's longevity
Snow Algae Powder stimulates the longevity gene Klotho and activates AMPK,
which is a master sensor of cell energy. Through the activation of Klotho and
AMPK a calorie restriction-mimetic anti-aging effect can be achieved in the
skin for the very first time.
Snow Algae Powder is based on the extract of a unique algae that is able to
grow on glaciers and permanent snow. In these extreme environmental conditions the algae produces valuable stress response molecules that protect the
youthfulness of the skin.
Distributed in the USA by:

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for wet binders in the system, as they can be readily
incorporated into the formula. Some popular surface
treatments are based on alkyl silane and dimethicone
chemistries. ese treatments impart greater hydrophobicity and skin adhesion to the formula, as well as
a smoother and more spreadable application. Other
treatments may include phospholipids or lecithin,
which provide similar bene ts and deliver a more
lubricious end feel.
Po w d er- b ased
fo rm ulatio ns c o m e
w ith their o w n set o f
c o nc erns that are
d ifferent fro m typ ic al
skin c are o r m akeup
fo rm ulatio ns.
Wet binders: Wet binders are added to wet the
pigments in a formula and help dictate the overall
feel of the product. ey are essential in ensuring
pigment dispersion, and commonly are the ingredient/phase to adjust when formulation issues such as
uneven application arise. ese materials are usually
silicones, esters or synthetic hydrocarbons. ey may
also include solid materials that are not always liquid
at room temperature, such as waxy polymers and
esters.
Fillers: Fillers are used to maintain product texture and compressibility throughout a shade family,
as well as to provide improved wear bene ts, depending on the surface treatment. Talc, mica and sericite
are predominantly used.2 Ingredients that prevent
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caking, such as silica, may also be added to loose

powder formulas, in cases where texture di erences
are desired.
St a b ilit y C o n c e r n s
Powder-based formulations come with their own
set of concerns that are di erent from typical skin
care or makeup formulations. Many problems relate
to pressing that can, therefore, only be seen once the
powder is pressed; for example, dusting a er touching the surface or glazing, i.e., a shiny appearance
a er the applicator is used. Others include stability
concerns that develop over time, such as the clumping of particulates or di culty with pick-up, making
the product too hard to press. Visually inspecting the
surface and removing some product to apply it will
reveal the product's consistency and aesthetics. When
a glaze develops or product becomes overly rigid, it
is o en due to binder migration through the tablet
toward the surface of the product.
In the lab, potential stability problems can be
identi ed by traditional tests, whereby the product is
exposed to a variety of extreme temperature conditions. Further tests include drop tests and shaker
stations to challenge the tablet's ability to survive
shipping conditions. While binding the product
correctly will result in good stability, the formulator
must balance this with production concerns related
to di culty in compressing the tablet. Using both a
dry binder such as zinc stearate and an anti-caking
ingredient like silica is usually helpful for balancing
the powder and avoiding unexpected pressing issues
during production. Note that the described tests may
also be package-speci c; i.e., what works in a oneinch round pan may not work in a four-inch square
pan scenarios that may be proposed for a special
marketing program.
Pr o c e s s B a s ic s a n d
Eq uip m e nt
Many powder-based formulation issues can be
avoided by using the same process and systems in
production that were successful in the laboratory.
Processing powder products typically involves the
steps of de-agglomerating particulates via pulverizing/milling, adding liquids and other powders, and
blending.3 e formulator must e ciently advance
through this process in as few steps as possible.
Processing equipment in the lab is o en limited
to blenders, co ee and spice grinders, and simple
blenders. A blender or co ee grinder, which contains
spinning blades, is supposed to simulate pulverizers
and mills in production. ese appliances are similar
to hammer mills, which use a hammer to break down
and deagglomerate pigments. While these appliances
Vol. 129, No. 4 | May 2014
process pigments, the production mill does a more

