Content >

11-2010

English Edition
International Journal for Applied Science
• Personal Care • Detergents • Specialties

Penetration of Topically
Applied Substances

UV-Induced Skin Erythema

Development of Skin Models

Tetrapeptide for Elasticity

and Tightness Enhancement

Grooming Practices of Hair

Fragrance Ingredients

Stem Cells

EU-Ecolabel
for Detergents
 Content >

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COSMETICS
J. Lademann, H.-J. Weigmann, S. Schanzer, M. Meinke,
W. Sterry, A. Patzelt:
Analysis of the Penetration of Topically Applied Substances
into the Human Skin . . . . . . . . . . . . . . . . . . . . . . . . . . 2
G. Dell’Acqua, K. Schweikert, G. Calloni:
Stimulating and Protecting Skin Immunity to
Decrease UV-Induced Skin Erythema . . . . . . . . . . . . . . . 10
J.W. Fluhr, R. Darlenski:
Development of Skin Models –
Sensitive Skin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
M. Mateu, E. Cañadas, J. Cebrián, C. Carreño, N. Almiñana,
R. Delgado:
A New Tetrapeptide for Elasticity and Tightness
Enhancement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Y. Kamath, V. Gode, N. Bhalla, B. Kalghatgi, S. Mhaskar:
Effect of Grooming Practices on Physical Characteristics
of Hair . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
S. Subramanian, A, Wingenfeld:
Fragrance Ingredients with Growth Inhibiting Activity . . . 40
L. Cattuzzato, C. Sanchez, A. De Pooter, M. Puginier,
S. Dumont:
Complementary In Vitro Models to Investigate the Mode
of Action of Active Ingredients on the Protection of
Extracted Epidermal Stem Cells Against Different Types
of Stresses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
DETERGENTS
M. Gast:
Revision of the EU-Ecolabel
for Detergents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
SPECIALTIES
P. Morganti, Yuan-Hong Li:
Wellness and Beauty Outside in:
China West – Working Together . . . . . . . . . . . . . . . . . . . 59
CO N T E N T
RAW MATERIALS
Dr. Straetmans is Launching a Range of
Natural Ingredients . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Celligent® and DEFENSIL® Protect Epidermal Stem Cells 66
A New Ecological Range of Cleaning Products . . . . . . . . 66
Kelisoft™ – A Hair Growth Minimiser for Beards and
Underarms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
40 Cognis Products Certified with NaTrue Label . . . . . . . 67
To Regulate the Level of Tolerance and the Long-Term
Balance of the Skin – Soothing . . . . . . . . . . . . . . . . . . . 67
Scientific Data on Skin Penetration Behavior of
PARSOL® SLX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
DURAGREEN™ 135 Polymer – First Zinc-free Technology
for Household Floor Care Market . . . . . . . . . . . . . . . . . . 68
Formulation:
LIGHTEN UP –
Skin Color Refining Wipes . . . . . . . . . . . . . . . . . . . . . .8
EVENT
PCHi Opens for Pre-Registration
22-24 February 2011, Shenzhen Convention &
Exhibition Centre . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
COMPANY NEWS
Azelis Appoints Benoit Fritz as Principals Development
Director . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
Formulae and Dossiers Online on Internet . . . . . . . . .
Gattefossé Announces New GMP Certification for its
. . 73
Cosmetic Operations . . . . . . . . . . . . . . . . . . . . . . . . . . 73
Shastry Appointed Technology Manager for Arizona
Chemical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Aqua Bio Technology: Sales Agreement . . . . . . . . . . . . . 74
Authors’ Guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Impressum / Advertisers Index . . . . . . . . . . . . . . . . . . . 76
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CO S M ET I C S
TA P E ST R I P P I N G

J. Lademann, H.-J. Weigmann, S. Schanzer, M. Meinke, W. Sterry, A. Patzelt*

Analysis of the Penetration of Topically Applied

Substances into the Human Skin

 Introduction
The investigation of the penetration and
storage of topically applied substances
in the human skin is of principal interest
for the development, optimization and
evaluation of topically applied medical
drugs and cosmetic products (1-3).
The number of methods available for the
qualitative and quantitative analysis of
penetration processes under in vivo con-
ditions is small. The topical application
of radio-labelled substances and subse-
quent analysis of urine and blood sam-
the tape stripping procedure (10, 11), layer by layer. The removed tape strips
which has been frequently used for contain a certain amount of corneocytes
decades. After application and penetra- and a certain amount of the topically ap-
tion of topically applied substances, an plied substance, which had penetrated
adhesive film is pressed onto the skin and previously. The concentration of the top-
removed afterwards. Repeating this pro- ically applied substance on each tape
cedure successively on the same skin strip can be determined using classical
area, the stratum corneum is removed analytical methods (12).
Abstract
he knowledge of the penetration and storage of topically applied sub-

ples of the volunteers (4) represents one
method, which is, however, strongly lim-
ited, because of safety standards (5).
Also the removal of biopsies from pre-
treated skin and the analysis of the dis-
tribution of the topically applied sub-
stances in histological sections (6) repre-
sents a highly invasive procedure, which
is not applicable for ethical reasons for
dermatopharmacokinetical investigations,
which require the removal of a series of
biopsies at different time points. A promis-
ing method for dermatopharmacokinet-
ic investigations is the application of mi-
crodialysis (7). In this case, small tubes
consisting of porous membranes are po-
sitioned in the skin. The tubes are con-
tinuously floated by a receptor fluid.
Penetrating through the skin barrier, the
topically applied substances reach the
membranes and penetrate into the re-
ceptor fluid, where they can be detect-
ed. Nevertheless, although microdialysis
is an invasive method, it has obtained an
increased application during the past
few years (8, 9).
A very easy and non-invasive method for
the analysis of the penetration process is
T
stances in the skin is essential for the development, optimization and
evaluation of drugs and cosmetic products.
Tape stripping is a well-known method deployed for the analysis of pene-
tration and storage processes in the skin. Therefore, adhesive films are
placed consecutively on the same skin area to remove the stratum
corneum, layer by layer, after topical application and penetration of sub-
stances. In the past, the penetration depth of a substance was estimated
by the number of tape strips needed to remove the penetrated substance
from the skin. However, the amount of corneocytes removed with one
tape strip can be influenced by topically applied formulations.
Consequently, a comparison of the penetration depths of different formula-
tions is not possible. Therefore, different methods were proposed to deter-
mine the amount of removed corneocytes on the tape strips, quantitative-
ly, in order to identify the thickness of the stratum corneum and the real
penetration depth of the topically applied substance in relation to each
tape strip. As a result, standardized penetration profiles can be assessed,
which represent a cut through the SC and demonstrate the distribution of
the topically applied substances in the horny layer. The application of this
method for the analysis of the penetration and storage of different topical-
ly applied substances is demonstrated in this paper.

2 SOFW-Journal | 136 | 11-2010

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CO S M ET I C S
TA P E ST R I P P I N G
 Problems of the Tape Stripping
Method
Although tape stripping represents a
very simple and non-invasive method, it
provides some problematic aspects:
The tape stripping procedure is a method
to determine the penetration of topical-
ly applied substances only into the stra-
tum corneum (SC). The viable epidermis
is not accessible to this procedure as the
adhesion of the living cells below the
horny layer is too strong to be removed
by tape stripping (13, 14). Nevertheless,
this does not represent a real limitation
for this method. As the SC represents the
barrier of our organism, only the pene-
tration process inside and through this
barrier is of real interest. After crossing
the stratum corneum, the topically ap-
plied substances are immediately uptak-
en by the living cells or by the blood or
lymph flow (15).
In the traditional method of tape strip-
ping, the number of tape strips is used to
determine the penetration depth of the
topically applied substances (16). How-
ever, this method does not allow a cor-
rect determination of the real penetra-
tion depth as the number of corneocytes
removed with one tape strip can vary
significantly depending on the topically
applied formulation (14). The application
of fatty or oily formulations on the skin
reduces the adhesive forces of the tapes
in comparison to ethanol formulations.
As a consequence, penetration profiles of
different formulations, although removed
from the same volunteer are not compa-
rable, as the tape strips are removed from
different depths of the SC (14).
In order to overcome this problem, dif-
ferent methods have been proposed to
determine not only the amount of topi-
cally applied substances, but also the
amount of removed corneocytes on the
tape strips.
 Methods for the Qualitative and
Quantitative Determination of the
Amount of Corneocytes Removed
by Each Tape Strip
Determination of the weight
In principle, it is possible to determine
the amount of SC on the removed tape
4 SOFW-Journal | 136 | 11-2010
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strips by analyzing the weight of the
tapes before and after their application
on the skin (17). However, this method
can be applied only on the untreated
skin, as tape stripping on pre-treated
skin would lead to an additional removal
of the topically applied substance which
would falsify the weighing results.
Determination of the protein absorption
The protein absorption can be used for
the characterization of the amount of
corneocytes removed with the tape strips
(18). The UV-absorption band at 280 nm
is almost superposed by the strong ab-
sorption band of the topically applied
formulation, so that it cannot be used
for the quantitative determination (17).
In contrast to the UV-band, the IR-ab-
sorption is well suited for the determi-
nation of the amount of corneocytes on
the tape strips. A commercial device based
on this principle has recently been intro-
duced by Voegeli, et al (19).
Determination of the pseudo-absorption
Additionally, also the determination of
the pseudo-absorption is possible to
quantify the amount of corneocytes re-
moved. The absorption spectra of the
first tape strip removed from an un-
treated skin area and an area treated
with a commercial sunscreen are com-
pared in Fig. 1. The UV-absorption bands
-Fig. 1 Absorption spectra of the first tape strip removed from untreated skin and
from skin treated with a commercial sunscreen
of the filter substances can be clearly
recognized in comparison to the untreat-
ed skin. At wavelengths < 430 nm, the
absorption of both spectra are identical.
This value is determined by the pseudo-
absorption of corneocytes on the re-
moved tape strips (20). The pseudo-ab-
sorption is a superposition of absorption,
reflection and scattering of the corneo-
cytes. This value is a measure of the
amount of SC on the removed tape strips,
which can easily be determined by spec-
troscopic measurements.
Transepidermal water loss (TEWL)
In principle, it is also possible to deter-
mine the quantity of the corneocytes on
the tape strips by TEWL measurements
(21). The TEWL value increases with each
removed tape strip, as the skin barrier is
damaged by the tape stripping proce-
dure (13, 21). The application of this
method is limited as it can only be used
on untreated skin, as TEWL values can be
disturbed by topically applied sub-
stances (14).
 Penetration Profiles
The most promising methods for the de-
termination of the amount of corneo-
cytes on the removed tape strips are the
analysis of the IR-protein absorption and
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CO S M ET I C S
TA P E ST R I P P I N G

of the pseudo-absorption, as described

above. If the SC is removed completely
by tape stripping, a complete horny lay-
er profile can be calculated by adding
the single amounts of corneocytes on
each tape strip.
A penetration profile can be calculated
by correlating the amounts of topically
applied substances of each single tape
strip to the corresponding position in the
horny layer profile. A typical example of
such a penetration profile is shown for a
sunscreen formulation in Fig. 2. The dis-
tance between the horizontal lines cor-
responds to the amount of corneocytes
removed with the corresponding tape
strips. The upper line represents the skin
surface; the lower line demonstrates the
boundary between the SC and the living -Fig. 2 Penetration profile of a sunscreen
tissue (stratum granulosum). The dis-

SOFW-Journal | 136 | 11-2010 5

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CO S M ET I C S
TA P E ST R I P P I N G

tances of the horizontal lines decrease

with increasing tape numbers. This re-
flects the well-known fact that less cor-
neocytes can be removed from deeper
parts of the horny layer, because of the
reduced desquamation. Penetration pro-
files represent a non-invasive cut through
the SC, considering the distribution of
topically applied substances in the SC at
the time the tape strips were removed.
As can be seen from Fig. 2, the topically
applied sunscreen is located only in the
upper layers of the stratum corneum
protecting the skin surface.

 Penetration Studies of Topically

Applied Drugs and Cosmetics
-Fig. 3 Penetration profile of a barrier cream containing microparticles

A typical penetration profile of a sun-

screen is demonstrated in Fig. 2. In com- 4a
parison to a typical penetration profile
of a sunscreen formulation (Fig. 2), a
penetration profile of microparticles at a
size of 5 µm applied in an o/w emulsion
is presented in Fig. 3. The microparticles
represent one component of the barrier
cream and are supposed to form a pro-
tection film on the skin surface (22),
which can be read off the penetration
profile (Fig. 3). The microparticles are lo-
cated only on the skin surface as they
were almost completely removed from
the skin with the first tape strip. The mi-
croparticles detected on the second and
third tape strips cannot be considered as
penetrated, but they were located in the 4b
furrows and wrinkles of the skin. In the
demonstrated case, the microparticles
were completely removed by four tape
strips (22).
In contrast, the penetration profiles of
topically applied medical drugs, which
are supposed to pass the skin barrier,
show different penetration behaviours.
In Fig. 4, penetration profiles of the steroid
Clobetasol in different formulations are
demonstrated. In both cases, the active
substance was detectable up to the bound-
ary to the living cells, which means a
penetration through the skin barrier in
the direction to the living cells. It is well-
known that the penetration process is
strongly influenced by the formulation,
in which the drug is applied. This effect -Fig. 4 Penetration profiles of steroids (Clobetasol) in two different formulations
can be clearly seen from Figs. 4a and b. (4a: Formulation I - Temovate emollient cream; 4b: Formulation II – Temovate cream)
In the first case (Formulation I), the drug

6 SOFW-Journal | 136 | 11-2010

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CO S M ET I C S
TA P E ST R I P P I N G

is practically stored on the skin surface

and in the upper layers of the SC (Fig.
4a). In the second case (Formulation II),
a high concentration of the steroid was
also detected in deeper parts of the SC
(Fig. 4b). Therefore, the penetration of
Clobetasol applied in Formulation II is
much better than in the case of Formu-
lation I. This corresponds to the differ-
ences in the intensity of the biological
response of the skin in form of vasocon-
striction after application of both formu-
lations.
The tape stripping procedure can also be
used for the analysis of dermatophar-
macokinetics of topically applied sub-
stances. In this case, the penetration pro-
files have to be determined at different
time points after application. The changes
in the distribution of the substances in-
side the SC can be well detected; where-
by qualitative and quantitative informa-
tion on the penetration kinetics of the
topically applied substances can be ob-
tained.

 Conclusions

Summarizing the results presented in

this paper, it can be stated that the tape
stripping method is an easy, fast and
cheap method for the analysis of the
penetration of topically applied drugs
and cosmetic products into and through
the skin barrier.
It is essential for the application of the
tape stripping procedure that the amount
of substances determined on the re-
moved tape strips is related to the posi-
tion of the horny layer profile, from
where the corresponding tape strips were
removed. Only in this case, a standard-
ized qualitative and quantitative analy-
sis of the penetration profile becomes
possible.

Acknowledgements

The authors would like to thank the

Foundation »Skin Physiology« of the
Donor Association for German Science
and Humanities for financial support.

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TA P E ST R I P P I N G

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* Authors:
J. Lademann, H.-J. Weigmann
S. Schanzer, M. Meinke, W. Sterry
A. Patzelt
* Correspondence address:
Prof. Dr. Juergen Lademann
Charité – Universitätsmedizin Berlin
Department of Dermatology and Aller-
gology
Center of Experimental and Applied
Cutaneous Physiology
Charitéplatz 1
10117 Berlin
Germany
Email: juergen.lademann@charite.de


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CO S M ET I C S
S K I N I M MU N I T Y

G. Dell’Acqua*, K. Schweikert, G. Calloni

Stimulating and Protecting Skin Immunity to

Decrease UV-Induced Skin Erythema

 Introduction With a reduced immune defense, the skin with associated inflammation and pro-
would be less reactive and unbalanced vide therefore a long term skin soothing.
Skin is permanently exposed to stress with the need of regaining its proper In order to stimulate innate immunity
from the external environment. In order healthy balance and reactivity. and early defenses we have selected in-
to defend itself and to increase its re- The approach is then to stimulate innate gredients that have been described in
pairing capacity, skin possesses mole- immunity markers and Langerhans cells the scientific literature for their poten-
cules that are part of the innate immu- with the goal to re-balance an under-re- tial in soothing skin irritation through
nity system. These molecules are ex- acting skin. Doing so, we believe we can modulation of the immune system. A
pressed by the keratinocytes and also reduce upcoming external aggressions bioactive complex was formulated, con-
present on Langerhans cells. They are
highly conserved through evolution and
represent the first line of defense against
foreign antigens and environmental stress.
Although during the early response these
molecules act locally, they may trigger
eventually a more systemic immune re-
sponse if the aggression can not be re-
solved rapidly. These molecules also called Abstract
innate immunity markers can be consid-
kin defense and reactivity involve production by keratinocytes of in-

ered, together with the Langerhans cells,
the skin immune sentinels (1) making
sure that a pro-inflammatory aggression
is detected and controlled (1-4). Among
the skin immunity markers we can list
anti-microbial peptides like cathelicidins
and defensins that directly kill invasive
microbes (5, 6); heme oxygenase 1 (HO-1),
involved as an anti-oxidant and wound
healing agent (7, 8); S100 proteins, with
both anti-microbial (9) and skin barrier
properties (10); and Toll like receptor-2
with signaling function (10) but also im-
portant in anti-microbial defense (11,
12).
Unfortunately, external stress and in
particular UV light can significantly re-
duce the innate immunity markers and
the Langerhans cells (13-16), depriving
the skin from its basic defenses and in-
creasing the possibility of antigen-trig-
gered inflammation.
S
nate immunity proteins. These proteins, also expressed on skin
Langerhans cells (the immunity sentinels), help the skin reacting to
environmental aggressions and repairing damage. A complex of natural in-
gredients was formulated to stimulate and to protect skin’s immunity. A
combination of middle weight polysaccharides from Tamarindus and glyco-
side Stevioside (trade name: Unisooth ST-32) was able to stimulate in hu-
man keratinocytes, markers of innate immunity such as Defensin beta 4
(+127%) and Heme Oxygenase 1 (+51%) and to protect Langerhans cells
from UV-induced down regulation in human skin (63% protection). Further-
more, the complex formulated at 3%, reduced significantly UV-induced
erythema in human volunteers (n=25) by 59% and Trans Epidermal Water
Loss (TEWL) by 68% when compared to a placebo. In conclusion, it is possi-
ble to significantly prevent and reduce UV-induced irritation by boosting
skin’s immunity with a combination of natural ingredients. The complex is
suitable for day, 24 hours and sensitive skin application products.

10 SOFW-Journal | 136 | 11-2010


taining a balanced blend of Tamarindus
middle weight polysaccharides extract-
ed from the plant’s seeds combined with
the glycoside Stevioside, extracted from
the Stevia leaves (trade name: Unisooth
ST-32). Tamarindus Indica has been as-
sociated to anti-inflammatory proper-
ties in traditional medicine (17) and
more recently its polysaccharides have
shown soothing characteristics, being
often compared to Aloe oligosaccharides
for their efficacy (18, 19). Tamarindus
polysaccharides have also shown the ca-
pacity to protect against UV-induced
damage (20), suggesting its utilization as
a UV protector.
Stevioside is a well know sweetener in
the food industry, however its healthy
benefits have been recently evidenced
(21). In particular, its capacity as an im-
mune modulator and anti-inflammatory
characteristics have been studied (22,
23).
The hypothesis was then to test this
complex to stimulate, in human ker-
atinocytes, the innate immunity markers
responsible for skin defense and repair
and to protect, in human skin explants,
Langerhans cells from UV-induced de-
pletion (Fig. 1). Finally, the complex for-
mulated at 3% in a water based gel was
applied on human volunteers for reduc-
ing UV-induced erythema and trans-
epidermal water loss (TEWL).
 Materials and Methods
Normal human epidermal keratinocytes
(NHEK) culture, treatment and analysis
for innate immunity markers
NHEK at 3rd passage were seeded in 24-
well plates in culture medium and incu-
bated until confluence. The medium was
then replaced by assay medium contain-
ing or not Stevioside at concentrations
of 0.2%, 0.03%, and 0.001% or Tamarindus
polysaccharides at concentration of 0.01%
and left for 24 hours. All experimental
conditions were performed in triplicate
(n=3). At the end of the incubation, the
cells were washed in phosphate buffered
saline (PBS) solution and extracted for
mRNA; mRNA was reverse-transcribed
and genes for Cathelicidin, Defensin be-
ta 4, Heme Oxygenase 1 (HO-1), S100 A7,
Toll-like receptor-1, were analysed.
SOFW-Journal | 136 | 11-2010
< Content
CO S M ET I C S
S K I N I M MU N I T Y
-Fig. 1 Unisooth ST-32 protects and sooths skin epidermis from external aggres-
sions. It increases synthesis of innate immunity proteins in keratinocytes and pro-
tects Langerhans cells to boost skin defenses and to stimulate skin repair for an
overall long term soothing effect. LC: Langerhans cell
Skin explants culture, treatment and Induction and measurement of Erythe-
analysis for Langerhans cells ma and Trans-Epidermal Water Loss
Skin explants from plastic surgery were (TEWL) in human volunteers (double
treated topically with 20 mg/cm2 of blind study)
Unisooth ST-32 (3% in water) or a con- Skin erythema was induced on human
trol reference (SPF30 solar cream), and volunteers (n = 25) by exposure to UV
incubated for 24 hours. Explants were then light (UVA + UVB, solar spectrum). On
irradiated with a total dose of 1500 mJ/ each volunteer, the MED was determined
cm2 UVB (SOL500 Sun Simulator with H2 by applying a pattern of radiation con-
filter). Non irradiated controls were kept sisting in 6 doses of UV radiation. The ra-
in the dark. After irradiation, the skin ex- diation pattern was chosen based on the
plants were again treated with Unisooth subject phototype. Once each individual
ST-32 or the control reference and incu- MED was established, an erythematic re-
bated for additional 24 hours. At the end action was induced in 9 skin areas of the
of the incubation, punches were per- volunteers back by using an UV irradia-
formed on each skin explants, tissue tion corresponding to 1.25x MED. 6 hours
sectioned and incubated with an anti- before irradiation (T0), 2 mg/cm2 prod-
CD1a-FITC antibody specifically recog- uct was applied on the back of the
nizing Langerhans cells (LC). Observa- volunteers. 3 areas were treated with a
tions were performed in epifluorescence placebo gel, 3 areas were treated with
with a Nikon E400 microscope and fluo- a water based gel containing 3% of
rescent LC were counted (n = 45/condi- Unisooth ST-32 and 3 areas were left un-
tion). Digital images were recorded with treated (irradiated CT). Skin redness (evalu-
a Nikon DXM 1200F camera and Lucia 4.8 ated by Erythema index) and TEWL were
software. measured 24 hours and 48 hours from T0.
Percentage of protection was calculated Comparison was made between all sam-
according to the following formula: ples and to the untreated areas. Erythe-
Positive Control (with UV) – Treatment + UV
Protection (%) = X 100
Positive Control (with UV) – Negative Control (without UV)
11
 < Content
CO S M ET I C S
S K I N I M MU N I T Y

ma index was measured using a Mexa-

meter MX 18 (Courage+Khazaka, elec-
tronic GmbH), while TEWL was measured
using a Tewameter 300 (Courage+Kha-
zaka, electronic GmbH). Data were ana-
lyzed and expressed as % variation vs T0.
Statistical significance was also calculat-
ed.

 Statistical Analysis

The inter-group comparisons were per-

formed by Student’s T test.