uniform and consistent job, leading to undispersed
pigments in laboratory batches. e powder shades
in the lab would therefore neither be fully developed
nor attain maximum color distribution throughout
the product. us, with all manufacturing equipment,
color development can be a ected if the mixing is
not the same. In relation, temperature also must be
controlled, especially in smaller equipment where
excessive heat can be generated, extending product
processing and causing agglomeration.
Formulators who use multiple types of blenders
during development can, therefore, expect di erent
results in the pressing and color development of
powders. Correlating what a formulator does in the
lab to what the results are in production can take
some e ort, but a er moving a product from the
lab to production, standardizing the techniques will
minimize inconsistencies from one formulator to the
next and generate successful production processing
results at least for that formula family.
Also during scale-up, it is important to consider
the potential for loss of ingredients; e.g., of liquid
binder, though the transfer and spraying apparatus.
Accounting for loss will help to maintain the proper
powder to liquid ratio. Adding extra liquid binder
Vol. 129, No. 4 | May 2014
will keep the formula balanced so it compresses in a

consistent manner. Another consideration is properly
loading the powder in the mixing vessel to ensure
the binder is sprayed on the powder and not on the
walls and impeller.4 Binder that is lost in the lines or
stuck on the walls can make the powder drier and
looser, and will a ect the pressing and stability of the
product.
C o lo r C o n s is t e n c y a n d
Pe a r l Le v e l
Achieving the nal powder color and coverage
is key, so to fully develop shades and minimize
undispersed pigments, processed extenders can be
used; for example, Red 7 in talc extender. Qualitycontrolled pre-pulverized or jet-milled pigment
extenders can help the formulator develop shades in
the lab that are consistent to the pilot and production batches. While this approach may not always
be the easiest or most cost-e ective, it is the most
controlled. Such extenders are usually not the full
formula; they typically contain the pigment at a
relatively high level, mixed and milled with llers
used in the product family and others. ey do not
easily separate, so storage and long-term stability are
rarely issues.
| 79
During shade-matching, it is also important to
maintain consistent levels or percentages of pearl
and mica from batch to batch. is will give the
consumer a more consistent feel and application, as
it helps to dictate the spreadability of the product.
Formulators should also maintain similar-sized
pearl particles from shade to shade within a family
to maintain texture and application parameters. As
with most particulates, smaller-particle pearls feel
smoother and lay more evenly, compared with larger
pearl particles that tend to be raspy and lay unevenly.
When employing borosilicates and synthetic uorophlogopites, it is advisable to use them sparingly to
maintain the feel as much as possible. Milling these
special materials will reduce grit as well as their ability to provide the special light e ects for which they
were added; further, milling these types of pearls is
not cost e ective.
Maintaining a consistent product while colormatching is usually a big hurdle, and formulators
should know the threshold of their formula when
it comes to color adjustments. If the adjustment
addition is equal to 20% of the batch, for example,
the overall adjustment also should include corrective measures for liquid and dry binders, as well as
preservative, to bring the formula back to 100%.
Is s u e s in Pa c k a g in g
Most technical issues with powders involve
processing and packaging, since problems that arise
during formulating and pressing are easily corrected
at a lab scale. erefore, packaging components
should be consistent throughout the development
phase and into manufacturing, speci cally in relation
to the shape and depth of pan.
e formula is built around packing and binding
particulates and solvents in a stable formula that is
speci c to a component size. If a formulator uses a
relatively large pan when lling and pressing in the
lab and suddenly changes to a smaller pan, glazing
in the powder is sometimes the result. If the opposite
occurs and a formulator switches to a larger-sized
pan, the powder may not pass the drop test, forcing
the formulator to press the powder harder, in turn
compromising its application and feel. ere is also
a concern when it comes to the shape of the pan. A
formula can sometimes act di erently if the formula
was created in a round pan in the lab and the nal
pan is square, for example; in this case, production
will usually see the pressed pieces breaking at the
corners and edges.
When it comes to loose powders, an obvious issue
is when dispensing ori ces in the si er are not com-
Vol. 129, No. 4 | May 2014
patible with the formulation. e size and number of
ori ces play a role in proper dispensing. Changing
the si er will a ect the delivery of the product onto
the applicator and onto the skin. In addition, the
applicator is a key component to fully deliver the
bene ts of the product. Whether the applicator is a
brush or sponge, its early identi cation for formulation compatibility will avoid problems during launch.
Applicators can help to develop the product's proper
shade and deposition on the skin, as well as minimize
glazing or dusting. And although brush sizes may be
the same, whether the bristles are natural or synthetic
can impact optimal application. Neither is necessarily
superior; each has bene ts depending on the formula
and package.
C o n c lu s io n
Makeup companies will continue to focus e orts
on new innovations and formulations in the powder
category. To attract new consumers, new packaging
and processing techniques will bring novel ways
to press design shapes or personalized logos on
products. Furthering formulations with additional
consumer bene ts while maintaining proper stability
will continue to make this a growing market.
References
1. US Make-up Retail Sales Fiscal 2014, NPD Beauty Trends report
(2014)
2. J Hollenberg, Color Cosmetics, ch 26 in Harry's Cosmetology,
8th Edition, Chemical Publishing Co., Revere, MA USA (2000)
pp 560 588
3. H Rumpf, Science of Agglomeration, Chem Tech 45 (1) 1 11
(1974)
4. D Buell, K Barclay, P Block, J Junker, B Victor, D Melenkevitz
and D Yacko, The Manufacture of Cosmetics, ch 35 in Harry's
Cosmetology, 8th Edition, Chemical Publishing Co., Revere, MA
USA (2000) pp 787 874
GUES T CON TRIB UTOR