-Fig. 2 Stevioside stimulates innate immunity markers mRNA synthesis in normal

 Results human epidermal keratinocytes

Stimulation of innate immunity mark-

ers in NHEK duction in UV-induced skin erythema of This decrease was statistically significant
Treatment of NHEK with 0.01% Tama- 48.7% and 58.8% when compared to a (p<0.001 vs placebo, Student’s T test). In
rindus polysaccharides did not bring any placebo treatment, after 24 hours and 48 the same volunteers, UV-induced skin
differences to innate immunity markers hours from UV-irradiation, respectively. trans-epidermal water loss (TEWL) was
(not shown), while treatment of NHEK
with different concentrations of Stevio-
side, increased the transcription of in-
nate immunity markers (Cathelicidin, De-
fensin beta 4, S100 A7, Toll-like receptor
2 and Heme oxygenase 1) as shown in
Fig. 2. This increase was dose dependent.
In particular, Defensin beta 4 was strong-
ly stimulated even at concentration of
0.03%. -Fig. 3 Treatment of human skin explants with Unisooth ST-32 at 3% provides a
63% protection from UVB-induced Langerhans cells depletion. CT - UVB = not
Protection of Langerhans cells in human
treated skin control; CT + UVB = treated skin control (1500 mJ/cm2 UVB). Langer-
skin explants
hans cells are stained in green
In irradiated skin explants, the number
of Langerhans cells in epidermis was
significant lower compared to control
non-irradiated explants. Treatment with UV-induced erythema
Unisooth ST-32 at 3% significantly
protected from UV-induced depletion
Langerhans cells. The protection was
63% (p<0.001, Student’s T test). Im-
munofluorescence pictures are shown in
Fig. 3. Topical treatment with control
reference sun cream SPF30 protected the
explants against UV-induced Langer-
hans cells depletion by 67% (data not
shown).

Reduction of UV-induced erythema

and Trans-epidermal water loss (TEWL)
in human volunteers
As shown in Fig. 4, pre-treatment of hu-
man volunteers with Unisooth ST-32 at
3% in a water based gel, induced a re-
-Fig. 4 Pre-treatment with a gel containing Unisooth ST-32 at 3.0% reduces UV-in-
duced erythema on human volunteers (n = 25). UV irradiation = 1.25xMED.
*Significant data vs Placebo

12 SOFW-Journal | 136 | 11-2010

 < Content

Soothing
A new approach to regulate the
level of tolerance and the long-term
balance of the skin

Unisooth ST-32 stimulates the skin defense

· by boosting the immunity mechanism of the skin
· by protecting the Langerhans cells
Unisooth PN-47 modulates the skin reactivity
· by using powerful JAK/STAT inhibitors
Unisooth EG-28 modulates the skin reactivity
· by using powerful NF-kB inhibitors
Unisooth PN-47 and EG-28 combined,
they block the keratinocyte activation loop.
Induchem
www.induchem.com Innovative • Collaborative • Reliable

CO S M ET I C S
S K I N I M MU N I T Y
reduced by the gel containing Unisooth
ST-32 at 3%. The decrease was of 68.3%
and 68.4% when compared to a placebo
treatment, after 24 hours and 48 hours
from UV-irradiation, respectively (Fig. 5).
This decrease was statistically significant
(p<0.001 vs placebo, Student’s T test).
The reduction of both the erythema and
the TEWL suggests that Unisooth ST-32
may not only have a soothing but also a
healing effect.
 Conclusion
Unisooth ST-32 stimulates synthesis of
innate immunity markers and protects
Langerhans cells. These effects suggest
that Unisooth ST-32 can have immuno-
stimulating and protecting properties.
Since UV light and stress induce immuno
suppression and reduced response to en-
vironmental aggressions (13-16), it is
necessary to boost skin defenses mech-
anisms linked to innate immunity mark-
ers such as the anti-microbial peptides,
repairing markers such as S100A7 and
HO-1 and to protect Langerhans cells.
The observed stimulation by Stevioside
and Tamarindus polysaccharides of these
mechanisms confirm the capacity of
these ingredients to stimulate a mecha-
nism of protection and repair that can be
associated to a long term soothing. The
effect on Langerhans cells was almost as
good as an SPF30 cream used as a con-
trol reference (63% vs 67%, data not
shown).
When Unisooth ST-32 was incorporated
in a water based gel at concentration of
3% and applied topically on human vol-
unteers before and after UV irradiation,
it was able to significantly reduce the
UV-induced erythema. The effect was
visible already after 24 hours from UV ir-
radiation and increased after 48 hours.
Interestingly, Unisooth ST-32 was also
able to dramatically decrease the trans-
epidermal water loss (TEWL) induced by
the UV irradiation. This effect was statis-
tically significant and already impressive
after 24 hours, while not increasing af-
ter 48 hours.
These data in vivo support the utilization
of Unisooth ST-32 immuno-modulatory
characteristics to reduce a UV-induced
irritation.
14
< Content
-Fig. 5 Pre-treatment with a gel containing Unisooth ST-32 at 3.0% reduces UV-in-
duced TEWL on human volunteers (n = 25). UV irradiation = 1.25xMED.
*Significant data vs Placebo
In conclusion, we have demonstrated
that Unisooth ST-32, by stimulating
mechanisms linked to skin protection
and repair, can indeed act as a long term
soothing agent. Moreover its effect in
reducing the TEWL can also suggest a
role as a healing agent in restoring the
damaged skin barrier.
Unisooth ST-32 can be incorporated in
products for daily protection and repair
to provide a long term soothing and de-
crease the risk of environmental aggres-
sions. Day and 24 hours products are
certainly a suggested application, but
Unisooth ST-32 can also be recommend-
ed in products for sensitive skin individ-
uals where a constant soothing is need-
ed. Recommended usage levels would be
between 1.0% and 3.0%.
Unisooth ST-32 has been tested for skin
tolerance, mutagenicity and biodegrad-
ability, and has provided an excellent
safety profile.
References
(1) Nestle FO, Di Meglio P, Qin JZ, Nickoloff BJ
(2009) Skin immune sentinels in health and
disease. Nat Rev Immunol 9:679-91
(2) Goodarzi H, Trowbridge J, Gallo RL (2007) In-
nate immunity: a cutaneous perspective. Clin
Rev Allergy Immunol 33:15-26
(3) Merad M, Ginhoux F, Collin M (2008) Origin,
homeostasis and function of Langerhans cells
and other langerin-expressing dendritic cells.
Nat Rev Immunol 8:935-47
(4) Clausen BE, Kel JM (2010) Langerhans cells:
critical regulators of skin immunity? Immunol
Cell Biol 88:351-60
(5) Braff MH, Gallo RL (2006) Antimicrobial pep-
tides: an essential component of the skin de-
fensive barrier. Curr Top Microbiol Immunol
306:91-110
(6) Niyonsaba F, Nagaoka I, Ogawa H (2006) Hu-
man defensins and cathelicidins in the skin:
beyond direct antimicrobial properties. Crit
Rev Immunol 26:545-76
(7) Alcaraz MJ, Fernández P, Guillén MI (2003) An-
ti-inflammatory actions of the heme oxyge-
nase-1 pathway. Curr Pharm Des 9:2541-51
(8) Hanselmann C, Mauch C, Werner S (2001)
Haem oxygenase-1: a novel player in cuta-
neous wound repair and psoriasis? Biochem J
353:459-66
(9) Eckert RL, Broome AM, Ruse M, Robinson N,
Ryan D, Lee K (2004) S100 proteins in the epi-
dermis. J Invest Dermatol 123:23-33
(10) Köllisch G, Kalali BN, Voelcker V, Wallich R,
Behrendt H, Ring J, Bauer S, Jakob T, Mempel
M, Ollert M (2005) Various members of the
Toll-like receptor family contribute to the in-
nate immune response of human epidermal
keratinocytes. Immunology 114:531-41
(11) Gläser R, Harder J, Lange H, Bartels J,
Christophers E, Schröder JM (2005) Antimi-
crobial psoriasin (S100A7) protects human
skin from Escherichia coli infection. Nat Im-
munol 6:57-64
(12) Lai Y, Cogen AL, Radek KA, Park HJ, Macleod
DT, Leichtle A, Ryan AF, Di Nardo A, Gallo RL
(2010) Activation of TLR2 by a small molecule
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S K I N I M MU N I T Y

produced by Staphylococcus epidermidis in-
creases antimicrobial defense against bacte-
rial skin infections. J Invest Dermatol 130:
2211-21
(13) Clydesdale GJ, Dandie GW, Muller HK (2001)
Ultraviolet light induced injury: immunolog-
ical and inflammatory effects. Immunol Cell
Biol 79:547-68
(14) Norval M (2006) The mechanisms and conse-
quences of ultraviolet-induced immunosup-
pression. Prog Biophys Mol Biol 92:108-18
(15) Grewe M (2001) Chronological ageing and
photoageing of dendritic cells. Clin Exp Der-
matol 26:608-12
(16) Aberer W, Schuler G, Stingl G, Hönigsmann H,
Wolff K (1981) Ultraviolet light depletes sur-
face markers of Langerhans cells. J Invest Der-
matol 76:202-10
(17) Rimbau V, Cerdan C, Vila R, Iglesias J (1999)
Antiinflammatory activity of some extracts
from plants used in the traditional medicine
of north-African countries (II). Phytother Res
13:128-32
(18) Strickland FM, Kuchel JM, Halliday GM (2004) (23) Boonkaewwan C, Toskulkao C, Vongsakul M
Natural products as aids for protecting the (2006) Anti-Inflammatory and immunomodu-
skin's immune system against UV damage. latory activities of Stevioside and its metabo-
Cutis 74:24-8 lite Steviol on THP-1 cells. J Agric Food Chem
54:785-9
(19) Strickland FM, Darvill A, Albersheim P, Eber-
hard S, Pauly M, Pelley RP (1999) Inhibition
of UV-induced immune suppression and in-
terleukin-10 production by plant oligosac-
charides and polysaccharides. Photochem Authors:
Photobiol 69:141-7 Giorgio Dell’Acqua, Kuno Schweikert
Giuseppe Calloni
(20) Raimondi L, Lodovici M, Guglielmi F, Banchel-
li G, Ciuffi M, Boldrini E, Pirisino R (2003) The
polysaccharide from Tamarindus indica (TS- * Correspondence address:
polysaccharide) protects cultured corneal- Giorgio Dell’Acqua
derived cells (SIRC cells) from ultraviolet rays. Induchem AG
J Pharm Pharmacol 55:333-8 Industriestraße 8a
(21) Chatsudthipong V, Muanprasat C (2009) Ste- 8604 Volketswil
vioside and related compounds: therapeutic Switzerland
benefits beyond sweetness. Pharmacol Ther Email:
121:41-54 giorgio.dellacqua@induchem.com
(22) Sehar I, Kaul A, Bani S, Pal HC, Saxena AK
(2008) Immune up regulatory response of a

non-caloric natural sweetener, stevioside.
Chem Biol Interact 173:115-21

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SOFW-Journal | 136 | 11-2010 15


CO S M ET I C S
SENSITIVE SKIN
J.W. Fluhr*, R. Darlenski**
Development of Skin Models –
Sensitive Skin
 Introduction
Sensitive skin is a term referred to ab-
normal sensory skin reactivity. The con-
dition, also known as the »sensitive skin
syndrome«, gained broad popularity not
only among the scientific community
but also in the general population as
well. Around 50% of the adult popula-
tion report unpleasant facial sensations
such as stinging, prickling, burning, itch-
ing and pain (1).
Sensitive skin poses a challenge not on-
ly to the clinician but also to the cos-
metic industry. There is no golden stan-
dard for testing the efficacy of cosmet-
ic products for sensitive skin. Therefore,
the creation and validation of a skin
model for sensitive skin is a »hot topic«
in dermatological and cosmetological
research. Recent ethical considerations
resulted in the prohibition of animal test-
ing with the 7th Amendment to Cosmet-
ic Directive of EU (76/768/EEC). A trend
towards the use of in vivo human and/or
in vitro methods is observable, in accor-
dance with the EU directive. In the case
of »sensitive skin« there is no established
in vitro assay and no model can be effi-
ciently applied in this condiciton as it is
mainly characterized by subjective symp-
toms. Therefore, testing sensitive skin
products is only applicable in human
studies.
The need for quantification and objec-
tivity with regard to clinical endpoints
lead to the introduction of a number of
tests and methods aiming to character-
ize different aspects of sensitive skin
such as the sensory and objective irrita-
tion tests, and the assessment of skin
functional properties.
16
< Content
 What is Sensitive Skin? present, although no visible signs of irri-
tation are observable in most of the cas-
Historically, the term sensitive skin was es. Different terms have been allied to
introduced in the 1970s and since then this entity such as sensory, subjective,
it belongs to the group of so called »in- subclinical, and invisible irritation as well
visible dermatoses« (2) due to the lack of as sensitive skin syndrome and cosmetic
objective clinical findings in most of the intolerance syndrome (3). Sensitive skin
cases. There is no uniform agreement on syndrome is referred to an abnormal sen-
the definition of »sensitive skin«. Subjec- sory reaction to skin care products and
tive hyperreactivity to a number of en- cosmetics (4). Others add to the trigger-
vironmental factors is characteristic for ing agents environmental factors such as
subjects with sensitive skin. Itching, burn- cold, heat, high humidity, and dry air
ing, stinging, and/or tight sensation are flow (5).
Abstract
ifferent in vivo models for dry skin, disrupted skin conditions, sensi-
D
tive skin and aging skin have been elaborated in recent years. Vari-
ous studies based on non-invasive biophysical methods have noted
important differences in the skin of »stingers« compared to »non-stingers«:
A relationship between sensory irritation and epidermal barrier function.
Specific knowledge regarding skin physiology is now available (Fig. 1).
Higher baseline transepidermal water loss (TEWL) values were reported in
stingers compared to non-stingers. These subjects also showed a more ex-
aggerated increase in TEWL at sites of experimentally induced acute irritant
dermatitis. The stratum corneum of stingers seems to be less hydrated, as
evidenced by lower electrical capacitance values. They might present high-
er baseline surface pH values and lower sebum content. Stingers also ap-
pear to have a more reactive cutaneous vascularisation. This article de-
scribes new models and their non-invasive evaluation. Current knowledge
on biophysical assessment in identifying and characterising sensitive skin
will be further discussed.
SOFW-Journal | 136 | 11-2010
 < Content
CO S M ET I C S
SENSITIVE SKIN

It was suggested that »sensitive skin« in-

volves four categories: 1. Individuals with
obvious skin disease; 2. Subjects with
subclinical (mild) or atypical clinical signs
of disease; 3. Individuals with history of
past skin disorder; 4. Subjects who do not
fit into one of the above three categories
and appear to be »normal« (6).

 The Burden of Sensitive Skin

The estimation of the exact population

prevalence of sensitive skin is hindered
because there is no widely accepted di-
agnostic approach to the condition. Dis-
crepancies in the epidemiological data
exist. Approximately 40% of the popu-
lation considers themselves to possess
-Fig. 1 Mechanisms of the altered skin functional properties in sensitive skin the characteristics of sensitive skin (7).
(LAST – lactic acid stringing test; TEWL – transepidmeral water loss; SC – Stratum Population’s demographics influence the
corneum) prevalence of the condition. A mail sur-
vey in UK disclosed prevalence of self-re-

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SOFW-Journal | 136 | 11-2010 17


CO S M ET I C S
SENSITIVE SKIN
ported sensitive skin in 51.5% of women
and in 38.2% of men (8). 10% of women
and 5.8% of men described themselves
as having very sensitive skin. In USA,
52% of women between 18 and 45 years
of age declared sensitive facial skin with
no significant difference between ethnic
groups (9). The female predominance was
confirmed by other studies based on the
subjects’ perception (10, 11).
According to a population-based survey
42% believed that they had sensitive
skin with female predominance (59%)
against males (44%) (11). The probabili-
ty of having a concomitant skin disorder
is higher in subjects claiming very sensi-
tive skin (28%) than in those with sensi-
tive skin (14%), not very sensitive skin
(7%), and in those with normal skin (2%).
This is consistent with the concept that
existent dermatologic diseases, such as
rosacea and different type of eczema can
be conductive to sensitive skin condition
(7, 8, 12). A summary of the skin diseases
coexistent with sensitive skin syndrome
is presented in Table 1.
Sensitive skin is accompanied by wors-
ened quality of life with affection of the
psychological dimension rather than a
predominant disturbed physical compo-
nent (10, 11). However, it cannot be con-
cluded whether the skin condition is the
direct cause for these abnormalities. Ad-
ditional social and environmental fac-
tors such as climate, ethnicity, educa-
tion, occupation, cosmetic habits, and
media influence affect the self-percep-
tiveness of sensitive skin (10, 13).
 Pathophysiological Grounds for
Developing Sensitive Skin Model
The development of a successful sensi-
tive skin model relies on the intimate
pathological abnormalities in this phe-
nomenon. Although the exact aetiology
of the condition is still to be elucidated,
functional cutaneous abnormalities of
the sensitive skin subjects have been de-
scribed. Defective epidermal barrier func-
tion, increased transcutaneous penetra-
tion, hyperreactivity of the blood vessels,
and variation in the number and the
stimulation response in the neurosenso-
ry fibers are implicated in altered skin
sensitivity.
18
< Content
Desease Group Skin Diseases with Altered Skin Sensitivity
Disorders with altered Allergic contact dermatitis
skin reactivity Irritant contact dermatitis
Atopic dermatitis
Eczema (including nummular eczema and
lichen simplex chronicus)
Seborrheic dermatitis
Immunologic and non-immunologic contact
urticaria
Disorders of epidermal Rosacea
appendages and related disorders Acne (including comedonic, papular and
pustular)
Perioral dermatitis
Disorders of persistent Psoriasis
inflammation, cell kinetics,
and differentiation
Table 1 Summary on the skin diseases associated with sensitive skin.
(Adapted from Z.D. Draelos, 2000 (31). The relation of the disorders to a certain
group is based on the classification of the skin diseases presented in the Fitzpatrick’s
Dermatology in General Medicine, 7th edition, 2007 (32))
Noninvasive methods are applied in in- Increased penetration of methyl nicoti-
vestigating the structure and functions nate (MN) was demonstrated by assess-
of sensitive skin subjects. Parameters of ment of erythema in sensitive skin sub-
cutaneous physiology changes (TEWL, SC jects (19). This is consistent with more
hydration, skin surface pH, and skin prominent local vasodilatory effect of
colour), indicative of sensitive skin, can MN as registered by laser-doppler ve-
be detected at low levels prior to clinical locimetry in sensitive skin individuals
disease presentation (14). (19, 21). Thus the increased vascular re-
A trend towards higher TEWL baseline val- activity in sensitive subjects might be re-
ues witnessing impaired epidermal barri- lated to an enhanced penetration of sen-
er function has been revealed (15, 16). sory irritants, thus facilitating the sting-
The hydration of SC as assessed by mea- ing sensation. Confirmation comes from
suring skin capacitance was lower on the clinical trials, as laser treatment of rosacea
cheek of individuals with self-reported destroying small vessels resulted in a de-
sensitive skin (17). A propensity toward crease of the LAST scores (22).
higher LAST scores was characteristic for Acidic skin surface pH is necessary to
these subjects. Dehydration might be the maintain bacteriological, chemical and
basis for subjective sensations after ex- mechanical resistance of the skin, as well
posure to water and soap. Lower base- to sustain the epidermal barrier home-
line capacitance values are observed in ostasis and the SC integrity and cohesion
sensitive skin subjects (15, 16), although (23, 24). In sensitive skin subjects skin
single reports could not demonstrated surface pH is slightly higher or equal to
a difference in SC hydration between that in normal individuals (16-18).
»stingers« and »non-stingers« (18). Mor- A lower level of skin surface lipids as as-
phological studies in subjects with hy- sessed by photometric technique was
perreactive skin showed thinner SC with shown in »stingers« compared to »non-
reduced corneocyte surface area (19) and stingers« under basal conditions. A re-
irregular skin surface microrelief (20). duction in the surface lipids on day 14
SOFW-Journal | 136 | 11-2010
 Editor:
< Content
Deutsche Gesellschaft für
Wissenschaftliche und Angewandte Kosmetik e.V. German Society for Scientific and Applied Cosmetics e.V. (DGK)
Expert Group »Microbiology and Industrial Hygiene«

Industrial Hygiene
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in the Cosmetics Sector
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even this is very fragmented. This book aims to fill the current gaps by bringing
together all the important aspects of industrial hygiene and providing a clear
overview of the current knowledge in this specialist field.
This book has been written from a microbiological and hygienic perspective and
aims to provide practitioners such as plant managers, developers, quality as-
Verlag für chemische Industrie U H. Ziolkowsky GmbH U Augsburg
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dustrial hygiene.

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1 Industrial hygiene – a key part of the 4.9 Taking samples to determine microbiological strain 7.6 Handling semi-finished (bulk) and finished goods
cosmetics manufacturing process 4.10 Disinfecting the system 7.7 Guidance values and control
1.1 Hygiene – part of the quality system 4.11 Common technical and hygienic errors 7.8 Literature
1.2 Implementing hygiene requirements 4.12 Literature
1.3 Training and control measures 8 Pest control and waste disposal
1.4 Monitoring hygiene in relation to external service 5 Room hygiene 8.1 Pest control
providers, suppliers and contract 5.1 Cleaning and disinfection 8.2 Waste disposal
1.5 Literature 5.2 Hygiene schedule (for cleaning and disinfecting 8.3 Literature
rooms subject to the good manufacturing practice
2 Structural requirements requirements) 9 Microbiological controlling
2.1 General building requirements – 5.3 Personnel movements and material flows 9.1 Sampling
room specification sheets 5.4 Ventilation 9.2 Process control (raw materials – packaging –
2.2 Site requirements 5.5 Literature preparation vessel – storage – filling – packing)
2.3 Planning principles 5.6 Sample documents 9.3 Methods
2.4 Building areas 9.4 Alternative methods
2.5 Literature 6 Personnel hygiene
6.1 Integration into the hygiene areas concept 9.5 Organising and evaluating microbiological control
3 System design and System hygiene 6.2 Employee conduct 9.6 Literature
3.1 System design 6.3 Hand hygiene
3.2 System hygiene 6.4 Clothing 10 Hygiene monitoring
3.3 Literature 6.5 Health 10.1 Methods
6.6 Visitors and external personnel 10.2 Organising and implementing hygiene control
4 System design and System hygiene for 6.7 Employee training 10.3 Hygiene control schedules
Water 6.8 Controlling 10.4 Guidance values
4.1 Microbiological quality of the product 6.9 Documentation 10.5 Documentation
water used 6.10 Miscellaneous 10.6 Literature
4.2 Biofilm 6.11 Literature
4.3 General diagrams for process water 11 Reacting to positive microbiological findings
4.4 Basic pretreatment of incoming water 7 Material transportation and storage 11.1 Basic requirements, official microbiological
4.5 Microbiological treatment of town water 7.1 General conditions thresholds and internal company specifications
4.6 Chemical/physical treatment of incoming water 7.2 Storerooms 11.2 Microbiological analyses and resultant decisions
4.7 Storing treated process water and closed 7.3 Systems and containers 11.3 Basic considerations and validation measures
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4.8 Design for hot process water 7.5 Using packaging and flow materials 11.5 Literature

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 < Content
CO S M ET I C S
SENSITIVE SKIN

Type of Methods Tested Parameters Applications Ref.