Gil Sahagun is currently manager
for the powder department of
Mana Products in New York.
He has worked at several major
nished goods companies and
developed countless powderbased formulations for the mass
and prestige cosmetic markets.
Vol. 129, No. 4 | May 2014
!
"
!
!
)
" ##$ %&

$# $* %+, $*
' (
Novel Azobenzene Com pound to
Extend and Reactivate

UV Protection
presentation
Jin-Ye Wang, PhD, and Shengyong Geng

Shanghai Jiao Tong University, Shanghai, China
Editor's note: While Cosmetics & Toiletries acknowledges that animal testing is a sensitive
issue to many readers worldwide, it is important to note that in China, where these authors
conducted their research, it is currently legally required. All animal experiments involved in
this study were performed in accordance with Regulations for the Administration of A airs
Concerning Experimental Animals of China.
KEY WORDS
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V radiation can be absorbed by di erent chromophores in the skin,

and absorption of UV photons by these chromophores results in different photochemical reactions and secondary interactions involving
reactive oxygen species (ROS), which result in harmful e ects.1 UV radiation is comprised of 98% UVA (320 400 nm) and 2% UVB (290 320 nm).2
UVB radiation is mainly responsible for the most severe damage: acute
damage such as sunburn and long-term damage, including cancer. It has a
direct impact on cell DNA and proteins. Unlike UVB, UVA radiation is not
directly absorbed by biological targets, but can still dramatically impair cell
and tissue functions.3, 4
An ideal sunscreen should provide uniform UVB/UVA protection, and
remain unchanged a er UV irradiation.5 Another consideration is safety;
this is a fundamental point in the eld of cosmetics. Skin presents a signi cant barrier for most substances applied. Typically, one wants UV lters
to stay on the surface or to not permeate lower than the upper epidermis;
certainly not to the vascular dermis.
In the present study, two photosensitive compounds, i.e., 4-cholesterocarbonyl-4'-(N,N'-diethylaminobutyloxy) azobenzene (ACB) and
4-cholesterocarbonyl-4'-(N,N,N-triethylamine butyloxyl bromide) azobenzene (CAB), were synthesized. e phototoxicity of the two compounds
was rst evaluated in animal models. e cytotoxicity a er combined either
ACB or CAB into liposomes was then compared with NIH3T3 cells. In
addition, their skin permeation ability was assessed. Finally, ACB, the safer
of the two, was studied in greater depth for its stability under UV and visible light irradiation, as well as its UVA and UVB protective e cacy.
Ex p e r im e n t a l D e s ig n
Materials: Egg phosphatidylcholine (PC) and cholesterol (CHOL)
were purchaseda. e commercial sunscreen b 4-tert-butyl-4'-mea, e
b
PC, CHOL, and 8-methoxypsoralen, Sigma (USA)

Avobenzone, King Jack Int.

Vol. 129, No. 4 | May 2014
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w w w.lucasm eyerco sm et ics.co m
thoxydibenzoylmethane also known as avobenzone
(C20H 22O3) also was obtained. e cream substrate
used was provided gratisc, and water-soluble CdTe630
quantum dots (QDs) were purchasedd. Finally, ACB
(C48H 71N3O3, Mw 737) and CAB (C50H 76N3O3+, Mw
766) were synthesized.6 8
Phototoxicity: A phototoxicity assay was performed with guinea pigs, according to Hygienic
Standard for Cosmetics.9 e sample was prepared as
1 mg/mL in distilled water for CAB, and 1 mg/mL in
re ned oil for ACB; 0.05% of 8-methoxypsoralen e was
used as the positive control.
ACB lip o so m es
und erw ent re versib le
trans- c is iso m e rizatio n.
Liposome preparation: Two types of liposomes
were evaluated for di erent purposes: one for
UV protection and another for skin permeation,
which was loaded with QDs. e former included
traditional liposome (PC-liposome), avobenzonecombined liposome (avobenzone-liposome),
ACB-combined liposome (ACB-liposome) and
CAB-combined liposome (CAB-liposome). e latter
included QDs-loaded PC liposome (PC-QDs); QDsloaded ACB liposome (ACB-QDs); and QDs-loaded
CAB liposome (CAB-QDs). e preparation of liposomes was carried out using a standard sonication
method under nitrogen, as described previously.10, 11
Cytotoxicity of blank liposomes: NIH3T3 cells
were cultured in DMEM supplemented with 10%
fetal calf serum. en cells were seeded on the
c
Cream substrate, Shanghai Huashan Cosmetics Co., Ltd.

QDs, Shen Zhen ZhongDS Investment Co., Ltd.
f
Microplate reader, Stat Fax 2100
d
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96-well plates at a density of 5 104/mL with a

humidi ed condition at 37 C and 5% CO2 for 24 hr.
e culture medium was then replaced with medium
containing serial dilutions of PC, ACB and CAB in
various liposomes. e concentration of cholesterol
in PC-liposome group was maintained the same
as other groups. A er the 24 hr incubation, the
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was conducted by a
microplate reader f at 490 nm. e higher absorbance
indicates a higher cell vitality.
Photo-isomerization of ACB in liposomes: ACBliposome was irradiated by a 400 W high-pressure
Hg lampg attached with a bandpass lter (l T =
275 400 nm) to obtain complete isomerization. e
recovery of ACB was carried out using a uorescent
lamp. e spectrum was measured using a UV Vis
spectrophotometerh.
UVA/UVB protection of ACB-liposome in cream
substrate: e ACB-liposome and cream substrate
was applied at an amount of 20 mL cm -2 (~ 0.06 mg
ACB) and 2 mg cm -2, respectively, onto tapesj (1 cm
4 cm), which were then dried for 30 min at 37 C. e
UVB protection e ect was evaluated according to the
QB/T 2410-1998 standard method.12 Brie y, the average absorbances at 280 nm, 290 nm, 300 nm, 310 nm
and 320 nm were measured using ultraviolet UV
spectrophotometer and calculated as Ai. e average
absorbance Ai was repeated ve times and recorded
as . When
0.5, the cream could be considered to
provide UVB protection. e UVA protective e ect
was assessed by the critical wavelength method.13
In vivo UV protection: In vivo, the UV protection
e cacy was evaluated both with SD rats (150-200 g)
and BALB/c nu/nu mice ( ve to six weeks). Four
test sites, 1-cm 2 per site, were selected on the back
of shaved rats or nude mice (Figure 1). e four
samples including avobenzone-liposome, ACBliposome, PC-liposome and avobenzone solution,
containing 5% (v/v) glycerol were spread uniformly
on the four testing sites, respectively. e samples
were allowed to dry for 15 min before UV irradiation g. e skin was irradiated for 5 hr or 3.5 hr at a
distance of 15 cm from the light source. In order to
ensure that UV absorption was the same, the applied
volumes of avobenzone-liposome and ACB-liposome
were determined according to their UV absorption at
360 nm.
In vivo permeation: For permeation studies, the
treatment of BALB/c nu/nu mice was the same as
described for UV testing. Four areas on the back
were selected and naked QDs, PC-QDs, ACB-QDs
g
h
j
Hg lamp, LCE-9, Zhengzhou, China