Methods
Sociologic - Mail/ telephone queries Self-perception of Epidemiological studies 1-5
queries - Self-assessment questionnaires skin sensitivity
- Visual scales (graded; analogue)
Sensory - Sensory perception to the Subjective sensations: Evaluation of the subjective 6, 7
perception application of chemical lactic acid (stinging); sensory perception
tests substances on the skin capsaicin (burning/stinging),
ethanol (burning and stinging),
menthol (cooling)
- Sensory perception to the Sensory perception 8, 9
application of physical stimuli Thresholds to electrical nerve
stimulation, cold,
and hot stimulation
Objective - Irritation tests: Visible and/or instrumentally Assessment of the objective 10-19
reactivity open application, assessed signs of skin reactivity features of skin sensitivity
tests patch test, strip-patch test,
behind-the-knee epicutaneous Assessed parameters:
test, washing and TEWL; hydration (evaluation
immersion tests of skin electrical properties);
erythema (by colorimetry);
sub-clinical erythema
(by cross-polarized light);
microcirculation/vasodilatation
(by LDI/LDV); surface temperature
(by thermography)
stratum corneum properties
(corneosurfametry, tape-striping)
and components (Sebutape®)
Skin - Biophysical methods TEWL (barrier function); Assessment of cutaneous 15,
functional capacitance/impedance physiological functions 20-23
tests (hydration); skin surface pH;
skin color (colorimetry)
vascular reactivity (LDV/LDI);
sebum lipids (photometric methods)
References:
1. Jourdain R, Lacharriere O, Bastien P , Maibach HI. Ethnic variations in self-perceived sensitive skin: epidemiological survey. Contact Dermatitis 2002;46:162-9.
2. Misery L, Boussetta S, Nocera T, Perez-Cullell N , Taieb C. Sensitive skin in Europe. J Eur Acad Dermatol Venereol 2009;23:376-81.
3. Misery L, Myon E, Martin N, Consoli S, Boussetta S, Nocera T et al. Sensitive skin: psychological effects and seasonal changes. J Eur Acad Dermatol Venereol 2007;21:620-8.
4. Misery L, Myon E, Martin N, Verriere F, Nocera T , Taieb C. [Sensitive skin in France: an epidemiological approach]. Ann Dermatol Venereol 2005;132:425-9.
5. Willis CM, Shaw S, De Lacharriere O, Baverel M, Reiche L, Jourdain R et al. Sensitive skin: an epidemiological study. Br J Dermatol 2001;145:258-63.
6. Frosch PJ , Kligman AM. A method for appraising the stinging capacity of topically applied substances. J Cosmetic Chem 1977;28:197-209.
7. Marriott M, Whittle E , Basketter DA. Facial variations in sensory responses. Contact Dermatitis 2003;49:227-31.
8. Kim SJ, Lim SU, Won YH, An SS, Lee EY, Moon SJ et al. The perception threshold measurement can be a useful tool for evaluation of sensitive skin. Int J Cosmet Sci 2008;30:3337.
9. Stander S, Schneider SW, Weishaupt C, Luger TA , Misery L. Putative neuronal mechanisms of sensitive skin. Exp Dermatol 2009;18:417-23.
10. Berardesca E, Cespa M, Farinelli N, Rabbiosi G , Maibach H. In vivo transcutaneous penetration of nicotinates and sensitive skin. Contact Dermatitis 1991;25:35-8.
11. Farage M , Maibach HI. The vulvar epithelium differs from the skin: implications for cutaneous testing to address topical vulvar exposures. Contact Dermatitis 2004;51:201-9.
12. Dickel H, Kamphowe J, Geier J, Altmeyer P , Kuss O. Strip patch test vs. conventional patch test: investigation of dose-dependent test sensitivities in nickel- and chromium-sensitive sub-
jects. J Eur Acad Dermatol Venereol 2009;23:1018-25.
13. Farage MA. Assessing the skin irritation potential of facial tissues. Cutan Ocul Toxicol 2005;24:125-35.
14. Farage MA. The Behind-the-Knee test: an efficient model for evaluating mechanical and chemical irritation. Skin Res Technol 2006;12:73-82.
15. Fluhr JW, Darlenski R , Berardesca E. Ethnic groups and sensitive skin: two examples of special populations in dermatology. Drug Discovery Today: Disease Mechanisms 2008;5:249-63.
16. Farage MA. Enhancement of visual scoring of skin irritant reactions using cross-polarized light and parallel-polarized light. Contact Dermatitis 2008;58:147-55.
17. Farage MA , Maibach HI. Sensitive skin: closing in on a physiological cause. Contact dermatitis 2010;62:137-49.
18. Perkins MA, Osterhues MA, Farage MA , Robinson MK. A noninvasive method to assess skin irritation and compromised skin conditions using simple tape adsorption of molecular
markers of inflammation. Skin Res Technol 2001;7:227-37.
19. Goffin V, Pierard-Franchimont C , Pierard GE. Sensitive skin and stratum corneum reactivity to household cleaning products. Contact Dermatitis 1996;34:81-5.
20. Wu Y, Wang X, Zhou Y, Tan Y, Chen D, Chen Y et al. Correlation between stinging, TEWL and capacitance. Skin Res Technol 2003;9:90-3.
21. Bornkessel A, Flach M, Arens-Corell M, Elsner P , Fluhr JW. Functional assessment of a washing emulsion for sensitive skin: mild impairment of stratum corneum hydration, pH, barrier
function, lipid content, integrity and cohesion in a controlled washing test. Skin Res Technol 2005;11:53-60.
22. Lonne-Rahm S, Nordlind K, Edstrom DW, Ros AM , Berg M. Laser treatment of rosacea: a pathoetiological study. Arch Dermatol 2004;140:1345-9.
23. Seidenari S, Francomano M , Mantovani L. Baseline biophysical parameters in subjects with sensitive skin. Contact Dermatitis 1998;38:311-5.

Table 2 Methods used in the study of sensitive skin as potential tool in the creation of a successful model of the condition (LDI
– laser-doppler imaging; LDV - laser-doppler velocimetry; TEWL - transepidermal water loss)

20 SOFW-Journal | 136 | 11-2010


was demonstrated in sensitive skin sub-
jects during a repetitive washing test
with an emulsion designed for sensitive
skin (25). However, SC integrity did not
change in the course of the study, indi-
cating that the changes in the surface
lipids had no effect on the structure of
the epidermal barrier.
Electrical current perception threshold
(used to quantify the sensory threshold
to transcutaneous electrical stimulation
of the sensory nerves) was lower in pa-
tients with atopic eczema than in con-
trols (26). This can explain the increased
skin stinging in atopic patients reaching
up to 64% of the affected population
(12). In addition, an increased density of
the cutaneous nerves was shown in
atopic skin (27). These findings, togeth-
er with the elevated number of mast cell
in the lesion of atopic dermatitis can be
involved in the increased skin sensitivity
and inflammatory response in the dis-
ease (12). On the other hand, keratino-
cyte-derived cytokines play an impor-
tant role in the group of contact der-
matoses, i.e. irritant and allergic contact
dermatitis. The secretion of a pre-formed
pool of pro-inflammatory cytokines such
as interleukin 1 and tumor necrosis fac-
tor alpha is crucial in the early phases of
the skin reactivity to chemical irritants
(28).
-Fig. 2 Practical algorithm for the development of sensitive skin model
SOFW-Journal | 136 | 11-2010
< Content
SENSITIVE SKIN
A significant reduction in the LAST score
after laser treatment of rosacea was
demonstrated (22). The molecular inves-
tigations revealed a decrease in sub-
stance P (responsible for vasodilatation)
levels in the papillary dermis after ther-
apy and thus supporting the concept of
increased vessel reactivity in rosacea. In
addition, a reduction of the nerve fiber
network density after treatment was ev-
idenced.
Sensitive skin subjects display abnormal
sensory perception related to neurologi-
cal instability with a major role of c-neu-
ron fibers responsible for nociception
(29). Magnetic resonance imaging study
revealed that subjects with self-per-
ceived sensitive skin have a specific cere-
bral activation during skin irritation test
when compared to controls (30). These
findings suggest that interactions be-
tween processes in the central nervous
systems are involved in the abnormal
sensory response in sensitive skin.
 Multiparametric Approach to
the Development of Sensitive
Skin Model
Not a single physiological or morpho-
logic abnormality can be accepted as the
milestone in sensitive skin syndrome.
CO S M ET I C S
There is no parameter that can fully de-
scribe the complex problem of the sen-
sitive skin. Thus a multiparametric ap-
proach is proposed. Table 2 summarizes
the different methods used in testing
sensitive skin.
Creation of a human model of sensitive
skin is a multi-step process. No single and
robust test method exists. Fig. 2 offers a
practical algorithm for the development
of sensitive skin model.
 Conclusion
Sensitive skin syndrome is a heteroge-
neous condition which is dependent on
multiple factors. Therefore, a necessity
exists for its precise definition, under-
standing the pathophysiological mecha-
nism, and for establishing clear criteria
for its diagnosis. Fulfilling this would en-
able the practical implementation of a
sensitive skin model.
References
(1) Stander S, Schneider SW, Weishaupt C, Luger
TA , Misery L. Putative neuronal mechanisms
of sensitive skin. Exp Dermatol 2009;18:417-
23
(2) Kligman AM. The invisible dermatoses. Arch
Dermatol 1991;127:1375-82
(3) Farage MA, Katsarou A, Maibach HI. Senso-
ry, clinical and physiological factors in sensi-
tive skin: a review. Contact Dermatitis
2006;55:1-14
(4) Issachar N, Gall Y, Borell MT , Poelman MC.
pH measurements during lactic acid stinging
test in normal and sensitive skin. Contact Der-
matitis 1997;36:152-5
(5) Kligman AM, Sadiq I, Zhen Y, Crosby M. Ex-
perimental studies on the nature of sensitive
skin. Skin Res Technol 2006;12:217-22
(6) Mills OH, Jr., Berger RS. Defining the suscep-
tibility of acne-prone and sensitive skin pop-
ulations to extrinsic factors. Dermatol Clin
1991;9:93-8
(7) Draelos ZD. Is this product designed for sen-
sitive skin? Cosmetic Dermatology 2002;15:71-
8
(8) Willis CM, Shaw S, De Lacharriere O, Baverel
M, Reiche L, Jourdain R et al. Sensitive skin:
an epidemiological study. Br J Dermatol
2001;145:258-63
21

CO S M ET I C S
SENSITIVE SKIN
(9) Jourdain R, Lacharriere O, Bastien P, Maibach
HI. Ethnic variations in self-perceived sensi-
tive skin: epidemiological survey. Contact
Dermatitis 2002;46:162-9
(10) Misery L, Myon E, Martin N, Consoli S, Bous-
setta S, Nocera T et al. Sensitive skin: psycho-
logical effects and seasonal changes. J Eur
Acad Dermatol Venereol 2007;21:620-8
(11) Misery L, Myon E, Martin N, Verriere F, Nocera
T , Taieb C. [Sensitive skin in France: an epi-
demiological approach]. Ann Dermatol Venere-
ol 2005;132:425-9
(12) Lonne-Rahm S, Berg M, Marin P, Nordlind K.
Atopic dermatitis, stinging, and effects of
chronic stress: a pathocausal study. J Am Acad
Dermatol 2004;51:899-905
(13) Berg M, Lonne-Rahm SB , Fischer T. Patients
with visual display unit-related facial symp-
toms are stingers. Acta Derm Venereol 1998;
78:44-5
(14) Andreassi L. Bioengineering in dermatology:
general aspects and perspectives. Clin Der-
matol 1995;13:289-92
(15) Distante F, Rigano L, D'Agostino R, Bonfigli A,
Berardesca E. Intra- and Inter- Individual Dif-
ferences in Sensitive Skin. Cosmetics & Toi-
letries magazine 2002;117:39-46
(16) Wu Y, Wang X, Zhou Y, Tan Y, Chen D, Chen Y
et al. Correlation between stinging, TEWL and
capacitance. Skin Res Technol 2003;9:90-3
(17) Seidenari S, Francomano M, Mantovani L.
Baseline biophysical parameters in subjects
with sensitive skin. Contact Dermatitis 1998;
38:311-5
(18) An S, Lee E, Kim S, Nam G, Lee H, Moon S et
al. Comparison and correlation between
stinging responses to lactic acid and bioengi-
neering parameters. Contact Dermatitis
2007;57:158-62
(19) Berardesca E, Cespa M, Farinelli N, Rabbiosi
G, Maibach H. In vivo transcutaneous pene-
tration of nicotinates and sensitive skin. Con-
tact Dermatitis 1991;25:35-8
22
< Content
(20) Sparavigna A, Pietro A, Setaro M. Sensitive (29) Kim SJ, Lim SU, Won YH, An SS, Lee EY, Moon
skin: correlation with skin surface microrelief SJ et al. The perception threshold measure-
appearance. Skin Res Technol 2006;12:7-10 ment can be a useful tool for evaluation of
sensitive skin. Int J Cosmet Sci 2008;30:333-
(21) Issachar N, Gall Y, Borrel MT, Poelman MC. 7
Correlation between percutaneous penetra-
tion of methyl nicotinate and sensitive skin, (30) Querleux B, Dauchot K, Jourdain R, Bastien P,
using laser Doppler imaging. Contact Der- Bittoun J, Anton JL et al. Neural basis of sen-
matitis 1998;39:182-6 sitive skin: an fMRI study. Skin Res Technol
2008;14:454-61
(22) Lonne-Rahm S, Nordlind K, Edstrom DW, Ros
AM , Berg M. Laser treatment of rosacea: a (31) Draelos ZD. Treating the patient with multi-
pathoetiological study. Arch Dermatol 2004; ple cosmetic product allergies. A problem-
140:1345-9 oriented approach to sensitive skin. Postgrad
Med 2000;107:70-2, 5-7
(23) Fluhr JW, Kao J, Jain M, Ahn SK, Feingold KR,
Elias PM. Generation of free fatty acids from (32) Wolff K, Goldsmith LA, Katz SI, Gilchrest BA,
phospholipids regulates stratum corneum Paller A, Leffell DJ. Fitzpatrick's Dermatology
acidification and integrity. J Invest Dermatol in General Medicine, 7th edition: McGraw-Hill
2001;117:44-51 Book Co; 2008
(24) Schmid-Wendtner MH, Korting HC. The pH of
the skin surface and its impact on the barri-
er function. Skin Pharmacol Physiol 2006;
19:296-302 Authors addresses:
* Joachim W. Fluhr, MD
(25) Bornkessel A, Flach M, Arens-Corell M, Elsner Department of Dermatology
P, Fluhr JW. Functional assessment of a wash- Charité Université Clinic
ing emulsion for sensitive skin: mild impair-
ment of stratum corneum hydration, pH, bar-
Charitéplatz 1
rier function, lipid content, integrity and co- 10117 Berlin
hesion in a controlled washing test. Skin Res Germany
Technol 2005;11:53-60
** Razvigor Darlenski
(26) Kobayashi H, Kikuchi K, Tsubono Y , Tagami
H. Measurement of electrical current percep-
Department of Dermatology and
tion threshold of sensory nerves for pruritus Venerology
in atopic dermatitis patients and normal in- Medical Faculty
dividuals with various degrees of mild dam- University of Medicine
age to the stratum corneum. Dermatology Sofia
2003;206:204-11
Bulgaria
(27) Urashima R, Mihara M. Cutaneous nerves in
atopic dermatitis. A histological, immunohis- Correspondence address:
tochemical and electron microscopic study. Joachim W. Fluhr, MD
Virchows Arch 1998;432:363-70 Email: joachim.fluhr@gmx.net
(28) Fluhr JW, Darlenski R, Angelova-Fischer I,
Tsankov N, Basketter D. Skin Irritation and 
Sensitization: Mechanisms and New Approach-
es for Risk Assessment. Part 1: Skin irritation.
Skin Pharmacol Physiol 2008;21(3):124-35
SOFW-Journal | 136 | 11-2010
 < Content

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CO S M ET I C S
T ET R A P E PT I D E
M. Mateu*, E. Cañadas*, J. Cebrián*, C. Carreño**, N. Almiñana*, R. Delgado*
A New Tetrapeptide for Elasticity and
Tightness Enhancement
 Introduction
A variety of environmental, hormonal,
and genetic factors results in skin elas-
ticity loss. Mature skin becomes less elas-
tic and less able to resist any deforma-
tion leading to many of the visible man-
ifestations of aging such as wrinkles, un-
firmness or sagginess (1).
Skin quality deteriorates with age due to
the synergistic effects of chronological
aging, photoaging, environmental fac-
tors, and hormonal deficiency. Hormon-
al aging of skin due to oestrogen loss at
menopause is thought to include atro-
phy, elasticity loss and decreased seba-
ceous secretions, and collagen and wa-
ter content (2).
Intrinsically aged skin shows character-
istic fine wrinkling and appears smooth
(3). Especially from the age of 40 years,
synthesis and turnover of new compo-
nents by fibroblasts slow and enzyme ac-
tion on fibres increases implying skin
elasticity loss and a less supple and more
hardened collagen. On the other hand,
solar radiation is the main responsible
for extrinsic aging, but some other caus-
es, such as air pollution, are also very im-
portant (4). Severe photoaging presents
increased proteolytic activation and
shows abnormal extracellular matrix
(ECM) turnover. The consequence is an
acceleration of collagen and elastic fi-
bres degradation in the dermis resulting
in loss of skin's ability to resist stretch-
ing (5). Typically, sun-exposed skin ap-
pears papular, coarse, roughened, and
deeply wrinkled with marked loss of
elasticity and recoil (6).
Elastic fibres are responsible for the nor-
mal resilience of the skin. When elastic
24
< Content
fibres suffer elongation, their immediate tissues withstand stretching (7). One of
tendency is to return to their initial po- the most important is type I collagen,
sition with an elastic behaviour (Fig. 1). which is the most abundant collagen in
This elasticity decreases with time for the human body, representing the 80-
different reasons such as natural aging 85% of the dermal collagen. Type I col-
or other several factors that accelerate lagen fibrils have a great tensile strength
or modify the natural process (4). and elastic resistance (1, 6).
Skin elasticity is a mechanical property Elastic fibres are insoluble structural el-
which is influenced by elastin, a protein ements of connective tissue that have a
in the skin which, together with collagen central core of amorphous, hydrophobic
and glycosaminoglycans, make up the cross-linked elastin surrounded by fibril-
connective tissue. Protein fibres are lar structures with a regular diameter of
arranged to form a network submerged 10-12 nm (Fig. 2). Elastin is a well-char-
in a gel matrix of water and glycosamino- acterised connective tissue protein and
glycans. Fibroblasts are included in this
structure and are responsible for synthe-
sising the other components. The protein
network gives the tissue its physical
properties, such as rigidity and elastici-
ty, elastin being fundamental in this lat-
ter parameter (4).
The connective tissue of the skin is com-
posed mostly of collagen which is the
most abundant protein in the skin. Col-
lagen makes up 70-80% of the dry weight
of the skin and gives the dermis its me-
chanical and structural integrity (6). The -Fig. 2 Elastin core surrounded by fib-
various collagens and the structures they
form all serve the same purpose, to help rillar structures
-
Fig. 1 Relaxed elastic fibres (left) and stretched elastic fibres (right)
SOFW-Journal | 136 | 11-2010
< Content

CO S M ET I C S
T ET R A P E PT I D E
is the major component of the elastic fi-
bres. Although, elastin is found in less
amount than collagen in the dermis, it is
crucial for skin elasticity. During early
embryonic development, most of the
elastic fibres consist of microfibrils, which
form a microfibrillar skeleton upon in
which elastin is deposited. In mature,
fully developed elastic tissue, well over
90% of the total content consists of
elastin (3, 6).
Natural aged skin shows general atrophy
of the ECM with reduced elastin and dis-
integration of elastic fibres (3). There is
also a reduction in the amount of peri-
pheral microfibrils. The fibre surface be-
comes irregular and granular, microfib-
rils become thicker, and there is a de-
crease in the amount of glycosaminogly-
cans and fibroblasts (4). A major feature
of aged skin is also reduced collagen syn-
thesis and increased degradation, result-
ing in connective tissue damage, and loss
of the skin three-dimensional integrity
(8).
Chronological aging process and envi-
ronmental insults contribute to genera-
tion of ROS (Reactive Oxygen Species)
that stimulate the inflammatory process
in the skin activating the transcription
factors that regulate the proteolytic
degradation of the ECM. In response to
UV-induced production of pro-inflam-
matory cytokines, phagocytic cells such
as neutrophils and monocytes infiltrate
into skin from capillaries. In addition to
keratinocytes, the phagocytic cells them-
selves secrete cytokines that further en-
hance recruitment of inflammatory cells.
Furthermore, neutrophils release elas-
tases and other proteases that can cause
further inflammation, and activation of
matrix metalloproteases (MMP) which are
known for degrading collagen fibres (5).
Damage to connective tissues is a major
complication of the inflammatory re-
sponse. Elastic fibres are degraded by sev-
eral types of enzymes, such as neutrophil
elastase released during neutrophil infil-
tration of the epidermis, MMP-12 de-
rived from macrophages, and skin fi-
broblast elastase produced by fibroblasts
(5, 9).
Ultraviolet radiation induces both neu-
trophil elastase and skin fibroblast elas-
tase. Neutrophil elastase is able to rapid-
ly degrading intact microfibrils and its
26
< Content
inhibition has demonstrated to prevent
UV-induced wrinkle formation in skin.
The secretion and activation of skin fi-
broblast elastase is thought to be re-
sponsible for the degeneration of the
three dimensional structure of elastic fi-
bres during the formation of wrinkles (3,
5, 9).
Inflammation, UV irradiation and normal
process of aging can activate MMPs
leading to increased matrix degradation.
MMPs are a group of zinc-dependent en-
dopeptidases capable of degrading ECM
components and are involved in the
turnover and remodelling of the dermis
(5). Human macrophage metalloelastase
(HME, MMP-12) is the most active MMP
against elastin on a molar basis and has
broad substrate specificity, being able to
degrade also type IV collagen which is
the most abundant structural compo-
nent of basement membranes (10). On
the other hand, MMP-1 initiates cleav-
age of fibrillar collagen types I and III in
the dermis, which is then further de-
graded by MMP-2 and -9 (5). An increase
of elastases activity and a slow elastoge-
nesis end up in elasticity loss which is re-
flected in a sagging, unfirmed and wrin-
kled skin.
A new tetrapeptide identified by a com-
binatorial chemistry approach from a li-
brary of 331776 peptides, showed to
posses skin elasticity and tightness en-
hancement properties. The combinator-
ial peptide library was screened by mon-
itoring fluorescence of quenched elastin
released when digested by elastase to
evaluate the inhibitory potency of the
peptides on the activity of elastase en-
zyme.
Relistase™ (INCI name Acetylarginyltryp-
tophyl Diphenylglycine) proved in vitro
to inhibit the excess of elastase activity,
helping to improve skin elasticity lost
due to aging. By reducing the excess of
elastase activity, it helps to protect elastin
and other ECM components which are
susceptible to be degraded by these en-
zymes. The tetrapeptide also presented
collagen boosting properties in vitro which
favour connective tissue improvement
and helps to restore skin three dimen-
sional integrity by enhancing tensile
strength and elastic resistance. More-
over, the tetrapeptide showed in vivo to
improve skin elasticity and tightness.
 Materials and Methods
Elastin Protection from Pig Pancreatic
Elastase
The fluorescence released by quenched
elastin when digested by pig pancreatic
elastase was monitored in order to study
Acetylarginyltryptophyl Diphenylglycine
dose-response inhibition of the porcine
elastase.
Samples at 1, 10 and 50 µM were prein-
cubated with the protease reconstituted
in Reaction Buffer (0.4 units/mL of pig
pancreatic elastase) for 1 hour at room
temperature. After pre-incubation, 25 µL
of the substrate (DQ Elastin) were added
and the samples were incubated in dark-
ness for 2 hours at room temperature.
Fluorescence released by the digestion of
labelled elastin was measured in an au-
tomated multiplate fluorescence reader
set for excitation at 485 nm and detec-
tion at 530 nm. The results obtained were
corrected from the basal fluorescence
released with neither elastase nor test
items and normalised regarding the re-
lease of fluorescence of a control exper-
iment without test items (negative con-
trol).
Inhibition of Human Neutrophil Elastase
The fluorescence released by the fluoro-
genic elastase substrate V (MeOSuc-Ala-
Ala-Pro-Val-aminomethylcoumarin) when
digested by human neutrophil elastase
was monitored in order to study Acetyl-
arginyltryptophyl Diphenylglycine dose-
response inhibition of the human neu-
trophil elastase.
Samples at 50, 100 and 500 µM were
preincubated with 0.2 µg/mL of human
neutrophil elastase in Reaction Buffer
for 1 hour at room temperature. After-
wards, the fluorogenic substrate was
added to wells and samples were incu-
bated in darkness for 2 hours at room
temperature. The hydrolysis of the fluo-
rogenic elastase substrate V was moni-
tored fluorometrically with a 370 nm ex-
citation filter and a 460 nm emission fil-
ter in an automated multiplate fluores-
cence reader. The results obtained were
corrected from the basal fluorescence re-
leased with neither elastase nor test items
and normalised regarding the release of
fluorescence of a control experiment
without test items (negative control).
SOFW-Journal | 136 | 11-2010
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16–17 DECEMBER 2010

MUMBAI, INDIA
HOME AND PERSONAL CARE INGREDIENTS CONGRESS
GRAND HYATT HOTEL, MUMBAI, SANTACRUZ EAST

The HPCI Congress, freaturing a trade exhibition

with a scientific conference, is set to become the
most comprehensive show for home and personal
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The HPCI Exhibition offers its visitors the excep-
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Indian Society of Cosmetics Chemists SOFW – Verlag für chemische Industrie, H. Ziolkowsky GmbH · Beethovenstrasse 16
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CO S M ET I C S
T ET R A P E PT I D E

Evaluation of Type I Collagen Induction manner, showing an elastase inhibition Cutaneous Elasticity and Tightness
on Human Dermal Fibroblasts of 86% at 500 µM (Fig.4). Evaluation
Collagen induction by Acetylarginyltryp- The cream containing the active ingredi-
tophyl Diphenylglycine was evaluated by Evaluation of Type I Collagen Induction ent showed a highly significant improve-
an Enzyme-linked Immunosorbent Assay on Human Dermal Fibroblasts ment on the overall elasticity of 11.7%
(ELISA). Type I collagen from the culture The peptide proved to increase by 99% and 14% after 4 and 8 weeks respectively
medium was attached to the bottom of type I collagen synthesis induction on (Fig.6). No statistically significant varia-
a plate well. It was detected with an an- human dermal fibroblasts cell cultures at tion was detected in the values of the
ti-collagen type I antibody. This antibody 21 µM (Fig.5). area treated with placebo cream.
was recognised by a labelled secondary
antibody. The assay was then quantified
by measuring the amount of labelled an-
tibody bound to the matrix, by using a
colorimetric substrate. Peroxidase labelled
to the secondary antibody converts the
colourless substrate (OPD) to a coloured
product. This colour was measured and it
is proportional to the quantity of type I
collagen present in the sample.