Spectrophotometer, U3010, Hitachi
Transpore tape, 3M
Vol. 129, No. 4 | May 2014
Figure 1. Experim ental design
and CAB-QDs in 5% glycerol (v/v) were applied at

a QD concentration of 20 nmol cm -2. Mice (n = 3
per group) were sacri ced at 0, 2, 4, 8, 12 and 24 hr
by injection, and the dorsal skins were rinsed three
times with wet gauze. Skin samples were embedded
in TEK OCTk, frozen at 20 C, and sectioned at 5 m
by cryostatm . e sections were examined using a
uorescence microscopen .
Re s u lt s
Phototoxicity and cytotoxicity: Dermal irritation
studies revealed that both ACB and CAB, in their
k
m
n
TEK OCT, Sakura FineTek Inc.

Cryostat CM3050S, Leica
Fluorescence microscope BX51, Olympus
Vol. 129, No. 4 | May 2014
pure compound state, did not show phototoxic properties when applied topically (data not shown). e
cytotoxicity of blank PC-liposome, ACB-liposome
and CAB-liposome to NIH3T3 cell lines was compared, and for neutral ACB-liposome, no signi cant
cytotoxicity was observed. is was the case even
when the concentration of ACB was increased to
1.2 mol/mL; i.e., the cytotoxicity of ACB-liposome
was similar to the neutral PC-liposome. Although
positive CAB-liposome showed some toxicity
compared with ACB-liposome, it is safer than the
commercial positive liposome N-(1-(2, 3-dioleoyloxy) propyl-N,N,N-trimethylammonium mesylatep.
p
DOTAP transfection reagent, Roche Diagnostics
| 87
It is clear that ACB was better than CAB from a safety
point of view. erefore, the authors then investigated
the possible application of ACB as a sunscreen.
ACB-liposome isomerization and stability
under irradiation: As noted, photostability is a key
parameter for e ectiveness in commercial sunscreen products, and as shown in Figure 2, ACB in
liposomes underwent reversible trans-cis isomerization and this recovery process could be repeated
at least 10 times. Unlike the commonly used UVA
lter avobenzone,14 the azobenzene structure in
ACB is stable and does not decompose under light
irradiation. e transmission electron micrographs of
ACB-liposome before and a er 10 cycles of periodic

UV-Vis irradiation showed that the structure of
ACB-liposome also maintained its original form (see
Figure 3). erefore, periodic UV and visible light
irradiation could not a ect the photo-isomerization
or structure of ACB-liposome.
ACB-liposome UVA/UVB protection with cream
substrate: e critical wavelength method, rst
established by Di ey in 1994, is based on instrumental measurements to evaluate UVA protection.13 e
critical wavelength (lc) is the wavelength at which
the area under the spectrum from 290 nm to lc is
90% of the integral of the absorbance spectrum from
290 nm to 400 nm. As
shown in Figure 4a, the
total area of the spectrum
Figure 2. Isom erization of ACB-liposom e under UV-Vis
from 290 nm to 400 nm is
irradiation; isom erization rate (%) = 100 At/ A 0 , w here A 0 is the
75.299 nm * Abs; thus, 90%
absorbanc e at the m axim um absorption w avelength (lm ax) of
of this area is 67.769 nm *
ACB-liposom e in the initial state, and At is the absorbance at lm ax at
Abs. According to Figures
different irradiatio n tim e po ints.
4b and 4c, the area of the
spectrum from 290 nm to
390 nm and from 290 nm
to 389 nm is 68.434 nm *
Abs and 67.706 nm * Abs,
respectively.
So the lc of the ACBliposome, when mixed with
the cream substrate, was
between 389 nm and 390
nm, i.e., lc > 370 nm. is
satis es the UVA measurement regulations of the U.S.
Food and Drug Administration that allow only
products with lc > 370 nm
to be labeled as broad
spectrum. 15 For UVB, the
average absorbance of
the ACB-liposome, when
mixed with the cream
substrate, was between 0.5
Figure 3. Transm ission electron m icrographs of ACB-liposom e
and 1.0, indicating this
a) before and b) after ten c ycles of p eriodic UV and visible
formulation provided UVB
irradiation; voltage w as set at 2 00 kV and scale bar = 100 nm .
protection according to the
QB/T 2410-1998.
UV protection in vivo:
Erythema formation
measurements were used
to evaluate the UV protection levels of sunscreens
in vivo.16 e e cacy of
ACB-liposome in animal
models is shown in Figures
5 and 6 on Page 90, respectively. As seen in Figure 5a,
Vol. 129, No. 4 | May 2014
a er UV irradiation for 5 hr, the surrounding area for the PC-liposome showed
much more obvious erythema than did the ACB-liposome, avobenzone-liposome
or avobenzone. Figure 5b showed that the same erythema was not so obvious
one day a er irradiation. In Figure 5c, two days a er UV irradiation, erythema
continued to become even more inconspicuous, although festering became
apparent. ree days a er UV illumination, festering was not observed with the
ACB-liposome or avobenzone-liposome, while skin coated with PC-liposome
and avobenzone showed serious festering (see Figure 5d); thus, both ACB and
avobenzone showed UV protection.
In Figure 6a, a er UV irradiation for 3.5 hr, the total skin area became red
Continued on Page 92
Figure 4. UVA p rotec tive effect of ACB-lip oso m e w hen m ixed

w ith the cream substrate, as evaluated by critic al w avelength
m ethod; area of the spectrum a) from 290 40 0 nm , b) from 290
390 nm , and c) from 290 389 nm .
Vol. 129, No. 4 | May 2014
| 89
Figure 5. Erythem a and am bustion after UV irradiation for 5 hr, then observed after: a) 0
days, b) 1 day, c ) 2 days and d) 3 days; a = avo benzone-liposom e; b = ACB-liposom e; c = PCliposom e; d = avobenzone; reproduced w ith perm issio n from The Royal Society of Chem istry.
Figure 6. The skin states observed at: a) 0 days, b) 4 days, c ) 5 days and d) 6 days after UV
irradiation for 3.5 hr; a = avobenzone-lipo som e; b = ACB-liposom e; c = PC-liposom e; and
d = avobenzone.
Vol. 129, No. 4 | May 2014
Figure 7. Fluoresc ent im ages of skin

sections after applied sam ples onto the dorsal
skin surface of nude m ic e fo r 2 hr; a) naked
QDs; b) PC-QDs liposom e; c) ACB-QDs
liposom e; and d) CAB-QDs liposom e; sc ale
bars = 100 m ; reproduc ed w ith perm ission
fro m The Royal Soc iety of Chem istry.
but the area treated with PC-liposome and avobenzone

seemed more obvious. Four days a er UV irradiation,
erythema was not obvious, as shown in Figure 6b, but
slight skin ambustion could be observed in the areas
without any coating or those coated with PC-liposome (see
Figure 6c). Six days a er UV irradiation, no skin ambustion was observed in the areas coated with ACB-liposome,
avobenzone-liposome and avobenzone, while the skin
coated with PC-liposome showed festering as serious as
the area without any coating (see Figure 6d). Moreover,
skin peeling could be observed in the areas coated with
avobenzone-liposome and avobenzone. erefore, both
ACB and avobenzone displayed UV protection, but ACB
proved to be much more e ective.
In vivo permeation: As described, the permeation ability of the three kinds of liposomes into nude mouse dorsal
skin was evaluated by loading QDs as uorescent probes.
As seen in Figures 7 and 8, on Page 94, naked QDs and
QDs-loaded neutral liposomes, i.e., PC-QDs and ACBQDs, broke through the barrier of the stratum corneum
but stayed in the upper epidermis; they did not reach
the dermis even a er 12 hr. On the contrary, the cationic
liposome CAB-QDs not only penetrated the barrier of the
stratum corneum, they also gradually reached the deeper
epidermis, close to dermis, a er 8 hr (see Figure 8d), and
near the vascular dermis a er 12 hr. e authors propose
that the cationic liposome could bind to negative stratum
Vol. 129, No. 4 | May 2014
Figure 8. Fluoresc ent im ages of skin sections after applied

sam ples onto the dorsal skin for 8 hr; a) naked QDs; b) PC-QDs
liposom e; c) ACB-QDs liposo m e; and d) CAB-QDs liposom e;
sc ale bars = 100 m ; reproduced w ith perm ission from The
Ro yal Society of Chem istry.
corneum, leading to an enhanced penetration into and through the skin.11, 17