Cutaneous Elasticity and Tightness

Evaluation
A panel of volunteers composed of 20
women (mean age 49 years) applied a
cream containing 0.0004% Acetylarginyl-
tryptophyl Diphenylglycine or a placebo
cream on their thighs twice a day for 8
weeks.
Skin elasticity was determined by a Cuto-
meter SEM 575, Courage & Khazaka which
measures the vertical deformation of the -Fig. 3 Inhibition percentage of elastase activity by Acetylarginyltryptophyl
skin, when it is sucked into the opening
Diphenylglycine at 1, 10 and 50 µM. Data are shown as MEAN
of a measuring probe. Mean values and
standard deviations were calculated for
T0, T4 and T8 instrumental values. In-
strumental data and the variations were
statistically compared by means of paired
samples t-test. The differences were con-
sidered significant when the probability
p was ≤ 0.05.

 Results and Discussion

Elastin Protection from Pig Pancreatic

Elastase
Under the experimental conditions, the
tetrapeptide proved to be able to protect
elastin by inhibiting pig pancreatic elas-
tase activity in a dose-response manner,
showing an inhibition of 80.2% at 50 µM
(Fig. 3).

Inhibition of Human Neutrophil Elastase

Acetylarginyltryptophyl Diphenylglycine -Fig. 4 Inhibition percentage of human neutrophil elastase activity by the peptide
demonstrated to be able to inhibit hu- at 50, 100 and 500 µM. Data are shown as MEAN
man neutrophil elastase in a dose-response

28 SOFW-Journal | 136 | 11-2010

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CO S M ET I C S
T ET R A P E PT I D E

The area treated with the cream con-

taining the tetrapeptide showed a high-
ly significant decrease in maximal defor-
mation values of -5.5% and -15.6% af-
ter 4 and 8 weeks of treatment (Fig.7.).
However, no statistically significant vari-
ation was detected in the values of the
area treated with placebo cream.

 Conclusions

In mature skins, elastin synthesis de-

creases and its degradation enhances re-
sulting in elasticity loss. Stimulation of
dermal fibroblasts not only induces
elastin production but upregulate elas-
totic enzymes which may rapidly degra-
date newly produced elastin and existing
elastic fibres. Hence, there is a need to
protect existing and new elastic fibres -Fig. 5 Increase of type I collagen synthesis induced by 21 µM Acetylarginyltrypto-
from premature enzymatic proteolysis in phyl Diphenylglycine respect to non-treated cells
order to get a firm, elastic and wrinkless
skin (11).

3 rd
Edition
COSMETAGORA
The meetings of Formulation
January 19th & 20th, 2011
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You are a project leader, you work in a laboratory, you

carry out tests, you are student… This show is for you!

You can download invitations on:

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SOFW-Journal | 136 | 11-2010 29

 < Content
CO S M ET I C S
T ET R A P E PT I D E

At the light of the results, it can be con-

cluded that Acetylarginyltryptophyl Di-
phenylglycine is a good cosmetic active
ingredient for body skin elasticity and
tightness enhancing cosmetic formula-
tions especially targeted at mature skins.

References

(1) Pons Gimier L, Parra Juez JL. Ciencia Cos-

mética. Bases fisiológicas y criterios prácticos.
Consejo General de Colegios Oficiales de Far-
macéuticos. 1995

(2) Brincat MP. Oestrogens and the skin. J Cos-

met Dermatol. 3(1): 41-49, 2004

(3) Watson RE, Griffiths CE, Craven NM, Shuttle-

worth CA, Kielty CM. Fibrillin-rich microfib-
rils are reduced in photoaged skin. Distribu-
-Fig. 6 Overall elasticity of the skin after applying a placebo or a cream containing tion at the dermal-epidermal junction. J In-
vest Dermatol. 112(5): 782-787, 1999
0.0004% Acetylarginyltryptophyl Diphenylglycine
(4) Benaiges A, Marcet P, Armengol R, Betes C,
Gironés E. Study of the refirming effect of a
Acetylarginyltryptophyl Diphenylglycine three-dimensional integrity and con- plant complex. Int J Cosmet Sci. 20(4): 223-
233, 1998
proved to protect existing elastin by in- nective tissue improvement thanks to its
hibiting human neutrophil elastase ac- excellent collagen booster properties
(5) Pillai S, Oresajo C, Hayward J. Ultraviolet ra-
tivity, in vitro, helping to avoid elastici- which showed in vitro. In addition, the
diation and skin aging: roles of reactive oxy-
ty loss due to elastin and other ECM com- tetrapeptide demonstrated in vivo to sig- gen species, inflammation and protease acti-
ponents degradation by elastase. More- nificantly enhance skin elasticity and vation, and strategies for prevention of in-
over, the peptide favours to restore skin tightness of the body. flammation-induced matrix degradation – a
review. Int J Cosmet Sci. 27(1): 17-34, 2005

(6) Oikarinen A. Aging of the skin connective tis-

sue: how to measure the biochemical and me-
chanical properties of aging dermis. Review.
Photodermatol Photoimmunol Photomed.
10(2): 47-52, 1994

(7) Culav EM et al. Connective tissues: matrix

composition and its relevance to physical
therapy. Phys Ther. 79: 308-319, 1999

(8) Bernstein EF et al. Long-term sun exposure

alters the collagen of the papillary dermis. J
Am Acad Dermatol. 34: 209-218, 1996

(9) Tsukahara K, Takema Y, Moriwaki S, Tsuji N,

Suzuki Y, Fujimura T, Imokawa G. Selective in-
hibition of skin fibroblast elastase elicits a
concentration-dependent prevention of ul-
traviolet B-induced wrinkle formation. J In-
vest Dermatol. 117(3): 671-677, 2001. Erra-
tum in: J Invest Dermatol . 118(4): 742, 2002

(10) Saarialho-Kere U, Kerkelä E, Jeskanen L,

-Fig. 7 Maximal deformation of the skin after applying a placebo or a cream con- Hasan T, Pierce R, Starcher B, Raudasoja R,
taining 0.0004% Acetylarginyltryptophyl Diphenylglycine Ranki A, Oikarinen A, Vaalamo M. Accumula-

30 SOFW-Journal | 136 | 11-2010

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CO S M ET I C S
T ET R A P E PT I D E

tion of matrilysin (MMP-7) and macrophage

metalloelastase (MMP-12) in actinic damage.
J Invest Dermatol. 113(4): 664-672, 1999
SOFW
(11) Jimenez F, Mitts TF, Liu K, Wang Y, Hinek A.
Ellagic and tannic acids protect newly syn- Media Information 2011
thesized elastic fibers from premature enzy-
matic degradation in dermal fibroblast cul-
tures. J Invest Dermatol. 126(6): 1272-1280,
2006

Authors’ addresses:
* Miriam Mateu, Elena Cañadas
Juan Cebrián, Núria Almiñana
Raquel Delgado
Lipotec S.A.
Pol. Ind. Cami Ral. Isaac Peral, 17
08850 Gavá
Spain
Email: mmateu@lipotec.com

** Cristina Carreño
DiverDrugs S.L.
Pol. Ind. Cami Ral. Isaac Peral, 17
08850 Gavá
Spain
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SOFW-Journal | 136 | 11-2010 31


CO S M ET I C S
HAIR CARE
Y. Kamath*, V. Gode**, N. Bhalla**, B. Kalghatgi**, S. Mhaskar**
Effect of Grooming Practices on
Physical Characteristics of Hair
 Introduction
Various research papers have been pub-
lished on the physical and biochemical
characteristics of hair. Although racial
he grooming habits differ sig-
T
nificantly amongst various eth-
nic groups. Oiling the hair as
part of the grooming habit is
prevalent mostly in the South East
Asian countries and not in the
western world or amongst African
populations. Vegetable oils have
been used for hair care for several
years. These oils are believed to
improve appearance and the ten-
sile properties of hair.
Objective: To determine whether
differences in grooming practices in
different ethnic populations have
an impact on the physical charac-
teristics of hair.
Methods: Hair samples each ob-
tained from fifteen European,
African and Indian volunteers were
collected by cyberDerm (Broomall,
PA). The impact of grooming habit
on the hair of these populations
32
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differences in hair characteristics have
been the subject of much interest, there
are relatively fewer research papers pub-
lished on the ethnic differences in physi-
cal properties of hair (1-7).
Abstract
was determined by measuring vari-
ous physical properties. The area of
cross section, and the tensile prop-
erties were measured using Dias-
tron™. Hairs were conditioned at
65% RH and 21 °C overnight before
the measurements. Hair morpholo-
gy was assessed using a scanning
electron microscope (SEM). Protein
loss analysis was conducted by de-
termining the extent of protein elu-
sion from the hair samples using
the Lowry method. Porosity of hair
was measured using Valko and Bar-
nett method.
Results: Cross sectional area of the
Indian hair samples was signifi-
cantly higher than the African and
Caucasian hair. The elastic modulus
values of Indian hair were also
higher suggesting that hair break-
age during combing is lower in In-
dian hair. Indian hair samples were
Human hair is categorized into three
major groups based on ethnic origin:
Asian, African and Caucasian. Various re-
searchers have demonstrated that there
are no major biochemical differences of
significantly less porous as com-
pared to the Caucasian and African
hair. Caucasian as well as African
hair also had lower luster values
than Indian hair.
Conclusion: Oiled Indian hair stud-
ied during this investigation had
significant higher values for cross
sectional area and for other physi-
cal parameters such as tensile
strength and luster. Oiled Indian
hair also demonstrated lower
porosity and lower protein loss
than Caucasian and African hair.
The data obtained from unoiled
hair samples was comparable to
those of the African and Caucasian
counterparts suggesting that
though there were genetic differ-
ences in hair types these differ-
ences became more significant
when grooming was included as a
factor affecting the quality of hair.
SOFW-Journal | 136 | 11-2010
 < Content

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CO S M ET I C S
HARE CARE
hair between the various ethnic groups
(6, 7). However, a few studies have demon-
strated that there is significant differ-
ence in the physical properties of hair
from different ethnic populations (1).
Multiple factors such as current and his-
torical cultural practices of grooming
hair is thought to have an important role
in this difference in physical properties,
in addition to the genetic and geo-
graphic variations. For example, heat re-
structuring was a common hair care
practice used by Africans in the past. Hot
combing was the first method by which
individuals were able to restructure or
straighten the hair unit by temporari-
ly manipulating the hydrogen disulfide
bonds. Hot combing has become less
popular with the introduction of chemi-
cal relaxers for hair straightening. These
relaxers cleave and rearrange the disul-
phide bonds leading to potential prob-
lems such as local contact irritation,
local chemical burns, loss of tensile
strength and increased fragility of hair
shaft (8).
Oiling the hair shaft is an integral com-
ponent of traditional hair care practices
in many Asian communities, especially
southern region of India. Prolonged use
of Coconut oil for hair is known to pro-
tect and improve the quality of the hair
(9). This investigation was designed to
understand differences in hair from dif-
ferent ethnicities and to determine if the
grooming habits in particular oiling the
hair shaft had an influence on the quali-
ty of hair.
 Materials and Methods
Study design
Hair samples were obtained from female
volunteers in the age group of 25-35
years of three different ethnic cate-
gories – African, Caucasian and Asian by
cyberDerm (Broomall, PA). A total of 15
samples were selected from each Cau-
casian and African Hair whereas total of
30 samples were selected from Indian
Hair, 15 from oil users and 15 from non
users. The hair samples included a shaft
length of 7 cm leaving behind 1cm of
hair from the scalp. All the samples re-
ceived a pretreatment of washing with a
clarifying shampoo, rinsing and air dry-
34
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ing before being tested. The African vol-
unteers consisted of people who regu-
larly used straighteners and neutralizers
on their hair. The Caucasian population
consisted of volunteers who used sham-
poos and conditioners for grooming
their hair. Asian hair samples used in this
study were of Indian origin which in-
cluded prolonged Coconut oil users and
non users. In order to ascertain whether
the differences in hair were truly due to
the oiling practice of grooming, Indian
oiled hair was also compared to unoiled
hair. Certain key properties like cross-
sectional area, tensile strength and pro-
tein loss were measured.
Measurements
Scanning electron microscope (SEM)
measurement: Thirty hairs from each
volunteer were taken at random and pre-
pared as knots. These were fixed on a slit-
ted stub and sputter coated with gold.
These were analyzed in a VP-SEM (S-
3000N, Hitachi).
Hair porosity determination: The hair
porosity was determined using a modi-
fied Valko and Barnet method. (10)All
samples were equilibrated at 65% rela-
tive humidity (RH) and 21 °C for 24 hours.
To begin the porosity measurement each
sample was weighed at 65% RH using a
Shimadzu microbalance (AW-220, Shi-
madzu) with a least count of 1 milligram.
The samples were immersed in 100 mL of
distilled water for 30 minutes, removed
with stainless steel forceps and placed on
a filter paper. The hair was dabbed and
kept for 10 minutes in same condition.
The samples were weighed again (final
weight) on the microbalance. Repro-
ducible results were obtained by this
method for each of the sample. The
porosity was calculated using formula:
%Porosity = {[Wa-(W-0.162W)]/Wa}*100
where
W = Weight of hair sample at 65%RH
Wa = Final weight of hair sample
W-0.1625W is described by Menkart et al. (11)
Area of cross section and Tensile strength:
Hair samples were conditioned overnight
at 65% RH and temperature of 21 °C be-
fore the measurements were done. The
cross section of each fibre was measured
using Laser micrometry technique. For
tensile testing, 3 cm long fibres were
mounted on Diastron crimps and were
stretched to break on the automatic Di-
astron Miniature Tensile tester. The en-
tire data for tensile strength was evalu-
ated by using MTTWIN software. From
the load-extension curves, relevant me-
chanical properties were calculated. A
load of 10 N was applied to the fibres and
the operating speed was 10 mm/min. Re-
sults were statistically evaluated using
T- test, Tukey-Kramer HSD and Dunnett’s
method.
Protein loss analysis: Protein loss, an in-
dicator of hair damage was studied to
check the integrity of different samples
and to correlate with strength of hair.
This was analyzed by the method de-
scribed by Sandhu and Robbins (12). Two
grams of hair from each volunteer was
used for analysis. The hair was washed
with 15% sodium laureth sulphate (SLES)
solution and rinsed under running water.
The wet hair was thoroughly combed
with a fine toothed nylon comb (20-22
teeth per inch), for 50 times along the
entire length of the tress. After every 5
strokes the comb was dipped in 50 mL of
water contained in a beaker to dislodge
the debris. After every ten strokes the
entire tress was dipped in the beaker of
water to collect the damaged and dis-
lodged cuticle cells. At the end of 50
strokes the entire tress was dipped in the
beaker and sonicated so that the entire
dislodged protein was transferred into
the water. The water suspension was test-
ed for protein content using the Lowry
SOFW-Journal | 136 | 11-2010
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CO S M ET I C S
HAIR CARE

method. This method involves the for-
mation of a copper protein complex
in alkaline solution that reacts with Fo-
lin Ciocalteau phenol reagent to yield
an intense blue colored solution. This was
spectrophotometrically analyzed at
750 nm.
Luster measurement: Luster values for
each hair fiber were measured using a
modified Brice-Phoenix Universal Light-
Scattering Photometer with He-Ne laser
as the light source. The luster value (L)
was calculated using the formula:
2.8S
L=
(S+D)
where
S = Deconvoluted specular peak area
D = Area of the diffuse component
(S+D) = Total area under the reflection curve
W1/2 = Normalized width at half maximum
47% followed by Caucasian with 41%
and Indian hair with 35% respectively.
High water uptake of African and Cau-
casian hair might be a result of damage
caused to the hair. Lower porosity values
for Indian hair suggest that Indian hair
is less damaged as compared to Cau-
casian and African hair samples. As re-
ported by Ruetsch et al. (14) vegetable
oils penetrate into various levels of cu-
ticular scales. These oils may form a bar-
rier below the cuticle and prevent the
entry of water into the hair shaft. Simi-
W1/2
lar effect of oil on the Water retention
Index (WRI) in oiled and unoiled hair has
also been reported (15). Hence it is like-
ly that in Indian hair the oil applied to
the hair remains below the cuticle even
post wash and reduces the porosity val-
ues.
A good correlation has been observed
between the data from the experiments
conducted on porosity and the SEM im-
ages. Both these results seem to indicate
that the African hair is more damaged as
compared to the Caucasian and Indian
hair.
Cross-sectional area
This attribute is of major significance in
the behavior of hair assemblies. It con-
veys volume, liveliness and resilience. The
cross-sectional areas of hair samples
from the three different ethnic groups
are given in Table 1 and Fig. 3. Judging
by the confidence range (95% confi-
dence level) and confirmed by the sta-

All the parameters were extracted from

the light reflection profile using the Peak
Fit™ software (13).

 Results and Discussion

The results obtained from the experi-

ments above were statistically analyzed
to assess the significance values for the
differences in samples. These results are
summarized below. -Fig. 1 SEM images of knots. (a) African hair (b) Caucasian hair (c) Indian hair

Hair morphology
Fig. 1 shows SEM images of knots in
African, Caucasian and Indian hair. SEM
images generally reveal the status of cu-
ticle scales or defects in the cortex de-
pending on the degree of damage. SEM
examination of the knots revealed that
African hairs were structurally more
damaged, with uplifted cuticle scales
and frayed hair shafts as compared to
Caucasian hair. There was evidence of
structural damage to the hair shaft and
also cuticles were well aligned in the In-
dian hair samples.

Hair porosity
It is apparent from Fig. 2 that African -Fig. 2 Porosity of hairs in various ethnic groups
hairs have the highest porosity of about

SOFW-Journal | 136 | 11-2010 35

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CO S M ET I C S
HARE CARE

Cross-sectional area (m2)

Hair Type Mean ± SE

Indian 5.361e-9 ± 0.1540e-9 a

Caucasian 3.9056e-9 ± 0.08485e-9 b

African 4.3061e-9 ± 0.2007e-9 b

Values in different letters in superscript are significantly different (p<0.05)

Table 1 Cross-sectional area of hair in various ethnic groups -Fig. 3 Cross-sectional area of hair in various ethnic groups

Cross-sectional area (m2)

Hair Type Mean ± SE

Indian (Oil user) 5.3461e-9 ± 0.1540e-9 a

Indian (Non oil user) 4.8301e-9 ± 0.1175e-9 b

Values in different letters in superscript are significantly different (p<0.05)

Table 2 Cross-sectional area of Indian hair -Fig. 4 Cross-sectional area of oiled and unoiled Indian hair

tistical tool, the diameter of Indian hair
samples were significantly more than
Caucasian and African hair samples.
Table 2 and Fig. 4 denotes the difference
in the area of cross section between In-
dian oiled and unoiled hair samples. As
seen from the data oiled Indian hair has
a higher area of cross section as com-
pared to the non oiled hair.
Tensile strength
Data given in Table 3 and Fig. 5 show that
the mechanical properties of hair from
Caucasian samples are better compared
to those of Indian and African hair sam-
ples.
Table 4 provides comparison between
Indian oiled and unoiled hair. Indian
oiled hair showed a lower elastic modu-
lus than unoiled hair. Penetration of oil
into the cuticular scales of hair prevents
water from entering into the fibre. Oil
also causes plasticization of hair fibres.
Hence modulus of oil users is less than
non users. The same phenomenon can be
attributed to break extension where hair
treated with oil break at higher exten-
sion than hair not treated with oil.

Elastic Work for 15% Total work to

Modulus (GPa) extension (MJ/m2) break (M/Jm2)
Mean ± SE Mean ± SE Mean ± SE
Indian 2.73 ± 0.02 a 0.402 ± 0.002a 1.98 ± 0.02a
Caucasian 2.86 ± 0.01 b 0.412 ± 0.001b 2.50 ± 0.02b
African 2.48 ± 0.05 b 0.350 ± 0.008 c 1.50 ± 0.06 c

Values in different letters in superscript are significantly different (p<0.05)

Table 3 Tensile strength of hair in various ethnic groups

-Fig. 5 Total work to break hairs of various ethnic groups

36 SOFW-Journal | 136 | 11-2010

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CO S M ET I C S
HARE CARE

Protein Loss
Cuticle chipping that result from abra- Elastic Work for 15% Total work to Break
sion of hair against objects such as groom- Modulus extension break extension
ing devices or even other hair is a major (GPs) (MJ/m2) (MJ/m2) (%)
factor in hair damage. The absorbance Indian (Oil user) 2.73 a 0.402 a 1.98 a 51.15 a
values for protein analysis of different
Indian (Non oil user) 2.90 b 0.414 b 1.92 a 49.80 b
ethnic hair are shown in Table 5 and
Fig. 6. Results reveal that protein loss is Values in different letters in superscript are significantly different (p<0.05)
significantly more in African hair fol-
Table 4 Tensile strength of Indian Hair
lowed by Caucasian and Indian hair. This
trend of protein loss studies are as ex-
pected and in line with the surface The Comparison between Oiled and Un- cantly reduces the loss of protein as
analysis (SEM) and porosity measure- oiled Indian hair is presented in Table 6 compared to unoiled hair.
ment. and Fig. 7. As observed oiling signifi-

Absorbance at 750 nm
Hair Type Mean ± SE

Indian 0.0180 ± 0.0027 a

Caucasian 0.0708 ± 0.0030 b

African 0.1734 ± 0.0170 c

Values in different letters in superscript are significantly different (p<0.05)

Table 5 Protein loss from hair in various ethnic groups -Fig. 6 Protein loss for hair from various ethnic groups

Absorbance at 750 nm
Hair Type Mean ± SE

Indian (Oil user) 0.0180 ± 0.0027 a

Indian (Non oil user) 0.0599 ± 0.0079 a

Values in different letters in superscript are significantly different (p<0.05)

Table 6 Protein loss from Indian hair

-Fig. 7 Protein loss from Indian hair

-Fig. 8 Luster values for hair from various ethnic groups -Fig. 9 Luster values for Indian hair