Since the neutral ACB-liposome stayed on the skin surface and did not penetrate
into the vascular dermis, it was considered appropriate for sunscreens.
C o n c lu s io n s
As noted, the photostability of sunscreens is a key parameter since a sunscreen
that loses its capacity to block UV radiation provides a clear risk of damage to the
user. And as mentioned, the ACB-liposome not only protected skin from damage
caused by UV irradiation but also had good stability both morphologically and
chemically; i.e., UV irradiation did not decompose ACB. One might imagine that
when ACB-liposome treated skin is exposed to UV light outdoors, the chemistry
changes from trans to cis by absorbing UV light. en as soon as they move
indoors, it returns to the trans form and recovers its UV-absorbing ability again
when the user steps out.
According to the present study, this process can be repeated at least 10 times,
suggesting the potential for users to save time and cost. In addition, as an active
sunscreen ingredient, the stability and photo-isomerization of ACB-liposome was
evaluated when mixed with other cosmetic additives such as propylene glycol and
glycerol. Results showed that in the presence of a mixture of 20% propylene glycol
and 5% glycerol, the repeated photo-isomerization upon UV irradiation was
the same as it was for the liposomal emulsion without additives.18 What's more,
when cosmetic actives such as ascorbic acid and catalase when incorporated
into the ACB-liposome, their release could be controlled by UV irradiation.10 As
a conclusion, this technology is promising for the development of reusable and
multifunctional sunscreen formulations.
Vol. 129, No. 4 | May 2014
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Vol. 129, No. 4 | May 2014
| 95
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75
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A&E Connock Ltd.

sales@connock.co.uk
www.connock.com
info@activeorganics.com
www.activeorganics.com
51
15
61
C4
83
80
29
57
67
92
65
37
87
Air Products & Chemicals, Inc.

www.airproducts.com/skin
81
Alluredbooks
books@allured.com
www.Alluredbooks.com
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1
info@aristaindustries.com
www.aristaindustries.com
31
35
Beraca Ingredients
www.beraca.com
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Berj , Inc.
berje@berjeinc.com
www.berjeinc.com
71
Bioland Ltd.
bioland@biolandkorea.com
www.biolandkorea.com
93
Biosil Technologies, Inc.

www.biosiltech.com
Centerchem, Inc.
64
cosmetics@centerchem.com
www.centerchem.com
Chemsil Silicones, Inc.
19
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CIDP
www.cidp-cro.com
79
Clariant International Ltd.

info@clariant.com
www.personalcare.clariant.com
Coast Southwest, Inc.

info@coastsouthwest.com
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Consumer Product Testing Co.
39
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Corum, Inc.
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www.corum.com.tw
Cosmetics & Toiletries Summit
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85
FD
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Extracts & Ingredients
Grant Industries
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Hallstar
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Honeywell International
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Ichimaru Pharcos Co. Ltd.

gifu@ichimaru.co.jp
www.ichimaru.co.jp
Idea Tests
r.carer@groupeideatests.com
www.groupeideatests.com
IFSCC/Paris 2014
ifscc@clq-group.com
www.ifscc2014.com
Ikeda Corp.
info@ikeda-america.com
www.ikeda-corp.co.jp
Innospec Ltd.
america-pc@innospecinc.com
www.innospecinc.com
Innovadex
www.ulprospector.com
INOLEX, Incorporated
cheminfo@inolex.com
www.inolex.com
49
45
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59
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Lonza, Inc.
Naturex
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Nikko Chemicals Co. Ltd.

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NY SCC/Antioxidant Symposium
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Pilot Chemical Co.

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Presperse Corporation
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sch lke, Inc.

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Sederma France
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Silab
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Sinerga
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Solabia Group
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Summit Events/Anti-aging Conf.,

London
The Center for Professional

Innovation & Education
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info@lipoid-kosmetik.com
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Lipotec, LLC
info@mibellebiochemistry.com
info@summit-events.com
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Mibelle AG Biochemistry
(p. 14)
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dfondots@morretec.com
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info@lucasmeyercosmetics.com
www.lucasmeyercosmetics.com
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(p. 50)
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Lubrizol Advanced Materials, Inc.

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