38 SOFW-Journal | 136 | 11-2010


Luster
Indian hair demonstrated more luster
values as compared to Caucasian and
African hairs. The luster value was least
for African hair indicating that African
hair is the most damaged among the
three ethnic groups (Fig. 8).
There was no significant difference in
luster values between hair of Indian oil
users and non oil users (Fig. 9).
 Conclusion
The results of this study were in accor-
dance with the previously published da-
ta on the hair morphology (2). Indian
hair samples showed less porosity and
more luster values than the Caucasian
and African hair samples. Comparisons
between Indian oiled and unoiled sam-
ples suggest that oiling modifies the di-
ameter of hair and lowers loss of vital
proteins. Though oiling does not direct-
ly affect the tensile properties, a favourable
change was seen in the break extension
values. This parameter is important from
grooming perspective since it would mean
that oiled hair would extend more before
breaking during combing as compared to
untreated hair. Further controlled trials
are required to establish the impact of
grooming practices on the physical char-
acteristics of hair.
Acknowledgement
The authors would like to thank Dr. Ram
Ramaprased, TRI Princeton, for his help
and support.
References
(1) Franbourg A, Hallegot P, Baltenneck F,
Toutain C, Leroy F. Current research on ethnic
hair. J Am Acad Dermato. 2003;48:S115-19
(2) Khumalo NP, Doe PT, Dawber RP, Ferguson DJ.
What is normal black African hair? A light and
scanning electron-microscopic study. J Am
Acad Dermatol 2000;43:814-20
(3) Wei G, Bhushan B, Torgerson PM. Nanome-
chanical characterization of human hair us-
ing nanoindentation and SEM. Ultramicros-
copy 2005;105:248-66
(4) Bhushan B, Wei G, Haddad P. Friction and
wear studies of human hair and skin. Wear
2005;259:2591012-1021
SOFW-Journal | 136 | 11-2010
< Content
CO S M ET I C S
HAIR CARE
(5) Kreplak L, Briki F, Duvault Y, et al. Profiling
lipids across Caucasian and Afro-American
hair transverse cuts, using synchrotron in-
frared microspectrometry. Int J Cosmet Sci
2001;23:369-74
(6) Gold RJ, Scriver CG. The amino acid composi-
tion of hair from different racial origins. Clin
Chim Acta 1971;33:465-66
(7) Dekio S, Jidoi J. Amounts of fibrous proteins
and matrix substances in hairs of different
races. J Dermatol 1990;17:62-64
(8) Tate ML, Kamath YK, Ruetsch SB, Weigmann
HD. Quantification and prevention of hair
damage J Soc Cosmet Chem 1993;44:347-71
(9) Mohile RB, Rele AS, Kamath Y. Benefit of Co-
conut Oil Relevance to Hair Damage Part III.
IFSCC 2001;4:277-79
(10) Valko EI, Barnett G. A study of the swelling of
For when
hair in mixed aqueous solvents J. Soc. Cosmet.
Chem. 1952;3:108-17
you want
(11) Menkart J, Wolfram LJ, Mao I. Caucasian hair,
Negro hair and wool: similarities and differ-
ences J Soc Cosmet Chem. 1966;17:769-87
some clarity.
(12) Sandhu S, Robbins C. A simple and sensitive
technique, based on protein measurement to
assess surface damage to human hair. J Soc
Cosmet Chem. 1993;44:163-75
(13) Keis K, Ramaprasad KR, Kamath YK. Studies
of light scattering from ethnic hair fibers. J
Cosmet Sci 2004; 55:49-63
(14) Ruetsch SB, Kamath YK, Rele AS, Mohile RB.
Secondary ion mass spectrometric invstiga-
symbio®solv XC
tion of penetration of coconut and mineral PEG-free natural solubiliser for
oils into human hair fibers: Relevance to hair
damage. J Cosmet Sci 2001;52:169-84
transparent solutions.
(15) Rele AS, Mohile RB. Effect of mineral oil, sun-
flower oil and coconut oil on prevention of
hair damage. J Cosmet Sci 2003;54:175-92
Authors addresses:
* Dr. Yash Kamath
Kamath Consulting Inc
11 Deer Park Drive Suite 206
Monmouth Junction
NJ 088, USA
** Dr. Vaishali Gode, Nitesh Bhalla
Bhargavi Kalghatgi
Dr. Sudhakar Mhaskar
Marico Ltd.
Bombay College of Pharmacy Building
CST Road, Santacruz (E)
Mumbai, India
Corresponding author:
Dr. Vaishali Gode
Email: vaishalig@maricoindia.net intelligence behind beauty

www.dr-straetmans.de

CO S M ET I C S
P R E S E R VAT I V E S
S. Subramanian, A. Wingenfeld*
Fragrance Ingredients with Growth
Inhibiting Activity
 Aromatics with Antimicrobial
Properties
Conarom™ P consists of phenethyl alco-
hol, a nature-identical fragrance addi-
tive in glycolic solution. The aroma com-
ponent is also found in a variety of
plants - rose, carnation, hyacinth and or-
ange blossom. Conarom H-3 is a glycol-
ic blend of Heliotropine and Phenyl-
propanol. The aldehydic component and
one of the glycolic components are also
found in a variety of plants.
The aroma component of the rose-like
Conarom™ P or the vanilla-like Conarom™
H-3 may be used to complement the ex-
isting fragrance of formulations. An
added feature of these nature-identical
ingredients in formulation is that they
may be labeled simply as parfum or as
aroma in end-use packaging. Alterna-
tively, they can be listed by their INCI.
Included among the benefits of Cona-
rom P and Conarom H-3 is broad-spec-
trum inhibition of gram-positive and
gram-negative bacteria, yeast and mold.
Both products can be used in a pH range
of 4 – 8 and show good compatibility in
several leave-on and rinse-off personal
care formulations.
Many of the natural or nature-identical
essential oils contain ingredients which
are known as Fragrance allergens and are
listed in the European Cosmetics Direc-
tive. Keeping this in mind, ISP has care-
fully formulated its Conaroms, such that
the ingredients in the Conarom range do
not contain any of the twenty-six so-
called Fragrance allergens listed in An-
nex III of the European Cosmetics Direc-
tive (76/768/EC).
40
< Content
Benefits of Conarom P and Conarom H-3
• Aromatics with antimicrobial
properties
• Broad spectrum efficacy
• Broad pH range 4 – 8
• Can tolerate high temperatures
• Easy-to-handle liquid
• Can be used in leave-on and rinse-off
applications
Introduction
S
ome consumers actively re-
quest alternatives to tradi-
tional preservatives. In re-
sponse to these requests, mar-
keters and formulators are asking
suppliers to dig deep into their
toolboxes and present novel yet
viable preservation strategies. ISP
fulfils these requests with aromat-
ics that inhibit the growth of mi-
croorganisms. Used in a wide
range of leave-on and rinse-off
applications, Conarom™ P and
Conarom™ H-3 can be labeled as
parfum or aroma in end-use pack-
aging. This article outlines the ap-
plications and benefits of the
Conarom range and presents the
results of well-preserved personal
care formulations.
Formulation Guidelines
When formulated into products at am-
bient temperature, the Conarom range
can be added to any phase of the pro-
duction process. In emulsions, Conarom P
and H-3 can be added to the aqueous
phase up to 80 °C with adequate mixing,
or preferably to the finished emulsion. In
aqueous products with low levels of
emulsifiers, intensive mixing is recom-
mended to achieve uniform dispersion.
The products are recommended for use
at concentrations ranging from 0.3% to
2.0%. Note that strong oxidants and
strong alkalis can lead to decomposition
of the active. Nonionic surfactants such as
Polysorbate 80 may cause deactivation.
 Preservative System Efficacy in
Personal Care Formulations
To evaluate the efficacy of these mate-
rials, challenge testing was conducted in
several formulations using the modified
PCPC method.
This challenge test is a 28-day program
used to verify the effectiveness of a
preservative system in a finished person-
al care formulation. Select personal care
formulations were inoculated with mi-
croorganisms as pure cultures at the on-
set of testing (0 hours), then sampled at
48 hours and 7-day intervals for 28 days.
At 21 days, the formulations were re-in-
oculated with the same microorganisms.
Pass/fail criteria were based on modified
PCPC protocol.
The formulation and challenge test data
along with the challenge test procedures
are detailed below.
SOFW-Journal | 136 | 11-2010
 < Content

Global Ingredients &

Formulations Guide 2010
The Green Book of Cosmetics

– Systems for an Easier Choice and Use of Natural Ingredients

– Certifying Bodies
– Ingredients of Natural Origin with Certificate
– Ingredients of Natural Origin
– Natural Perfume
– Test Institutes

Environmental care and sustainability is an ever increasing challenge for cosmet-

ic production. In a world where nature becomes endangered by global pollu-
tion and climate change the human nature turns toward preserving and protect-
ing nature. Ecologic – economic – sustainable – safe – effective; these are the
requirements for cosmetic products today.
Cosmetics become »green«. More and more manufacturers make an effort for
ecologically sustainable raw materials. They should be natural, go easy on re-
sources, renewable and biodegradable. Nevertheless may one’s sights be low-
ered in use, efficiency and stability of cosmetics. In all areas of raw materials
»green innovations« are already offered, even where it couldn’t be expected for
a long time. In order to prove the naturalness proposition of the »green raw ma-
terials«, more and more »green labels« developed. The green trend requires
additionally efficacy tests that support the new product generation with adver-
tizing effective claims. The new »green raw materials«, with a variety that for
the formulator is not easily manageable any more, are nowadays even catego-
rized by one or the other supplier, to facilitate the choice. In the present »green
book of cosmetics« the reader will find beside the attractive »green« formula-
tion part the well known company index and the supplier profiles. One can con-
vince himself, how many creative and innovative ideas for the use of natural
materials in cosmetics are already implemented. This process just begun, let us
look forward to many more ideas from nature.

• 350 Pages • 11th Edition • German Version also available

ORDER FORM Order-No. 500-1060

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Global Ingredients & Formulations Guide 2010 – Content

Systems for an Easier Choice Natural Deodorant Concepts: Limitations and Possibilities
and Use of Natural Ingredients (Fernando Ibarra)
A Successful Combination –
Naturalness and Performance Symbiomuls WO: A Natural and Easy-to-Use WO-Emulsifier
(Fernando Ibarra)
(Ulrich Isseberner)

Naturals and Their Historical Role in Traditional Vanatural® XGB: Smectite-Xanthan Gum Hybrid
Medicinal Systems – Emulsion Stabilizer
(Peter Ciullo)
Resulting in Valuable Cosmetic Ingredients for
Modern Applications
(Martina Axterer, Cornelia Jones)
Expressions of Differential Genes involved in the Maintenance
of Water Balance in Human Skin by Piptadenia Colubrina Extract
(Maria del Carmen Velazquez Pereda, Gustavo de Campos Dieamant,
Certifying Bodies
Samara Eberlin, Rejane Maria Werka, Débora Colombi, Mary Luci de
Certifying Wild-Collected Cosmetics Ingredients Souza Queiroz, Luiz Claudio Di Stasi)
(Michelle Arts)
Algae Extracts and Ingredients in Cosmetic Products
The NATRUE Label – An Overview (Hans-Peter Hanssen)
(Vincent Letertre)
Emulsense – The New Era of Natural Cationics
Ingredients of Natural Origin (Martina Heldermann)
with Certificate
The Use of Organic Pracaxi Oil (BERACARE BBA) in When it Comes to Feel, is Your Cream the Real Deal?
Substitution to Synthetic Cationic Surfactants in Special Sensory Properties – Don’t be Deceived
Hair Conditioning Studies (Nicole Häusler)
(Cíntia Baradel)
Actives of Magnolia Bark to Stop Inflamm-Aging in the Skin
Integrating Natural, Sustainable and Performance (Daniel Schmid, Irene Montano)
Characteristics in Personal Care Products
(Brajesh Jha, Hürgen Meyer, Gabriele Polak) Efficacy of the Application of Botanical Extracts on Human Hair
(Aurora Benaiges, Blanca Martínez-Teipel, Ricard Armengol,
Natural Hyaluronic Acids and Their Applications Clara and Coderch Luisa Barba)
in Dermocosmetics
(Vera Maienschein) New Nanocapsules with High Loading of UV-Filters
(Gabriele Blume, Katinka Jung)
Natural Cosmetics: Use of »Sigesbeckia Orientalis
Extract« to Boost the Efficacy of Preservative Systems Natural Perfume
(Andrea Mitarotonda, W. Jermyn, N. Williams O’Hanlon) The Magic of Natural Creativity
(Claudia Valder)
Cayoma® Olive the Active Ingredient for Skin Whitening
and Age Spot Reduction Test Institutes
(Maria Lüder, Joachim Blank) Innovative Methods to Reveal the Full-Spectrum
Potential of Natural Cosmetics
Development of »Certified« Organic Active Ingredients, (Peter Girling, Margret Ebauer)
More than a Concept...
(David Boudier, Catherine Lenaers, Claire Sabbadini, Formulations
Delphine Creel, Brigitte Closs)
Body Care, Lip Care, Eye Care, Skin Care, Personal Care,
Hair Care, Disclaimers
Sourcing Shea Butter in 2010:
A Sustainability Check
Company Index
(Peter Lovett)
Contact Addresses, Company Description, Product Range, Service
Ingredients of Natural Origin
Subject Index
An Extract Derived from Broccoli Sprouts to
Regulate the Synthesis of Endogenous
Antioxidants and Detoxify Problematic Metabolites Suppliers’ Directory
(Walter P. Smith)

Exotic Butters for New Sensorial and Marketing

Dimensions
(James J. Ramirez, Larry S. Moroni) www.sofw.com
 < Content
CO S M ET I C S
P R E S E R VAT I V E S

Phase Ingredients Trade Name % w/w

A Deionized Water 45.31

Aminomethyl Propanol AMP-95 (1) 0.02
Acrylic Acid/VP Crosspolymer UltraThix™ P-100 (2) 1.00
Aminomethyl Propanol AMP-95 (1) 0.44
B Deionized Water 36.00
Polyquaternium–28 Conditioneze® NT-20 (2) 9.00
C Deionized Water 4.24
Sodium Hydroxide (10% Aq. Soln.) 0.56
PVM/MA Copolymer Gantrez® S-97 BF Polymer (2) 0.20
D Cyclopentasiloxane Si-Tec™ CM 40 (2) 1.50
Dimethicone and Dimethiconol Si-Tec™ DM 3096 (2) 0.10
E Parfum Conarom P (2) 1.00
Caprylic/Capric Triglyceride (and) Cymbidium Grandiflorum Orchid Complex OS (2) 0.50
Flower Extract
F Mica (and) Titanium dioxide Timica Silkwhite 110W (3) 0.13
Total 100.00

(1)
Angus (2) ISP (3) Engelhard

Procedure:
1. To make Phase A first charge water and first increment of AMP-95 into main container and mix with propeller agitation.
2. Disperse UltraThix™ P-100 into water with vigorous propeller agitation sufficient to create a vortex. Mix for 30 minutes.
3. Add second increment of AMP-95 and mix until uniform.
4. Add water of Phase B into a premix container and mix with moderate propeller agitation. Add Conditioneze® NT-20 and
mix until uniform.
5. Add water of Phase C into a separate premix container and mix with moderate propeller agitation.
6. Add sodium hydroxide solution to Phase C and mix until uniform.
7. Sprinkle Gantrez® S-97 BF Polymer into vortex of Phase C and mix until uniform. Adjust phase C to pH 6.96 ± 0.05 with
sodium hydroxide solution.
8. Increase agitation of contents of Phase B (~ 1000 rpm). Add Phase C to Phase B over the course of 20-30 seconds.
Mix with fast propeller agitation for ten minutes.
9. Add combined Phases B and C to Phase A. Mix until uniform.
10. Premix Phase D, add to main batch and mix until uniform.
11. Add ingredients of Phase E to main batch and mix until uniform.
12. Add Phase F to main batch and mix until uniform

Formulation FIT-ENDS-Split End Mending Reconstructor Cream

Inoculum concentration (CFU/g)

Organism 0 Hours 21 Days

Staphylococcus aureus 3.9x106 3.0x106

Escherichia coli 9.2x106 7.4x106
Pseudomonas aeruginosa 5.6x106 3.2x106
Burkholderia cepacia 3.1x106 3.4x106
6
Candida albicans 6.0 10 1.6x106
Aspergillus brasiliensis 7.0x105 8.0x105

SOFW-Journal | 136 | 11-2010 43

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CO S M ET I C S
P R E S E R VAT I V E S

Preservative efficacy test results (CFU/g)

Organism 48 Hours 7 Days 14 Days 21 Days 28 Days

Staphylococcus aureus <10 <10 <10 <10 <10

Escherichia coli <10 <10 <10 <10 <10
Pseudomonas aeruginosa <10 <10 <10 <10 <10
Burkholderia cepacia <10 <10 <10 <10 <10
Candida albicans <10 <10 <10 <10 <10
Aspergillus brasiliensis <10 <10 <10 <10 <10

Phase Ingredients Trade Name % w/w

A Deionized Water 61.00

Propylene Glycol (1) 3.00
Kaolin BC 2457 (2) 2.00
Vinyl Caprolactam/VP/Diemthylaminoethyl Advantage® S (3) 1.00
Methacrylate Copolymer
PVP PVP K-30 (3) 1.00
B Lanolin Wax Protalan Wax (4) 8.00
Beeswax Refined Yello Beeswax Prills (5) 10.00
Cetearyl Alcohol (and) Behenyl Alcohol (and) Hydroxyethyl Prolipid® 161 (3) 3.00
Cetearamidopropyldimonium Chloride
Glyceryl Stearate (and) PEG-100 Stearate Arlacel 165 (6) 2.50
Quaternium-70 (and) Propylene Glycol Ceraphyl® 70 (3) 0.50
Octyldodecyl Stearoyl Stearate Ceraphyl® 847 (3) 2.00
Shea Butter Refined Shea Butter (3) 5.00
C Parfum Conarom™ H-3 (3) 1.00
Total 100.00

(1)Rita (2) Brenntag (3) ISP (4) Protameen (5) Frank B. Ross (6) Croda
Procedure:
1. Add water of Phase A to main container and mix with fast sweep agitation producing a vortex.
2. Add Propylene Glycol and polymers and mix until uniform. Sprinkle Kaolin and mix until dispersed.
3. Heat batch to 75 °C. Premix Phase B by melting ingredients to 80 °C. Add Phase B to A, while mixing with sweep agitation.
4. Start cooling the batch. As the batch thickens, increase sweep mixing speed.
5. Add preservative at 65 °C. Fill into jars at 60 °C.
Formulation Still in Bed Clay

Inoculum concentration (CFU/g)

Organism 0 Hours 21 Days

Staphylococcus aureus 6.0x106 2.9x106

Escherichia coli 4.7x106 4.9x106
Pseudomonas aeruginosa 4.3x106 3.8 x106
Burkholderia cepacia 6.0x106 2.9x106
6
Candida albicans 4.0 10 2.9x106
Aspergillus brasiliensis 7.0x105 7.0x105

44 SOFW-Journal | 136 | 11-2010

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CO S M ET I C S
P R E S E R VAT I V E S

Preservative efficacy test results (CFU/g)

Organism 48 Hours 7 Days 14 Days 21 Days 28 Days

Staphylococcus aureus <10 <10 <10 <10 <10

Escherichia coli <10 <10 <10 <10 <10
Pseudomonas aeruginosa <10 <10 <10 <10 <10
Burkholderia cepacia <10 <10 <10 <10 <10
Candida albicans <10 <10 <10 <10 <10
2
Aspergillus brasiliensis 2.2x10 <10 <10 <10 <10

 Preservative Efficacy Study

Comparing Two Ingredients

To evaluate the efficacy of the aromatic

ingredient, Conarom P, against a com-
monly used preservative system of
Phenoxyethanol-Caprylyl Glycol (trade
name: Optiphen), a repetitive challenge
test was conducted. Both Conarom P
and Optiphen were tested at two differ-
ent use levels: 1.0% and 1.5%.
An unpreserved personal care formula-
tion was inoculated with a bacterial and
fungal composite at the start of the
study (0 hours). The inoculations were re-
peated three times at an interval of 14
days. The initial observation was com-
pleted 1 week after the first inoculation.
After the fourth inoculation, additional
viable counts were performed in shorter
time intervals to determine the kill-rate
of the investigated preservation systems.
While the unpreserved control remained
highly susceptible to bacteria, yeast and
-
Fig. 1 Preservative efficacy study results
mold, the study results showed Conarom
P has the same effective kill-rate as the
Optiphen system. Both systems killed the
microbial composite by Day 2. No further used to help preserve a wide range of at Wayne, NJ (USA) and Memmingen,
growth was seen through the duration of leave-on and rinse-off formulations, in- Germany respectively. The authors also
the study, indicating that Conarom P cluding sunscreens, serums, shower gels, thank Jadwiga Karolak and Jose Mar-
may serve as a viable alternative to com- creams, lotions, shampoos, conditioners tinez-Santiago for the formulation work.
monly used antimicrobial systems. and styling applications. Available as al-
Fig. 1 shows the results of Conarom P ternatives to traditional preservative sys-
and Optiphen at 1.0% use level. tems, the Conarom range is efficacious in * Authors’ address:
formulations between 0.3% - 2.0% use Sangeetha Subramanian
level. Andrea Wingenfeld
 Conclusion ISP Personal Care
1361 Alps Road
The Conarom range offer mild aromatic Acknowledgement Wayne NJ 7470
benefits in addition to antimicrobial func- The authors would like to thank the ISP USA
tionality. The aroma component comple- microbiological department led by Susan Email: ssubramanian@ispcorp.com
ments the existing fragrance while the Lindstrom and Tanja Schubert for con- awingenfeld@ispcorp.com
antimicrobial activity can be effectively ducting the preservative efficacy testing


SOFW-Journal | 136 | 11-2010 45


CO S M ET I C S
ST E M C E L L S
L. Cattuzzato, C. Sanchez, A. De Pooter, M. Puginier, S. Dumont*
Complementary In Vitro Models to Investigate
the Mode of Action of Active Ingredients on the
Protection of Extracted Epidermal Stem Cells
Against Different Types of Stresses
 Introduction
Epidermal stem cells (ESC) are necessary
for epidermis renewing. These cells are
also known to be more or less resistant
to different types of apoptosis, in com-
parison with their keratinocyte counter-
parts: UV-induced cell death (1), oxida-
tive stress-induced apoptosis (2) or anoïko-
sis (a particular apoptosis resulting from
loss of adhesion to extracellular matrix)
(3). Thus, some models have been devel-
oped to extract ESC from epidermis, to
culture them and to investigate their
specific functions such as colony-form-
ing capacity or their sensitivity to oxida-
tive stress (2).
Survivin, an anti-apoptotic protein, has
been shown to be specifically expressed
in the nucleus of ESC, among an ex-
tracted population composed of ESC,
transitory-amplifying cells and differen-
tiated keratinocytes (4). ΔNp63 is a p53
anti-apoptotic homologue which has al-
so been shown to be specifically ex-
pressed by ESC (5). It has been described
to give UVB-resistance to ESC and to de-
crease in these cells after such a stress
(6). The NGF-pathway has also been
shown to be implicated in this resistance,
inducing an increase in the production
of the anti-apoptotic Bcl-2 molecule (1,
7). Other ESC-associated markers have
been described, such as MCSP (Melanoma
Chondroitin Sulfate Proteoglycan) (8, 9),
or a high level of β1-integrin expression
(3).
The purpose of this study was, first, to
develop new in vitro models to investi-
gate ESC response after these different
types of stresses and, second, to investi-
gate the preventive effect of active in-
46
< Content
gredients. We particularly focused on
Cocoyl alanine (CA), an antioxidant and
anti-wrinkle cosmetic product. Indeed,
previously obtained results suggested
that it was able to protect elderly skin
explants from ex vivo culture-induced
decrease in survivin expression (nuclear
form) within epidermis (10).
 Methodology
Reagents and materials
Cocoyl Alanine (CA) was manufactured
by Seppic. α-tocopherol was purchased
from BASF. Foetal Beef Serum (FBS), hu-
man fibroblasts, Modified Eagle’s Medi-
um (MEM), Ham’s F12 medium, M199
medium, L-glutamin, penicillin-strepto-
mycin, gentamycin, trypsin/EDTA and
sodium bicarbonate were all purchased
from Lonza. Phosphate Buffered Saline
(PBS) was purchased from Biomérieux.
Dubelcco’s Modified Eagle’s Medium
(DMEM), hydrocortisone, transferrin, in-
sulin, EGF, bFGF, mitomycin C, Hoechst
reagent, dispase, DMSO, methanol, MTT,
collagen IV and H2O2 were all purchased
from Sigma-Aldrich. Mouse monoclonal
antibodies against human β1-integrin,
against human MCSP and against hu-
man ΔNp63 were purchased from Tebu.
Alexa Fluor568-conjugated secondary goat
anti-mouse antibody was purchased
from Invitrogen/Molecular Probes. Mouse
monoclonal neutralizing antibody against
human β1-integrin was purchased from
Beckman Coulter. FragEL™ DNA fragmen-
tation detection kit and K252a were pur-
chased from Calbiochem. Acetone was
purchased from Xilab. Hema- toxylin and
eosin were purchased from Labonord.
ESC extraction and culture
ESC were isolated from epidermal cell
suspensions, themselves extracted from
healthy patient abdominal skin explants
after standard dispase and trypsin/EDTA
procedures. ESC were isolated by select-
ing the keratinocytes which were able to
adhere on type IV collagen-precoated
dishes in 5 min., as previously described
(11, 12). ESC were cultured on post-mi-
totic fibroblasts at 37 °C in a humidified
incubator under a 5% CO2, 95% air at-
mosphere. ESC medium was composed of
DMEM/F12 (3:1) completed with 0.4 µg.
mL-1 hydrocortisone, 5 µg.mL-1 transfer-
rin, 5 µg.mL-1 insulin, 10 ng.mL-1 bFGF,
10 ng.mL-1 EGF, 100 µg.mL-1 streptomycin,
100 U.mL-1 penicillin and 10% FBS. Medi-
um was replaced every two or three days.
ESC were used at passage P2 or P3 and
seeded in 48-microwell culture plates.
Medium was replaced every two days.
Investigation of ESC phenotype by
immunofluorescence
After removal of culture medium, cells
were rinsed and fixed with a frozen so-
lution of acetone/methanol (8/2, v/v)
then rinsed with a solution of PBS/ Mon-
tanox™ 20 0.05% (wash buffer). Non spe-
cific sites were saturated with a 10%
nonfat dry milk solution. Cells were then
incubated with the appropriate primary
antibody dilution for 1 hour at room tem-
perature (anti-MCSP: 1:100, anti-β1-in-
tegrin: 1:100). At the end of the incuba-
tion, cells were washed and incubated
with the Alexa Fluor-conjugated sec-
ondary antibody dilution for 1 hour at
room temperature in the dark. Cells were
then washed again and incubated with
SOFW-Journal | 136 | 11-2010
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CO S M ET I C S
ST E M C E L L S
the Hoechst dilution (2 µg.mL-1) for 15
min. at room temperature in the dark.
After washing, microscopical observa-
tions were performed with an inverted
microscope equipped with UV epifluo-
rescence.
Investigation of ESC resistance to UVB
On day 5 after seeding, cells were treat-
ed or not with K252a (200 nM). On day
7, ESC were irradiated or not with dif-
ferent doses of UVB rays (i.e. 25 to 200 mJ.
cm-2). At the end of the irradiation, cells
were further cultivated in the ESC medi-
um for 24 hours at 37 °C before the in-
vestigation of cell viability by a standard
MTT procedure.
Investigation of oxidative stress on ESC
culture and apoptosis
On day 4 after seeding, cultures were
incubated or not with CA (0.00001%,
0.0001% or 0.001%, cp) or with 10 µg.mL-1
α-tocopherol, both diluted in ESC medi-
um containing only 2% FBS, for 24h. On
day 5, cultures were treated with 50 µM
H202 for 18h. After this period, medium
was replaced by 2% FBS-ESC medium
and cells were further cultured for 48h.
Cultures were then rinsed with PBS and
cells were fixed with a frozen solution of
acetone/methanol (8/2, v/v). Finally, cells
were rinsed either with PBS before per-
forming a standard hematoxylin-eosin
(HE) staining or with TBS before detec-
Hoechst
ESC
HEK
-Fig. 1 Investigation of ESC-related markers by immunofluorescence
48
< Content
tion of DNA fragmentation with the
FragEL™ DNA fragmentation detection
kit, according to the instructions provid-
ed by the manufacturer.
Investigation of UVB on ESC ΔNp63
level and apoptosis
On day 4 after seeding, cultures were
incubated or not for 72h with CA
(0.00001%, 0.0001% or 0.001%, cp) or
with 20 U.mL-1 IL-1 β. After this period,
medium was replaced and cells were fur-
ther cultured for 18h. ESC were then ir-
radiated or not with 200 mJ.cm-2 UVB
rays. At the end of the irradiation, cells
were further cultivated in the ESC medi-
um for 24 or 48 hours. For experiments
related to the investigation of the NGF
pathway, the inhibitor K252a (200 nM)
was added to the 2% FBS-ESC medium.
In this case, cells were cultivated 48h be-
fore and after irradiation with a lower
dose of UVB rays (50 mJ. cm-2). In both
kinds of experiments, cultures were then
rinsed with PBS. Cells were fixed with a
frozen solution of acetone/methanol
(8/2, v/v). Finally, cells were rinsed either
with PBS before performing an anti-
ΔNp63 immunoflorescent staining (24h
post-irradiation) or with TBS before de-
tection of DNA fragmentation.
Investigation of AB1I on ESC anoïkosis
ESC were seeded in 6-microwell culture
plates. Medium was replaced every two
β1-integrin Hoechst
days. On day 4, cultures were incubated
or not for 72h with CA (0.00001%,
0.0001% or 0.001%, cp). After this peri-
od, cells were dissociated by standard
trypsin/EDTA procedure then incubated
or not during 2h with mouse monoclon-
al neutralizing antibody against human
β1-integrin (1:250). After the end of the
incubation, cells were centrifugated and
the cellular pellets were resuspended in
formaldehyde 4%. After 48 hours at 4 °C,
fixed cells were deposited on slides and
rinsed with TBS before detection of DNA
fragmentation.
Image analysis, data analysis and
statistical analysis
All experiments were performed in trip-
licates with at least four samples per ex-
periment.
Photographs of each condition were tak-
en with the associated numeric camera.
Pictures were analysed with the NIS-Br
software. ESC colony size was calculated
by the software by measuring the area of
colony and expressed as µm2. Propor-
tions of positive cells for a given marker
(or for apoptosis) were calculated by re-
porting the number of positive cells to
that of total ESC nuclei (Hoechst+ cells)
and expressed as percentages. For each
group, the mean and the standard devi-
ation were calculated, reported to the
control group and expressed as variation
percentages. Statistical significance was
assessed using an ANOVA test followed
MCSP
SOFW-Journal | 136 | 11-2010
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CO S M ET I C S
ST E M C E L L S

by a Bonferroni adjustment, with p<0.05
being considered significant (compared
with the stress condition). In all cases, a
percentage of restoration was calculat-
ed. Finally, mean effects were calculated
on the basis of results which were sig-
nificant in each experiment.
 Results
Investigation of ESC phenotype
Extracted and cultured cells expressed
the ESC-related marker MCSP (Melano-
Investigation of H2O2 stress on ESC
colony size and apoptosis
H2O2 treatment (50 µM) of cultures in-
duced a significant decrease in the size
of ESC colonies, i.e. of 58% (Fig. 2). It al-
so induced a significant increase in the
proportion of TUNEL-positive (TUNEL+)
cells, i.e. of 284% (Fig. 3). Preliminary re-
sults concerning the investigation of the
proportion of survivin+ positive cells al-
so showed a decrease after H2O2 treat-
ment (-65%, DNS).
The reference molecule α-tocopherol
(10 µg.mL-1) partially limited such effects,
inducing a restoration effect of 94% and
68%, regarding the size of ESC colonies
(Fig. 2) and the proportion of TUNEL+
cells (Fig. 3), respectively. It also induced
a preventive effect (37%) against the
H2O2-induced decrease in the proportion
of survivin+ positive cells (DNS).
CA tested at the two higher concentra-
tions (0.0001% or 0.001%), also showed
a restoration effect regarding both the
size of ESC colonies, respectively of 54%
and 69% (Fig. 2), and the proportion of
TUNEL+ cells, respectively of 32% and
54% (Fig. 3). It also induced a preventive

Control (A) H2O2 (B) α-tocopherol (C) CA 0.001% (D)

-Fig. 2 Investigation of colony size by standard HE staining in ESC cultures after oxidative stress.
A: control (non treated cultures), B: 50 µM H2O2, C: 50 µM H2O2 + 10 µg.mL-1 α-tocopherol, D: 50 µM H2O2 + 0.001% cocoyl
alanine

ma Chondroitin Sulfate Proteoglycan)

and showed a high level of β1-integrin
expression, in comparison with normal
human keratinocytes (NHK) (Fig. 1).
Investigation of ESC resistance to UVB
Both NHK and ESC were submitted to a
range of UVB ray doses to evaluate their
resistance to such a stress. Mortality in-
duced by UVB did not exceed 11% in ESC,
even when the cells were exposed to the
highest dose of UVB, i.e. 200 mJ.cm-2 (da-
ta not shown, DNS). On contrary, cell vi-
ability was highly impaired in NHK, as
from the dose of 50 mJ.cm-2 (DNS). When
pre-treated with K252a, ESC viability
was decreased to 70% as from the dose
of 25 mJ.cm-2 (DNS), while the differ-
ences observed between untreated and
K252a-treated NHK were not significant -Fig. 3 Investigation of the level of apoptosis in ESC cultures by TUNEL after oxida-
(DNS). tive stress.
Thus, two UVB doses were chosen: 200 mJ. A: control (non treated cultures), B: 50 µM H2O2, C: 50 µM H2O2 + 10 µg.mL-1 α-
cm-2 for the investigation of ESC resis- tocopherol, D: 50 µM H2O2 + 0.0001% cocoyl alanine, E: 50 µM H2O2 + 0.001% co-
tance in basal conditions; and 50 mJ.cm-2 coyl alanine. Statistical analysis performed using ANOVA test followed by a Bon-
for the investigation of ESC resistance in ferroni adjustment compared with the H2O2 condition (B): LS0.05<p<0.1 ; **p<0.01
conditions of NGF pathway inhibition ; ***p<0.001. X% = restoration percentage
(i.e. treatment with K252a).

SOFW-Journal | 136 | 11-2010 49

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CO S M ET I C S
ST E M C E L L S

effect (from 37% to 52%) against the

H2O2-induced decrease in the proportion
of survivin+ positive cells (DNS).

Investigation of UVB stress on ΔNp63

level
UVB irradiation (200 mJ.cm-2) of ESC in-
duced a significant decrease in the pro-
portion of ΔNp63+ cells, i.e. of 67%
(Fig. 4).
The reference molecule IL-1β (20 U.mL-1)
partially limited such effects, inducing a
restoration effect, i.e. of 58%, regarding
the proportion of ΔNp63+ cells (Fig. 4).
CA tested at the two higher concentra-
tions (0.0001% or 0.001%), also showed
a restoration effect regarding the pro-
portion of ΔNp63+ cells, of 29% and
48% respectively (Fig. 4).
-Fig. 4 Investigation of ΔNp63 expression in ESC cultures after UVB irradiation.
A: control (non irradiated cultures), B: 200 mJ.cm-2 UVB, C: 200 mJ.cm-2 UVB + 20
U.mL-1 IL-1β, D: 200 mJ.cm-2 UVB + 0.0001% cocoyl alanine, E: 200 mJ.cm-2 UVB
Investigation of the involvement of the
+ 0.001% cocoyl alanine. Statistical analysis performed using ANOVA test followed
NGF pathway in the UVB-induced
by a Bonferroni adjustment compared with the UVB condition (B): LS0.05<p<0.1;
apoptosis
*p<0.05 ; **p<0.01 ; ***p<0.001. X% = restoration percentage
When ESC were both treated by K252a
and UVB, an increase in the proportion
of TUNEL+ cells was observed from an gether and regarding both phenotype the first model, the protective role of
UVB dose of 50 mJ.cm-2 to 200 mJ.cm-2 and function, these results showed that CA was compared with that of α-toco-
(data not shown). At 50 mJ.cm-2, varia- the extracted cell population was actu- pherol, a well-known antioxidant mole-
tions were of +116% (Fig. 5). ally mainly composed of ESC. cule which had been previously shown to
CA tested at the two higher concentra- Then, three models have been developed protect ESC from oxidative stress (2). As
tions (0.0001% or 0.001%) showed a to investigate apoptosis of these cells. In expected, and in accordance with its pre-
restoration effect regarding the propor-
tion of TUNEL+ cells of respectively 42%
and 52% (Fig. 5).

Investigation of the effect of anti-β1-

integrin antibody on ESC anoikis
AB1I treatment (1:250) of cultures in-
duced a significant increase in the pro-
portion of TUNEL+ cells, i.e. of 203%
(Fig. 6).
CA tested at the highest concentration
(0.001%), showed a restoration effect re-
garding the proportion of TUNEL+ cells,
i.e. of 69% (Fig. 6).

 Discussion
As expected, extracted and cultured cells
expressed the ESC-related marker MCSP
and showed a high level of β1-integrin
expression. Furthermore, the extracted
and cultured cells showed a particularly
high resistance to UVB rays, as described
in literature for ESC (1). Thus, taken to-
-Fig. 5 Investigation of the level of apoptosis in ESC cultures by TUNEL after UVB
irradiation and inhibition of the NGF pathway.
A: 50 mJ.cm-2 UVB, B: 50 mJ.cm-2 UVB + 200 nM K252a, C: 50 mJ.cm-2 UVB + 200 nM
K252a + 0.0001% cocoyl alanine, D: 50 mJ.cm-2 UVB + 200 nM K252a + 0.001%
cocoyl alanine. Statistical analysis performed using ANOVA test followed by a Bon-
ferroni adjustment compared with the UVB+K252a condition (B): ***p<0.001. X%
= restoration percentage

50 SOFW-Journal | 136 | 11-2010


-Fig. 6 Investigation of the level of apoptosis in ESC cultures by TUNEL (fluores-
cence-based protocol) in the anoikis model (statistical analysis performed on the
basis of image analysis values).
A: control (non treated cultures), B: 1:250 AB1I, C: 1:250 AB1I + 0.001% cocoyl
alanine. Statistical analysis performed using ANOVA test followed by a Bonferroni
adjustment compared with the AB1I condition (B): *p<0.05 ; ***p<0.0001. X% =
restoration percentage
viously described antiradical activity
(12), CA did protect ESC against H2O2-in-
duced apoptosis, as illustrated by the
restoration effect that it showed regard-
ing both the size of ESC-colonies and the
proportion of TUNEL+ cells.
In the second model, i.e. UVB model, the
protective role of CA was compared with
that of interleukin (IL)-1β, which is known
to increase the secretion of NGF, pro-
tecting thus cells from apoptosis (13). CA
exerted a protective effect against UVB-
induced decrease in ESC proportion, since
it could partially limit the decrease in
the proportion of ΔNp63+ cells. Such a
property could be due, at least partially,
to the antiradical property of CA. Whether
such protective roles were mediated by
the Nerve growth factor (NGF)-signalling
pathway or not was investigated by de-
creasing the level of UVB irradiation
(50 mJ. cm-2). As expected, in these con-
ditions, inhibition of the NGF-signalling
pathway (using a Tyrosine kinase in-
hibitor, i.e. K252a) was necessary to in-
duce an increase in apoptosis in the ESC
population (TUNEL+ cells) (1, 7). The fact
that CA could also limit UVB-induced
variations suggests that it could partial-
ly restore the NGF pathway. Investiga-
tion of ΔNp63 expression in such condi-
SOFW-Journal | 136 | 11-2010
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tions will have to be performed in order
to confirm these properties and to un-
derstand better the mode of action of
CA.
Finally, in the third model of AB1I-in-
duced anoïkosis, CA could also exert a
preventive effect since it limited the in-
crease in TUNEL+ cells. Thus, these results
suggest that CA could act against apop-
tosis events which would be induced by
a loss of extracellular matrix adherence.
Taken together, these data suggest that
the preventive effect of CA observed in
explant cultures, regarding survivin ex-
pression within epidermis, is likely to be
related to its ability to act against dif-
ferent kinds of stresses on ESC.
 Conclusion
In conclusion, after confirmation of ESC
phenotype (i.e. MCSP+ β1-integrinhigh
ΔNp63+ Surv+) and their functionality
(UVB-resistance), three different kinds of
models could be validated to evaluate
the effect of oxidative stress, UV and loss
of adhesion to extracellular matrix on
ESC apoptosis. Such models also enable
the study of the protective ability of cos-
metic active ingredients. More particu-
CO S M ET I C S
ST E M C E L L S
larly, CA showed a protective effect in
the three aforementioned models, which
is coherent with its antiradical properties
and its clinically proven anti-age activi-
ty. In the future, intracellular signalling
pathways regulating such effects could
be more precisely investigated by study-
ing phosphorylation events and tran-
scription factor activities.
References
(1) Marconi et al. The Journal of Investigative
Dermatology, 113 (1999) 920-927
(2) Noblesse et al. Congrès Annuel de Recherche
Dermatologique de Toulouse, 2008, pp. 24
(3) Tiberio et al. C. FEBS Letters, 524 (2002) 139-
144
(4) Marconi et al. Stem Cells, 25 (2007) 149-155
(5) Kim et al. J. Cutan. Pathol., 36 (2009) 825-
830
(6) Liefer et al. Cancer Research, 60 (2000) 4016-
4020
(7) Marconi, et al. The Journal of Investigative
Dermatology, 121 (2003) 1515-1521
(8) Legg, et al. Development, 130 (2003) 6049-
6063
(9) Ghali et al. The Society for Investigative Der-
matology, 122 (2004) 433-442
(10) Dumont et al. C. International Journal of Cos-
metic Science, 32 (2010) 9-27
(11) Papini et al. Stem Cells, 21 (2003) 481-494
(12) Dong et al. Cell Biology International, 31
(2007) 733-740
(13) Pons et al. European Journal of pharmacolo-
gy, 428 (2001) 365-369
Authors’ :
Laetitia Cattuzzato, Cindy Sanchez Am-
bre De Pooter, Miackael Puginier
Sandy Dumont
SEPPIC Laboratoire de biologie
127 chemin de la Poudrerie
81105 Castres Cedex
France
* Correspondence author:
Email: sandy.dumont@airliquide.com

51

L AU N D RY A N D C L EA N I N G
E U - ECO L A B E L
M. Gast*
Revision of the EU-Ecolabel for Detergents
 Introduction
The EU Ecolabel for environmental-
friendly products is a successful tool for
the promotion of eco-friendly products.
It is part of the common policies for sus-
tainability of production and consump-
tion, with the objective of reducing neg-
ative impacts of consumption and pro-
duction on environment, health, climate
and natu- ral resources. Products with
this ecolabel – the Euro-flower- are sub-
ject to strict impositions in view of the
impact of resources on environment and
health, exceeding legal provisions.
Criteria for the award of the Euro-flower
to laundry detergents were first deter-
mined in 1995. For the first time, a com-
plex point system for ingredients was set
forth for the ecological evaluation in the
scope of international product assess-
ment. It has been discussed amongst ex-
perts and was then ratified by the rele-
vant forums. The proof of cleaning per-
formance guarantees a clean result, be-
cause the suitability of the product for
its claimed purpose must be proven in
a test. Today criteria for the EU Ecolabel
exist for the product groups laundry de-
tergents, dishwashing detergents and
hand dishwashing detergents and all-
purpose and sanitary cleaners.
The criteria for the ecolabel has been re-
viewed in relation to the state-of-art
technology; for laundry detergents and
detergents for dishwashers since 2008
and for hand dishwashing detergents and
all-purpose cleaners since 2009. A mate-
rial aspect of the revision is the substi-
tution of dangerous substances with less
hazardous substances, wherever this is
possible. These new criteria are supposed
52
< Content
to be easily understood and applicable.
They should also consider the newest
technological developments. The ratifi-
cation of the new criteria is planned in
autumn 2010.
However, there is another reason that
made the adjustment of the criteria a ne-
cessity. The Regulation (EC) No 66/2010 of
the European Parliament and the Coun-
cil ratified on 25 November 2009 and in
full force now regulated the basic crite-
ria for the award of ecolabel. This result-
ed in the revision of all existing criteria.
In this context, the most important
change regarding the award criteria is
based on article 6 of the Regulation (EC)
No 66/2010. Article 6 section 6 and arti-
cle 6 section 7 are the most important
items in that context.
Article 6 (3):
EU Ecolabel criteria shall be determined
on a scientific basis considering the whole
life cycle of products. In determining such
criteria, the following shall be considered:
a) the most significant environmental
impacts, in particular the impact on
climate change, the impact on nature
and biodiversity, energy and resource
consumption, generation of waste,
emissions to all environmental;
b) the substitution of hazardous sub-
stances by safer substances, as such
or via the use of alternative materi-
als or designs, wherever it is techni-
cally feasible.
Article 6 (6):
The EU Ecolabel may not be awarded to
goods containing substances or prepa-
rations/mixtures meeting the criteria for
classification as toxic, hazardous to the
environment, carcinogenic, mutagenic
or toxic for reproduction (CMR), in ac-
cordance with Regulation (EC) No 1272/
2008 of the European Parliament and of
the Council of 16 December 2008 on
classification, labelling and packaging
of substances and mixtures nor to goods
containing substances referred to in Ar-
ticle 57 of Regulation (EC) No 1907/2006
of the European Parliament and of the
Council of18 December 2006 concerning
the Registration, Evaluation, Authorisa-
tion and Restriction of Chemicals
(REACh), establishing a European Chemi-
cals Agency.
Article 6 (7):
For specific categories of goods con-
taining substances referred to in para-
graph 6, and only in the event that it is
not technically feasible to substitute
them as such, or via the use of alterna-
tive materials or designs, or in the case
of products which have a significantly
higher overall environment performance
compared with other goods of the same
category, the Commission may adopt
measures to grant derogations from
paragraph 6. No derogation shall be giv-
en concerning substances that meet the
criteria of Article 57 of Regulation (EC)
No 1907/2006 and that are identified
according to the procedure described in
Article 59(1) of that Regulation, present
in mixtures, in an article or in any ho-
mogeneous part of a complex article in
concentrations higher than 0,1% (weight
by weight). Those measures, designed to
amend non-essential elements of this
Regulation, shall be adopted in accor-
dance with the regulatory procedure
with scrutiny referred to in Article 16(2).
SOFW-Journal | 136 | 11-2010
 < Content

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the brand name, a bargain price? Whatever it is, we will work with you to develop
solutions for tomorrow’s successful products. That’s why we already supply more
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 < Content
L AU N D RY A N D C L EA N I N G
E U - ECO L A B E L

As a consequence, any substances en-
dangering health or environment and
exceeding a concentration of 0.01% in a
product are prohibited altogether. This
means that theoretically, ingredients
may not be used if they are found in one
or several of the listed classifications.
In the future, certain ingredients that
cannot be substituted in laundry deter-
gents and cleaning agents must be de-
fined for every product category, for ex-
ample certain tensides or preservatives,
to ensure their continued use.
 State of the Discussion in Regard
to Different Award Criteria
Due to the extent of this document and
the ongoing discussion in view of hand
dishwashing detergents, all-purpose clean-
ers, glass and sanitary cleaners it is not
possible to list all criteria. For laundry de-
tergent and detergents for dishwashers
the discussion has been completed and
the criteria will probably be published in
the official bulletin in February 2011. In
the following you will find a description
of the most important decided or planned
criteria, concerning the use and the eval-
uation of ingredients.
a) Laundry Detergent
When developing the criteria for laundry
detergent, it was also discussed to in-
clude fabric softeners and stain removers
in this paper. However, it was decided to
discuss criteria for fabric softeners in its
own paper. In addition to the inclusion of
stain removers in the product selection,
the following changes are material:
• The use of phosphates is prohibited.
• The amount of chemicals per wash was
reduced from 100 g/wash to 17.0 g/kg
wash for heavy-duty laundry deter-
gent / colour-safe detergent and low-
duty laundry detergents. For stain
removers the value was determined
2.7 g/kg wash.
• New CDV-standards have been set
forth to merge the product evalua-
tion into the current DID-list. For
heavy-duty detergents the CDV will
be 35.000 l/ kg wash in the future and
for low-duty detergents 20.000 l/kg
wash. A CDV of 3.500 l/kg wash was de-
termined for stain removers.

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• There are no ambient standards for
phosphonates.
• New standards have been determined
for the amount of aerobically non-
biodegradable substances (aNBO) and
the amount of anaerobically non-
biodegradable substances (anNBO)
(Table 1).
• Exceptions for the general prohibition
of substances endangering health or
environment have also been decided
(Table 2).
b) Detergents for dishwashers
In addition to the inclusion of liquid rinse
aid in the product selection, the follow-
ing changes are material:
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L AU N D RY A N D C L EA N I N G
E U - ECO L A B E L
• The use of phosphates is prohibited.
• The chemicals used per wash has been
reduced from 22.5 g/wash to TCmax =
20.0 g/wash for single functional dish-
washing detergents and to TCmax =
22.0 g/wash for multi functional dish-
washing detergents.
• New CDV-standards have been set
forth to merge the product evaluation
into the current DID-list. For single
functional dishwashing detergents the
CDV will be 25.000 l/wash in the future,
for multi single functional dishwash-
ing detergents 30.000 l/wash and for
rinse aid 10.000 l/wash.
• There are no ambient standards for
phosphonates.
• New standards have been determined
for the amount of aerobically non-
biodegradable substances (aNBO) and
the amount of anaerobially non-
biodegradable substances (anNBO). For
detergents for dishwashers, the aNBO
will be 1.0 g/wash and the anNBO 5.50
g/wash. For rinse aids the aNBO will
be 0.15 g/ wash and the anNBO will
be 0.50 g/ wash.
• Table 3 shows the exception from the
general prohibition of health endan-
gering substances or substances dan-
gerous for the environment.
c) Hand Dishwashing Detergents
It is foreseen that the area hand dish-
washing detergents will show almost no
COGNIS GmbH, Phone +49 211 79400
care.chemicals@cognis.com, www.cognis.com
 < Content
L AU N D RY A N D C L EA N I N G
E U - ECO L A B E L

changes of the criteria. Apart from the
adjustment to article 6 of the Regulation
(EC) No 66/2010 only the new CDV-stan-
dard is of importance, since the maxi-
mum CDV for hand dishwashing deter-
gents has been reduced from 4,200 to
3,800.
d) All-Purpose and Sanitary Cleaners
Apart from the adjustment to article 6 of
the Regulation (EC) No 66/2010, the de-
finition of all-purpose cleaners for clari-
fying that these products are meant to
be used for the general cleaning of
buildings and the controversial inclusion
of ready-to-use products in the product
selection, the following changes are ma-
terial:
• The maximum standard of VOC will be
• New CDV-standards have been deter-
mined. For all-purpose cleaners, the
maximum acceptable CDV has been
reduced from 20,000 to 18,000, for
window cleaners from 5,000 to 4,800
and for sanitary cleaners from
100,000 to 80,000. Currently, the CDV
standard of 50,000 for ready-to-use
products is controversial.
reduced from 10% to 6%, applicable
to all-purpose cleaners and sanitary
cleaners.

Product type aNBO, powder aNBO, liquid/gel

Heavy-duty laundry detergent, Colour-safe detergent 1.0 g/kg wash 0.55 g/kg wash
Low-duty laundry detergent 0.55 g/kg wash 0.30 g/kg wash
Stain remover (pre-treatment only)* 0.10 g/kg wash 0.10 g/kg wash*
Product type anNBO, powder anNBO, liquid/gel
Heavy-duty laundry detergent, Colour-safe detergent 1.3 g/kg wash 0.70 g/kg wash
Low-duty laundry detergent 0.55 g/kg wash 0.30 g/kg wash
Stain remover (pre-treatment only) 0.10 g/kg wash 0.10 g/kg wash*
Table 1 Amount of aerobic non-biologically degrading substances (aNBO) and amount of anaerobe non-biologically degrading
substances (anNBO) for laundry detergents

Surfactants H400 Very toxic to aquatic life R 50

In concentrations < 25% in the product
Fragrances H412 Harmful to aquatic life with R52-53
Biocides used for preservation purposes long-lasting effects
Enzymes H334: May cause allergy or asthma R42
Bleach catalysts symptoms or breathing difficulties if inhaled
Enzymes H317: May cause allergic skin reaction R43
Bleach catalysts
NTA as an impurity in MGDA and GLDA H351: Suspected of causing cancer R40
Optical brighteners (only for heavy duty laundry detergent) H413 May cause long lasting effects
to aquatic life R53

Table 2 Exceptions for the general prohibition of substances endangering health or environment for dishwashing detergent

Surfactants H400 Very toxic to aquatic life R 50

In concentrations < 25% in the product
Fragrances H412 Harmful to aquatic life with R52-53
Biocides used for preservation purposes long-lasting effects
Enzymes H334: May cause allergy or asthma R42
symptoms or breathing difficulties if inhaled
Enzymes H317: May cause allergic skin reaction R43
NTA as an impurity in MGDA and GLDA H351: Suspected of causing cancer R40

Table 3 Exception from the general prohibition of health endangering substances or substances dangerous for the environment
of manual dish washing detergents

56 SOFW-Journal | 136 | 11-2010

 < Content

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WA
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Annex: List of affected classifications  Conclusion in Regard to the

Changes

GHS Hazard Statement EU Risk Phrase While there are no material changes in
regard to CDV-standards of laundry de-
H300 Fatal if swallowed R28 tergents and detergents for dishwashers,
H301 Toxic if swallowed R25 the standards for hand dishwashing de-
H304 May be fatal if swallowed and enters airways R65 tergents, all-purpose cleaners and sani-
tary cleaners have been considerably im-
H310 Fatal in contact with skin R27
proved. In view of the laundry detergents
H311 Toxic in contact with skin R24 and dishwashers the changed prohibi-
H330 Fatal if inhaled R23/26 tion of substances hazardous for health
and environment will have the highest
H331 Toxic if inhaled R23
impact.
H340 May cause genetic defects R46
H341 Suspected of causing genetic defects R68  Outlook
H350 May cause cancer R45
H350i May cause cancer by inhalation R49 The further development of award crite-
ria on basis of CDV in order to improve
H351 Suspected of causing cancer R40
environmental and health protection
H360F May damage fertility R60 will come to its technological limits in
H360D May damage the unborn child R61 the near future. A material reduction of
CDV will only be possible if the use of
H360FD May damage fertility. May damage the unborn child R60/61/60-61
colours and fragrances is reduced in the
H360Fd May damage fertility. Suspected of damaging the unborn child R60/63 foreseeable future. However, the new
H360Df May damage the unborn child. Suspected of damaging fertility R61/62 standards for sensitizing substances will
H361f Suspected of damaging fertility R62 probably result in innovations in the area
fragrances.
H361d Suspected of damaging the unborn child R63
H361fd May damage fertility. May damage the unborn child R62-63 Further information of the Federal Envi-
H362 May cause harm to breast fed children R64 ronment Agency under
H370 Causes damage to organs R39/23/24/25/26/27/28 http://www.umweltbundesamt.de/che
mikalien/waschmittel/zeichen.htm
H371 May cause damage to organs R68/20/21/22
and under the Internet presence of the
H372 Causes damage to organs R48/25/24/23
EU-Commission:
H373 May cause damage to organs R48/20/21/22
http://ec.europa.eu/environment/ecola
H400 Very toxic to aquatic life R50 bel/
H410 Very toxic to aquatic life with long-lasting effects R50-53
H411 Toxic to aquatic life with long-lasting effects R51-53
* Author’s address:
H412 Harmful to aquatic life with long-lasting effects R52-53 Marcus Gast
H413 May cause long-lasting effects to aquatic life R53 Umweltbundesamt
EUH059 Hazardous to the ozone layer R59 Fachgebiet IV 2.2
»Arzneimittel, Wasch- und
EUH029 Contact with water liberates toxic gas R29 Reinigungsmittel«
EUH031 Contact with acids liberates toxic gas R31 Wörlitzer Platz 1
EUH032 Contact with acids liberates very toxic gas R32 06844 Dessau
Germany
EUH070 Toxic by eye contact R39-41 Email: marcus.gast@uba.de
Sensitising substances

H334: May cause allergy or asthma symptoms or breathing R42
difficulties if inhaled
H317: May cause allergic skin reaction R43

58 SOFW-Journal | 136 | 11-2010

 < Content
S P EC I A LT I E S
M A R K ET R E P O RT

P. Morganti*, Yuan-Hong Li**

Wellness and Beauty Outside in:

China West – Working Together

To use Cosmetics and Functional Food is The research studies carried on the out- Traditional Chinese Medicine believes, in
a commonplace for consumers to sup- side and inside activity of cosme-nutra- fact, that the human body is an organic
port their Beauty and Wellness, partici- ceuticals have, thus underlined the con- entity where the appearances are re-
pating the body from an environment nection bridgeing the West and East flected from the inside.
every day more hostile to humans (1). medical culture (Fig. 2).
This the reason of the extraordinary in-
creasing of Cosmetics and Functional
Food on the West and East market dur-
ing the last 20 years (Fig. 1).
But innovative cosmetic and food prod- Global skincare market turnover ($ bn 2001 - 2010)
ucts have not only to increase the gen-
eral health of the body, but also strive to
stimulate the imagination and produce
emotions by combining exciting images,
sensual fragrances and tastes together
with a feeling of a caress on the skin and
mucous membranes, to convey the sen-
sation of total beauty effects.
Therefore, combination of effective Cos-
metics and Functional Food is the new
multiple approach to provide an excel-
lent skincare treatment that offers au-
dacity, shyness, pleasure and pride, to
obtain the total wellness and the con-
sumer desires.
The necessity to achieve this objective
has led to the NICE concept, in which the
Nervous, Immune, Cutaneous and En-
docrine systems work all together acti-
vating the skin physiology both from in-
side and outside, by the use of specific
Cosmeceuticals & Nutraceuticals, capa-
ble to stimulate the mind-body connec-
tion (2, 3).
Research studies, concentrated on the
skin activity of innovative Cosmetics and
Functional Food have shown that, im-
mune and cutaneous systems are strict-
ly connected to the nervous and en-
docrine systems, in such a way that these
four –way communication appear vital Fig. 1 Global skin care and nutraceutical markets
for our global wellbeing (4, 5).

SOFW-Journal | 136 | 11-2010 59

 < Content
S P EC I A LT I E S
M A R K ET R E P O RT

In fact, doctors of ancient China have al-

ways professed the strict relationship
between diet and health.
Thus, Sun Simiao from the IV century B.C.
wrote: »A good doctor first finds the
cause of illness and after finding it, tries
first of all to cure illness by food« (Fig. 3);
if the food doesn’t give results, then he
prescribes medicine because whatever
helps the body can be considered medi-
cine, and at the same time nourishment
(6).
For this reason each food in China was
and is until now considered important,
principally for its therapeutic properties.
Therefore Nei Jing (2698-2598 b.C.) wrote:
»Take five cereals as base nutrients, five Fig. 2 The cosmetic bridgeing between East and West culture is in progress
fruits in addition«.
The five meats to invigorate, and the five
vegetables as supplements, then, mix to-
gether the Qi and the Wei (flavors) of the
diet, this mixture is what benefits the
mind and body (7)
However, meanwhile the Official West
Medicine is anchored on the pharmaco-
logical efficacy of single ingredients, ac-
tive on single human organs, or tissues,
people worldwide know how the skin is
a mirror of our soul and how emotions
can influence illness.
And this was the adage of antique Ro-
mans also: mens sana in corpore sano (an
healthy mind in an healthy body).
Thus staying healthy is the general re-
quest of women and men until now.
Being beautiful is no more enough, peo-
Fig. 3 Sun Simiao (IV century B.C.) wrote that illness at the beginning has to be
ple want to feel beautiful and healthy in
cured by food
their body and in their mind too.
But what’s the real skin-body connec-
tion?
The skin is equipped with an effective
communication and control system, de-
signed to protect the human organism
in a constantly changing environment
(Fig. 4).
Thus recently, biologists determined that
hormones of the endocrine system help
the immune and nervous systems defend
the body; stress hormones, for example,
can initiate actions in the brain and im-
mune system, in response to injury or mi-
croorganisms, to act as an immune sys-
tem regulator (8, 9).
On one side, the immune molecules cy-
tokines can initiate brain actions, trig-
gering feelings or sluggishness, sleepi- Fig. 4 The skin is equipped with a complex communication control system
ness and loss of appetite.

60 SOFW-Journal | 136 | 11-2010

 < Content

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 < Content
S P EC I A LT I E S
M A R K ET R E P O RT

On the other hand, the skin is equipped

with a dense network of highly special-
ized efferent sensory and efferent auto-
nomic nerve branches, designed to pro-
tect all the human organs, in a con-
stantly changing environment, by an ef-
fective communication and control
system regulated by the activation of cy-
tokines, chemokins etc.
In fact, the sensory system contains re-
ceptors for touch, temperature, pain, itch
and other physical and chemical stimuli.
The information is either processed in
the Central Nervous System as well as
may directly elicit an inflammatory re-
action by antidromic propagation of
these impulses.
The effector function of a nerve may be
Fig. 5 The overlap among environment, life style nutraceuticals beauty foods cos-
determined by secreted neuropeptides
meceuticals, cosmetics and medical devices
and the corresponding receptors of tar-
get structures (10-14).
In addition, these neuropeptides seem to
exert multiple effects on immunecom-
petent cells thus, suggesting a strong in-
teraction between the nervous and the
immune systems.
In contrast to sensory nerve fibers, the
distribution of autonomic nerves is re-
stricted to the dermis,innervating blood
vessels, arteriovenous anastomoses, lym-
phatic vessels, glands, hair follicles, and
stimulating immune cells to release neu-
rotrasmitters, such as acetylcholine and
catecholamines.
However, the overlap among environ-
ment and life style and the outside and
inside activity of Cosmetics products and
Functional Food are fundamental to
regulate the role of keratinocytes and
Fig. 6 Specialized signals alert both the immune and neervous systems
fibroblasts, capable to orchestrating the
complex sensory and immunological
events (Fig. 5).
These specialized cells, communicate each
other by the production of specific sig-
nals, the cytokines, speaking with the
stratum corneum, meanwhile special pep-
tides are produced to communicate with
dermis.
Thus, specialized peptides alert the
immune systems as well as specific
hormones activate the nervous fibers
(Fig. 6).
Therefore, this orchestra of signals have
different ways of communication among
cells, tissues and organs by a botton-up
Fig. 7 Bi-directional communication systems between human cell and body:
(from dermis ↑) or top-down (epidermis
bottom-up & top-down
↓) direction (15, 16) (Fig. 7).

62 SOFW-Journal | 136 | 11-2010

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S P EC I A LT I E S
M A R K ET R E P O RT

try to protect the skin from endogenous

and exogenous stresses, while preserving
its healthy and young look for a longer
period of time.
Thus, the incoming age of biotech-Cos-
metic and Beauty Food, together with
the development of nano-science and
dermogenetics, represents the future
challenge to solve the new consumers
needs of an evolving Society.
Moreover, it would be nice the today
scientific/collaboration between Italy
and China will again become an eco-
nomical continuing collaboration as al-
ready happened during the period of:
Marco Polo (1271–1292 Chinese period)
and Matteo Ricci (1582–1610 Chinese
period) (Fig. 10).
Fig. 8 Environment and irregular life-style are the main outlines of the premature
skin ageing

All these signals are necessary, in fact, for

the cell living, modulating all its nutri-
tive and survival necessities connecting
the skin with the gland system also,
through the production of hormones such
as, catecholamines, oestrogenic and an-
drogenic compounds.
However, the environment’ aggression
and the skin aging process create an ev-
ident imbalance of all these molecular
signals that go hand in hand with the cell
renewal (Fig. 8).
Thus Synthesis of hyaluronic acid is re-
duced, production of elastin and colla-
gen is altered and premature aging be- Fig. 9 The first mind-body skincare
come visible.
Therefore, the necessity of »intelligent«
Cosmetics and Beauty food, capable to Food, reactivating the skin dynamic equi-
activate and promote the different in- librium, are modifying the Cosmetic and
tercellular signals conveyed at level of diet Supplement Market.
the skin, contributing to the epidermal These innovative products, having active
barrier function (16). and no more passive mechanism of ac-
For all these reasons the new generation tion, appear more efficacious than nor-
of cosme-nutra-ceuticals based on the mal cosmetic and food, both in the short
NICE approach represent not only the and in the long period of time (18,19).
bridge between the skin and our image The consumer demands for more active
recorded from our brain, but also the re- and safety Cosmeceuticals & Nutraceu-
al connection between the West way of ticals and the emerging technologies cre-
thinking and living and the Chinese ate every day innovative skin-care beau-
holistic culture. ty products to be used both from inside
Because of this skin-mind connection, and outside.
the first mind-body skincare are appear- This is why the existing direct relation-
Fig. 10 Marco Polo and Matteo Ricci
ing on the market (17) (Fig. 9). ship between Cosmetic and Beauty Food,
two Italians connected with the Chi-
What is interesting to underline is that establishing more knowledge, new ana-
nese culture
NICE- Cosmetics and NICE-Functional lytical tools and therapeutic modalities,

SOFW-Journal | 136 | 11-2010 63


S P EC I A LT I E S
M A R K ET R E P O RT
The topic reported on this paper will be again dis-
cussed in occasion of the next 10th ISCD Congress
»Life Science Meet Cosmetology« that will be held
in Shenyang (China) May 30-31 June 1, 2011).
References
(1) Morganti P. (2009) Beauty from the inside
and the outside. Natural products work in
multiple ways. In: Nutritional Cosmetics.
Beauty from within . A. Tabor and R. Blair eds.
William Andrew pub., Elsevier Inc, UK pp 93-
111
(2) O’Sullivan RL, Lipper G, Lerner EA. (1998). The
Neuro-Immuno- Cutaneous-Endocrine Net-
work: Relationship of mind and skin. Arch.
Dermatol 134: 1431-5
(3) Hosoi J. (2009) Establishment and change of
NICE approach. J.Appl. Cosmetol. 27: 200
(4) Hosoi J, Murphy GF, Egan CL, Lerner EA, Grabbe
S, Asahina A, and Granstein RD (1993). Regu-
lation of Langerhans function by nerves con-
taining calciotomin gene-related peptide.
Nature 363: 159-163
(5) Torii H, Yan Z, Hosoi J, and Granstein RD (1997).
Expression of neurotrophic factors and neu-
ropeptides receptors by Langerhans cells and
Langerhans cell-like line XS52: further sup-
port for a functional relationship between
Langerhans cells and epidermal nerves. J. In-
vest. Dermatol. 109: 588-599
64
< Content
(6) Sun Simiao (400 B.C.) Ben - Cao (Antique Chi-
nese Pharmacopeia)
(7) Zhang Daoling (A.D. 25- 220) Zhengyi Meng
Wei (Maighty Commonwealth of the ortho-
dox oneniss)
(8) Lintner K and Peschard O (2000) Biologically
active peptides: from a laboratory bench cu-
riosity to a functional skin care product. Int.
J. Cosm. Sci. 22: 207-218
(9) Roosterman D, George T, Schneider SW, Bun-
nett NW, Steinhoft M (2006) Neuronal con-
trol of skin function: the skin as a neuroim-
munoendocrine organ. Physiol. Rev. 86 (4):
1309-1379
(10) Schauer E (1994) Propiomelacortin-derived
peptides are synthetised and released by hu-
man keratinocytes. J. Clin. Inv. 93: 2258-64
(11) Scholzen T Armstrong CA, Bunnett NW, et al
(1998) neuropeptides in the skin: interactions
between the neuroendocrine and the skin im-
mune system. Exp. Dermatol 7:81-96
(12) Schmelz M, Michael K, Weidner C et al. (2000)
Which nerve fibres medicale the axon reflex
flare in human skin? Neuroreport 11: 645-648
(13) Metze D, Luger T (2001) Nervous system in the
skin. In: Frenkel RK, Woodley D (Eds). The bi-
ology of the skin N.Y, Taylor & Francis, pp 153-
176
(14) Ikoma A, Steinhoff M, Stander S et al. (2006)
The neurobiology of itch. Nat. Rev. Neuro Sci
7: 535-547
(15) Elias PM (1983) Epidermal lipids, barrier
function and desquamation. J. Invest. Der-
matol. 80 (suppl.):44s- 49s
(16) Sudhesh J (2000) Investigation into the ef-
fects of music and meditation on galvanic
skin response ITBM-RTBM21 158-162
(17) Morganti P (2009) Dal controllo delle emo-
zioni…..la bellezza globale. Farmacia News
n°10: 64
(18) Morganti P, Morganti G, Fabrizi F, Cardillo A
(2008) A news sun to rejuvenate the skin. J.
Appl. Cosmetol. 26: 159-168
(19) Morganti P, Palombo M, Palombo P, Fabrizi G,
Cardillo A, Carezzi F, Morganti G, Ruocco E
and Dzierzgowski S (2010) Cosmetic Science
in Skin Aging: Achieving the Efficacy by the
Chitin Nano-Strucured Chrystallites SOFW
Journal 136: (n3): 14-24.
Authors’ address:
Pierfrancesco Morganti*
Professor of Applied Cosmetic
Dermatology
II University of Naples
Visiting Professor of China Medical
University Shenyang
Head of R&D
Mavi Sud s.r.l
Rome, Italy
I.S.C.D President and Secretary General,
Rome, Italy
** Yuan-Hong Li
Department of Dermatology
No. 1 Hospital of China Medical
University of Shenyang
China
SOFW-Journal | 136 | 11-2010
 < Content

Cosmetology – Theory and Practice

Research, Test Methods, Analysis, Formulas Now o
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The aim of creating this book was to provide cosmetic chemists, dermatologists, pharmacists and
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2 Test Methods –
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homeostasis and tissue regeneration is
66
< Content
diminished, eventually resulting in in-
trinsic skin ageing. Stem cell ageing is
further accelerated by extrinsic factors
such as air pollution, daily sunlight, and
ozone. Thus, damage of stem cells is a
predominant factor that leads to accel-
erated skin ageing.
RAHN has screened two cosmetic ac-
tives, DEFENSIL® and CELLIGENT®, for
their stem cell protecting properties and
have observed impressive results for
both.
DEFENSIL® is an optimised system for
physical and biochemical protection
which normalises sensitive skin types
due to its excellent inflammation in-
hibitory efficacy.
Treatment with DEFENSIL® leads to re-
markable effects on stem cells following
damage by UV-irradiation:
– Enhancement of colony forming effi-
ciency of epidermal stem and prog-
enitor cells upon cell stress (in vitro
study)
– Protection of stem cells and their
ability to form a 3D epidermis upon
cell stress (in vitro study)
DEFENSIL® provides resistance to the key
cells involved with skin homeostasis and
thus counteracts the appearance of irri-
tated and stressed skin.
CELLIGENT® is both a preventative and
an active cell and DNA protector. The ac-
tive ingredient complex has been spe-
cially developed for treating UV-stressed
skin.
CELLIGENT® shows excellent stem cell
protecting properties following expo-
sure to UV-stress:
– Maintenance of stem cell tissue
restoration capacity upon UV-stress
(in vitro study)
CELLIGENT® maintains the function of
epidermal stem cells, is an intelligent
stem cell and DNA protector, and there-
fore should form part of any modern an-
ti-ageing product (cosmetics@rahn-group.
com).
■
A New Ecological Range of
Cleaning Products
Doronwell Limited is launching a new
ecological range of cleaning products for
the European Cosmetic Industry.
Doronwell has already been providing
a cost effective approach to cleaning
problems in the European pharmaceuti-
cal and other industries for many years.
As Dober’s agent for the Chematic range
we have detergents that can clean prod-
ucts such as waxes, TiO2 and other hard
to clean ingredients.
Doronwell and Ecover have worked to-
gether on creating a range of effective,
efficient and ecological cleaning prod-
ucts suitable for use in cosmetics pro-
duction facilities. The result is DNW ECO-
GOLD, DNW ECO-PURE and DNW ECO-
FORCE. DNW ECO-PURE is formulated us-
ing Ecover’s new Eco -Surfactants and
can be used as a cleaning agent in Eco-
cert certified cosmetics production facili-
ties (www.doronwell.com, Email: ken@
doronwell.com).
■
Kelisoft™ –
A Hair Growth Minimiser for
Beards and Underarms
Kelisoft™ slows down hair growth while
refining its diameter. The hair is thus less
visible and the need to shave as often is
reduced. Thanks to its anti-inflammato-
SOFW-Journal | 136 | 11-2010

ry properties, Kelisoft™ also provides an
immediate soothing sensation after hair
removal. Kelisoft™ contains a highly pu-
rified natural plant molecule, which regu-
lates the production of inflammatory
mediators (PGE2, IL8, VEGF) known to
stimulate hair growth and which are in-
volved in the burning feeling after shav-
ing.
Twice daily application of a roll-on con-
taining 3% Kelisoft™ under the arms
of 20 female volunteers for 2 months
demonstrated that hair grows slower
before
after
and finer, so women can skip a day be-
tween shaves. Additional tests carried
out on the thick dark beard of male vol-
unteers confirmed that once daily appli-
cation of a gel-cream containing 4%
Kelisoft™ for 2 months reduces the hair
growth speed (»5 o’clock shadow« de-
layed by 3 hours) and provides a sooth-
ing sensation after shaving.
Kelisoft™ has been specifically devel-
oped for deodorant applications, 2 ver-
sions are available: a water soluble one
and Kelisoft™OS which is oil soluble.
Kelisoft™ is the right ingredient for for-
mulations aimed at reducing the fre-
quency of shaving while keeping the skin
soft and smooth (Sederma SAS 29 rue du
Chemin Vert 78612 Le Perray en Yvelines
cedex France, Tel: ++33 1 34 84 10 10,
Email: sederma@sederma.fr).
■ for skin care as well as hair and body
SOFW-Journal | 136 | 11-2010
< Content
40 Cognis Products Certified
with NaTrue Label
Since March 2010, NaTrue (a label from
the International Natural and Organic
Cosmetics Association) offers the possi-
bility to apply for raw material certifica-
tion. Cognis is one of the first suppliers
of functional cosmetic ingredients to gain
NaTrue certifi-
cation for a
broad range of
its raw materi-
als. In order to
serve the green
trend, Care
Chemicals’
customers can
use these ingredients in NaTrue-certified
natural and organic cosmetics as well as
natural cosmetics with organic portions.
This makes it much easier for them to
formulate end products that should
comply with the NaTrue guidelines.
An independent certifier has confirmed
that the Cognis products applied for
(cosmetic oils, emulsifiers, surfactants,
etc.) fulfill the criteria of being natural
or naturally derived, including detailed
investigations on every step of the pro-
duction process plus an audit. This pro-
vides manufacturers aiming to formu-
late and market natural cosmetics clear
advantages.
The main NaTrue label criteria according
to the definition of the International
Natural and Organic Cosmetics Associa-
tion are:
• Only water, natural ingredients, de-
rived natural ingredients and nature-
identical ingredients are permitted.
• Manufacturing processes for Natural
Cosmetics and their raw materials are
strictly regulated.
• Strict minimum for natural sub-
stances and maximum levels for de-
rived natural and organic substances
apply for all three NaTrue cosmetic
classification grades depending on
the application purpose: Natural Cos-
metics, Natural Cosmetics with Or-
ganic portion and Organic Cosmetics
Cognis has created special formulations
R AW M AT E R I A L S
cleansing that comply with NaTrue’s
guidelines for the three categories.
The 40 certified Cognis products include
surfactants, emulsifiers, consistency wax-
es, emollients, cream bases, protein hy-
drolysates as well as a pearlshine wax
dispersion and a hair conditioning boost-
er. All ingredients are listed on the Na-
True website: http://www.natrue-label.
com.
■
To Regulate the Level of
Tolerance and the Long-Term
Balance of the Skin –
Soothing
Unisooth ST-32 stimulates the skin de-
fense by boosting the immunity mecha-
nism of the skin and by protecting the
Langerhans cells
Unisooth PN-47 modulates the skin re-
activity by using powerful JAK/STAT in-
hibitors
Unisooth EG-28 modulates the skin re-
activity by using powerful NF-kB in-
hibitors
Unisooth PN-47 and EG-28 combined,
they block the keratinocyte activation
loop.
Induchem AG is represented in India by
N V Organics Pvt. Ltd. You are welcome
to contact them at booth 212, HPCI Con-
gress, India.
■
Scientific Data on Skin
Penetration Behavior of
PARSOL® SLX
PARSOL® SLX is the first polymeric sun-
screen that got approval by the EU and
other major authorities around the world
including Japan, China and Australia. The
polymeric product has an average mole-
cular weight of more than 6 000 and
consists of a polysilicone backbone with
UVB chromophores covalently attached
to it.
In the scientific study, the behavior of ra-
dio-labeled material of PARSOL®SLX has
been investigated in a standard design in
an in vitro skin penetration study with
donated human skin samples exposed
67
 < Content
R AW M AT E R I A L S / FO R MU L AT I O N

over a 24 hours period to a sunscreen
product containing 10% PARSOL® SLX.
The result demonstrates that more than
99.5% of the applied amount of PARSOL®
SLX can be retrieved after a 24 hours ex-
posure period from the skin surface and
the stratum corneum. This result sup-
ports the concept that polymeric sun-
screen actives can be designed and, con-
sistent with other polymeric substances,
will show a very low, if any, skin pene-
tration rate.
PARSOL® SLX’s overall good safety pro-
file combined with the argument of the
very low or no skin penetration rate al-
lows sunscreen manufacturers to design
sunscreen products for sensitive skin, in-
cluding products for babies and children
with a favourable skin feel (DSM Nutri-
tional Products, www.dsm.com).
■
DURAGREEN™ 135 Polymer –
First Zinc-free Technology for
Household Floor Care Market
As part of its strategy to expand its port-
folio of sustainable solutions to meet the
evolving needs of the global floor care
industry, Dow Fabric & Surface Care to-
day the introduction of DURAGREEN™
135 Polymer, a new zinc-free film former
for household floor care applications.
DURAGREEN™ 135 Polymer provides a
number of environmental advantages
and performance benefits to formula-
tors of floor care products. It enables the
formulation of high-gloss polishes with-
out the use of undesirable ingredients,
such as tributoxyethylphosphate, fluo-
rosurfactants, and silicone defoamers,
thereby enabling formulators to opti-
mize their products by reducing the
number of ingredients used. Formula-
tions based on DURAGREENTM 135 Poly-
mer are suitable for use on PVC
(polyvinyl chloride) and linoleum floors,
and provide a number of key benefits to
the consumer, including:
• Good wear resistance
• Excellent detergent resistance
• Excellent removability
Dow Fabric & Surface Care, www. science
ofcare.com.
■

FO R MU L AT I O N

LIGHTEN UP – Skin Color Refining Wipes – Light, fast absorption, whitening

Component INCI % Function
COSMEDIA® ULTRAGEL 300 Polyquaternium-37 0.5 Polymer, cationic
Water, demin. Water 91.5
EUMULGIN® VL 75 Lauryl Glucoside (and) Polyglyceryl-2 Dipolyhydroxy- 1.0 Emulsifier (O/W
stearate (and) Glycerin
ACTIWHITE® LS 9808 provisional Water (and) Glycerin (and) Sucrose Dilaurate (and) 2.0 Active ingredient
Polysorbate 20 (and) Pisum Sativum (Pea) Extract
Perfume, preservative Perfume, preservative q.s.
Ethanol (denatured) Ethanol 5.0

Preparations in the laboratory:

Cold process.
Disperse Cosmedia® Ultragel 300 in water while stirring to make a homogeneous gel.
Add all other ingredients one after another to the gel while stirring until a homogeneous lotion is formed.
Adjust the pH-value if necessary and add Ethanol at the end

Quelle: Cognis GmbH, Rheinpromenade 1, 40789 Monheim, www.cognis.com

68 SOFW-Journal | 136 | 11-2010

 < Content


 
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EVENT
SOFW
Media Info 2011
Advertisement prices, editorial &
ad deadlines, events calendar,
circulation analyses, information
on all language editions.
All the information you need
at one glance.
Advertising Contact:
Siegfried Fischer
E-mail: s.fischer@sofw.com
Tel.: +49 821 32583-16
Fax: +49 821 32583-23
Download our media info at
www.sofw.com
(→ SOFW-Journal → Mediadata)
www.sofw.com
70
< Content
PCHi Opens for
Pre-Registration
22–24 February 2011
Shenzhen Convention & Exhibition
Centre
The pre-registration for PCHi is now
open. Visitors and buyers are encouraged
to pre-register at the event website
(http://www.pchi-china.com/en).
The current pre-registration site presents
a more user-friendly interface, an easier
login function, and a more streamlined
registration process. Visitors may also in-
vite their colleague to pre-register by
simply submitting their names and email
addresses.
A number of benefits follow pre-regis-
tration. The iCatch platform is available
to pre-registered visitors. The platform
provides information on exhibiting com-
panies, such as company profiles, new
product information, and press releases.
On-site meetings can also be planned
months before the show using iCatch.
Another benefit upon pre-registration is
that visitors will receive regular updates
about PCHi through pre-event e-news-
letters. These provide information to
keep buyers well-informed of industry
news and PCHi happenings, in the spirit
of enhancing and extending the PCHi
experience for all participants.
At present, information about the annu-
al PCHi Conference Programme has also
been announced. There are five themes
to be presented at PCHi 2011 by indus-
try-leading experts: Technology, Formu-
lators’ Workshop, Regulatory Summit,
Marketing Trends, and New Technology
Session & New Technology Feature Areas.
In keeping with the industry’s needs, the
Technology conference comprises three
sub-themes: New Technology & Innova-
tions, Earth-Friendly Formulations, and
The Macho Side of Personal Care.
Reed Sinopharm Exhibitions is targeting
6 000 pre-registered PCHi visitors before
the site closes nearer the event date.
Those planning to attend PCHi 2011 have
until 15 February 2011 to pre-register to
avoid entry fees of 100 RMB and wait-
ing lines at the event venue.
Rousselot Chooses PCHi to
Reach Chinese Customers
Rousselot (China), a gelatin & hydrolyzed
collagen company, had the good fortune
of working with Reed Sinopharm Exhibi-
tions (RSE) at API China, an exhibition for
the pharmaceutical manufacturing sec-
tor, before it had to make a trade show
marketing decision for its personal care
product line of hydrolyzed collagen this
year. The professionalism of the RSE
team at API left a strong and lasting im-
pression on Rousselot. So when the com-
pany was looking to participate in its
first trade show for the cosmetics man-
ufacturing sector in China, API’s sister
show Personal Care and Homecare In-
gredients (PCHi) was a natural choice.
Rousselot will be exhibiting for the first
time at PCHi 2011.
At PCHi 2011, Rousselot will be show-
casing Peptan, its unique line of hy-
drolyzed collagen, used for skin care and
healthcare purposes. While widely known
as a beauty product, collagen also im-
proves bone and joint health. It is main-
ly consumed orally or by application. The
company prides itself in producing the
»perfect« collagen. The taste, odour,
colour and molecular weight of the
product are important factors to consid-
er, and Rousselot always strives to
achieve excellence in each of these four
attributes.
Rousselot is part of the VION group of in-
gredient manufacturing companies. The
company’s two main products are gelatin
for the pharmaceutical and food indus-
try, and hydrolyzed collagen for the nu-
tricosmetics and bone health markets. As
the world’s largest producer of gelatin
and collagen, Rousselot has 12 factories
worldwide, including one in France man-
ufacturing its Peptan™ brand of hy-
drolyzed collagen. The company entered
China in 1996 offering gelatin to the lo-
cal market, and started marketing hy-
drolyzed collagen ten years later, in
2006.
■
SOFW-Journal | 136 | 11-2010
 < Content

Global Ingredients &

Formulations Guide 2009
Innovations – Challenges and Chances

GLOBAL INGREDIENTS & • REACh • economic solutions • new ingredients

FORMULATIONS GUIDE 2009 • new trends • new technologies
• new formulation techniques • formulations
Innovations – Challenges and Chances
• company index • suppliers
In critical times it is even more important to consider innovations.
Only innovative companies can master the crisis and keep their po-
sition in the global market place.
A German proverb says: ”Necessity is the mother of invention”. This
saying gains new meaning these days. At all times humans had to
face crisis and overcame it with new ideas.
Today the motto must be: ”Innovation masters crisis”. This is also
true for cosmetic and personal care products. To neglect research and
development in these days for cost saving reasons, can be fatal in
the future. Innovative competitors can gain ground and strengthen
their market position. Therefore for all producers of cosmetic and
personal care products it is essential to know all innovations in for-
mulation techniques and ingredients.
Verlag für chemische Industrie
H. Ziolkowsky GmbH, Augsburg
The GI&FG 2009 focusses on this subject (see content overleaf).

• 532 Pages • 10th Edition • German Version available also

ORDER FORM Order-No. 500-1059

❍ ____ Copy(ies) Global Ingredients & Formulations Guide 2009 · Innovations – Challenges and Chances ISBN: 978-3-87846-272-9
Price: € 75.– / sFr 130.– (+ Shipping and Handling)

Company Customer-No.

Name Tel.

Street / P.O. Box Fax

Zip Code, City E-mail

VAT Id. No. (if EU) Date, Signature

PAYMENT
Send your order to:
❍ Credit card ❍ Visa ❍ Amex ❍ Mastercard
❍ Invoice/ Bank Transfer · Verlag für chemische Industrie, H. Ziolkowsky GmbH
· P.O. Box 10 25 65 · 86015 Augsburg · Germany
Name of Card Holder · Tel: +49 (0) 8 21 3 25 83-18 (Olaf Rossmann)
· Fax: +49 (0) 8 21 3 25 83-23 · E-Mail: rossmann@sofw.com
Card-No.

Security Code Exp. Date Order online: www.sofw.com/books

 < Content

Global Ingredients & Formulations Guide 2009 – Content

Innovation and REACh Innovation and New Trends

REACh in Practice, 5th International Fresenius Chemicals Matrix Proteins of the Papillary Dermis – Primary Targets
Policy Conference (C.S. Koch) of Intrinsic Dermal Aging? (D. Boudier, V. Dubreuil,
L. Marchand, N. Guichard, B. Closs)

Innovation and Economic Solutions A New Generation of Green Surfactants and Emulsifiers

Feasibility Studies Regarding Manufacturing of Submi- Solving Low pH Formulating Challenges with Naturally
cron Systems Using a New ”Rotor/Rotor”-Processing Derived Thickeners (D. Peterson, M. Tolchinsky)
facility (G. Kutz, V. Salzmann, M. Weber, A.
Lewandowski)
Innovation and New Technologies
3D In Vitro Technology: Modelled Skin Helps Animals in
Innovation and New Ingredients Reaching Relief (C. Rüffer)
Luvigel® Advanced (Polyquaternium-86) – A New Cat- Molecular Biological Mechanisms of Skin Aging
ionic Thickener with Styling and Conditioning Benefits (W. Voss, G. Schlippe)
(P. Hoessel, J. Basilan, S. Nguyen-Kim)
Beneficial Fatty Oils for Promoting Healthy Skin
Novel Graft Polymer Boosts SPF Performance
(L. Marlier, J. Cincotta, T. Martin, W. Van Drunen) A New Strategy in Anti-Aging Cosmetics: DORMINS –
Give your skin and hair a rest (V. Maienschein)
Delivering Softening Benefits from a Shampoo System
(T. Gao, J.-M. Tien, A. Bidaye, S. Cardinali, and J. Kinney)
SKINASENSYL® – The Innovation from Neurocosmetics Innovation and Formulation Techniques
Research for Calming Hypersensitive Skin HYBRID – Photostable UVA Protection; For sun protecti-
(J. Ivosevic-Zaper) on and daily care products
A Patented Multifunctional Ester for Personal Care A Formulation Strategy for High SPF Sun Care Products
Applications (J. Hewitt)
(L. Bouldin, D. Smith, M.K. Smith, R.J. Smith)
OLEOSOMES – Natural Emulsifying Systems Formulations
(D.C. Long, J. Ferguson)
• Disclaimers for the formulations • Skin Care • Hair
Cococin™ – The Nourishment Factor®: A Natural Reser- Care • Body Care • Decorative Cosmetic
voir of Nutrients for Beauty from the Inside and Out
(L. Prakash and M. Majeed)
Polyether-Modified Silicones for Cosmetic Applications Company Index
(J. Newton, I. Vervier, B. Durand, and S. Masse) Contact addresses, company description, product
Interleukin-1 Alpha – An Epidermal Cytokine Critical for range, service
Skin Renewal (I. Pomytkin)
MIRAGE Borosilicate Pigments – Highlights for Subject Index
Decorative Cosmetics (K. Steinbach, U. Schmidt)
Dermofeel® sensolv: A Skin-Feel Like Silicon – But
Without Silicon
Suppliers’ Directory
Symbio®muls GC: The New Natural ”One-Fits-All-W/O-
Emulsifier”
Dermofeel® P-30: Natural Hair Care – A Paradoxon?

www.sofw.com

Azelis Appoints Benoit Fritz
as Principals Development
Director
Benoit Fritz is highly experienced, hav-
ing spent the last 8 years at The Dow
Chemical Company as Commercial Di-
rector for Dow Wire & Cable business in
EMEA and previously, as Market Manag-
er for the same division. He has also
worked with DuPont-Dow Elastomers as
the global Wire & Cable team leader for
ethylene copolymers and as European
sales development manager for DuPont-
Dow ethylene copolymers. The appoint-
ment of Benoit Fritz continues to build
on Azelis’ structured principal develop-
ment, a cornerstone in Azelis’ business
progress. This will enable Azelis to create
long term sustainable growth and value,
combined with sensible customer focus.
Benoit holds a Master’s Degree in Or-
ganic and Bio-organic Chemistry at the
Pierre & Marie Curie University in Paris.
In June 1997, he conducted a DEA re-
search at Ecole Normale Superieure
Paris.
■
Formulae and Dossiers
Online on Internet
Something new is available for IT ser-
vices to the Personal Care Industry in-
troduced by Logos International, a com-
pany committed to IT Cosmetic Industry
since 1990 on the international market.
The new solution is an innovative service
online to the cosmetic companies based
on the Internet
Data and software are hosted by a
provider who grants the continuous ac-
cess and the due protection on data
against unwanted access and for data
backup copies. The service is web based
thus the user has just to enter the url ad-
dress to run immediately R&D, Product
Dossiers and Quality Assurance & Con-
trol.
With this new service frees Personal Care
companies from many worries and costs:
• No need of server computer.
• Stop taking care of data backup
copies.
SOFW-Journal | 136 | 11-2010
< Content
• No need to purchase basic software
licenses, like MS SQL, etc.
• Stop depending on costly and often
little available specialized IT people
for LAN settings, permissions,
authentications, etc.
This new service increases the flexibility
and decreases costs: It is no more neces-
sary to depend on certain fixed comput-
ers. Formulators can access their dataset
–formulae, ingredients, regulatory, sta-
bility, etc.- anytime from any computer
inside and outside the company. It is
even possible the use of smart phones.
The highest level of protection, reliabili-
ty and secrecy on data is assured. The
server offered by Logos is protected with
the best and most powerful firewall sys-
tems. There no strongbox safer than the
one provided by Logos. Even the net
transactions are encrypted.
The communication speed is the one that
all of us already know, but it is destined
to be increased for the continuous in-
vestments that the entire world does al-
most daily to increase the quality and the
speed.
The low-cost philosophy has been adopt-
ed: the entry level is 80 euro all-inclusive
per month per active workstation (+ ac-
tivation cost). A cost so low must not de-
ceive and must not make thinking to a
limited or elementary system. 4MACS is
a vast, ultra-complete and ultra-reliable
system, with multi-language and multi-
media features, often exceeding the best
expectations, matured in over 20 years
of multi-national experience. The system
is very helpful to small and medium com-
panies but mostly to large companies,
those companies having R&D labs scat-
tered in different locations and often in
different Countries. The potentiality of
4MACS is enormous: virtually thousands
of users can be active simultaneously
and the database is capable of hundreds
of thousands of formulae.
Logos International assures a constant
system updating to satisfy the current
and future European requirements as ex-
pected in the near future regarding the
product notification, toxicology, REACh
requirements, etc. Data exporting and
data interchange with other systems are
included.
COM PA N Y N E W S
The activation of the service for a new
customer is virtually immediate since
server and software are already avail-
able. The procedure is »get the creden-
tials and run the system«.
Logos Int’l assures to customers all relat-
ed services like training sessions, tele-
phone, and online assistance. Logos Int’l
is also available to satisfy possible par-
ticular custom necessities.
Manufacturing Personal Care Companies
may now look at new operative scenar-
ios having finally the possibility to use of
the latest Internet technology to run
formulae, ingredients, QA&QC and prod-
uct dossier with the following remark-
able advantages:
• Company structure more streamlined,
flexible and versatile: no server and
no server attendant needed.
• Company formulae, ingredients and
product dossier data accessible from
any computer
• Usage of managerial specialized soft-
ware extremely complete and effi-
cient and always up-to-date
• Hardware and software costs defi-
nitely cheaper than before
• Usage of the latest IT technology
The online service can be installed on de-
mand on customer’s hardware system
with the same modality and same costs.
The license of use purchase is also avail-
able (Logos International, Via Angelo
May 16, 24121 Bergamo Italy, info@
logwin.com, www.logwin.com).
■
Gattefossé Announces New
GMP Certification for its
Cosmetics Operations
Gattefossé is proud to announce that it
is one of the first manufacturers to re-
ceive a certification of compliance with
the GMP guidelines published in 2008 by
the European Federation for Cosmetic
Ingredients (EFfCI).
The quality of cosmetic ingredients is
critical to ensure their safety and effica-
cy. For Gattefossé, the application of ap-
propriate good manufacturing practice
(GMP) principles to cosmetic ingredients
73

COM PA N Y N E W S
is essential to ensure the supply of high
quality ingredients.
Gattefossé’s main industrial plant locat-
ed near Lyon, France was audited and
certified to be in compliance with the
GMPs for Cosmetic Ingredients, by SGS,
an international independent certifying
body.
The GMPs for Cosmetic Ingredients have
been established by EFfCI (European Fed-
eration for Cosmetic Ingredients), GMP
Guide (2008), and are based on the ISO
9001 structure and the IPEC (Interna-
tional Pharmaceutical Excipients Coun-
cil) GMP guide as a reference.
Good Manufacturing Practices (GMP)
Guidelines cover all aspects of the man-
ufacturing of cosmetic ingredients from
starting material to finished products.
This is the second time Gattefossé has
voluntarily sought GMP compliance cer-
tification, in 2008, Gattefossé was one of
the first pharmaceutical excipient man-
ufacturers to be inspected by the French
Healthcare Authority (AFFSAPS) and ob-
tain certification of compliance to GMP
for raw materials for pharmaceutical
use.
■ expertise includes formulation, product
74
< Content
F I R M E N N AC H R I C H T E N
Shastry Appointed
Technology Manager for
Arizona Chemical
Arizona Chemical announces that Ra-
machandra Shastry, Ph.D. has been ap-
pointed Technology Manager for the
Specialty Products Group. His responsi-
bilities include lead-
ing the technolo-
gy assessment
and develop-
ment of Spe-
cialty Poly-
meric Gel-
lants for
personal care
applications and
providing consulta-
tive solutions to support customer for-
mulation development. Dr. Shastry joins
Arizona Chemical following successful
tenures as the Director of R&D at Accu-
pac, and Medicia. His prior experiences
include positions as Manager at the Nat-
ural Dentist and as Senior Scientist at
Gillette and Colgate Palmolive Co. His
and process development in skin care,
hair care and oral care applications. Dr.
Shastry received his doctorate in Mate-
rials Science/Solid State Chemistry from
the Indian Institute of Science, Banga-
lore, India.
■
Aqua Bio Technology:
Sales Agreement
Aqua Bio Technology, a Norwegian ma-
rine biotech firm developing, producing
and selling patented marine based in-
gredients and technologies to the inter-
national cosmetic and skin care industry,
today announced it has entered into a
sales agreement with a highly regarded
Asian manufacturer of skin care prod-
ucts.
As a part of the agreement, ABT will sup-
ply its bioactive ingredient Zonase X (™)
to be the key ingredient in a new prod-
uct line the manufacturer will launch in
January 2011. The new line will consist of
five products, developed in collaboration
with ABT.
The new skin care line will initially be in-
troduced in the Asian markets through
the manufacturer's existing distribution
channels. In addition, the manufacturer
is also in discussion with distributors in
Australia and selected European coun-
tries regarding launch in those markets.
Due to competitive reasons, the manu-
facturer has requested that it is not
named until the products are launched.
The reason is that the manufacturer con-
siders Zonase X(™) a novel and highly in-
novative technology.
Zonase X™ is a natural and bio active in-
gredient to the cosmetics and skin care
industry that adds moisture and increas-
es the renewal of skin in an effective and
gentle way. Zonase X™ derives from the
hatching fluid of salmon and contains
tiny patented components such as the
enzyme Zonase™.
Zonase™ is a non toxic, stable and spe-
cific enzyme which is why Zonase™ is
unique. The enzyme has very attractive
qualities; it gently exfoliates the dead
cells leaving the living cells untouched.
»ABT is very satisfied that we have fi-
nalized this Supply Agreement with a
highly regarded manufacturer of skin
care products. Since we first signed an
NDA in November 2009, we have in close
collaboration developed a number of ex-
citing products demonstrating the wide
application of Zonase X(™)« says Thor A.
Talseth, Executive Chairman of ABT.
■
SOFW-Journal | 136 | 11-2010
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