who brings anti-age science

and a sacred plant together?

we do.

harmonianceTM biofunctional

Vincience Zeta Fraction

TM TM

biofunctionals Technology
 a natural biofunctional produced from
the whole, fresh, living Sacred Lotus
Presentation outline:
− description

− background

− Zeta Fraction™ Technology

− evaluation

− applications

2
description
 Sacred Lotus

Sacred Lotus (Nelumbo nucifera)

is an iconic plant revered in Asia
as a divine symbol of beauty. An
aquatic perennial plant, its roots
go into the soil of ponds, while
the leaves float on top of the
water surface, or remain well
above it. Its flowers, found on
thick stems, rise several
centimeters above the leaves.

4
 origin and positioning

Origin
• Sustainable and traceable, our Sacred Lotus plants are grown in a dedicated farm
specifically for use in harmoniance™.
• Using Zeta Fraction™ Technology, the whole plant including the flower, stem, leaf and
root are separated into fractions to preserve original cellular architecture and to
capture synergistic properties.
• Ashland’s proprietary Zeta Fraction ™ Technology provides the powerful comprehensive
benefits of fresh and living Sacred Lotus in an exclusive biofunctional unobtainable by
conventional extraction techniques.

Positioning
• Today, Sacred Lotus conveys a positive story and image for people over the entire
world due to its ancestral role in traditional Asian medicine.
• harmoniance™ is a comprehensive & multi-functional ingredient that targets the most
important attributes of skin appearance associated with skin aging, including hydration
and barrier function, skin laxity and appearance of wrinkles, drainage and body
contouring, skin tone.
5
 harmoniance™ : a complete approach to
anti-aging face and body care applications
Aging skin harmoniance™ total anti-age benefits
In vitro studies:
skin hydration/barrier function - Antioxidant activity (ORAC, DPPH)
- Inhibition of elastase activity (-25%)
- skin dryness ↑ - Hyaluronic acid increase (+48%)
- Increased filaggrin expression (+65%)
- Increased AQP3 expression (+22%)
- Increased barrier function (+85%)
skin laxity and appearance of wrinkles - Increased collagen I expression (+38%)
- Increased glycerol release (+73%)
- ECM production ↓ - Anti-inflammatory activity on PGE2 (-50%)
- contractile properties ↓ - Melanin control (-25% - tyrosinase, TRP-2, TRP-1)
- Melanin control (-80%, ex-vivo human skin)

drainage & body contouring

- Orange peel skin appearance ↑
Clinical studies:
- Vascular reponsiveness ↓ - Reduction in appearance of wrinkles (-20%)
- Skin hydration (+14%), TEWL (-8%)
- Skin softness (+25%)
- Drainage & body contour (-12mm on thigh)
skin pigmentation & tone
- Age spots ↑
6
background
nature and science of Lotus
 the unique properties of Sacred Lotus

Recent molecular research has shown

that Lotus may be a “living fossil” that Traditional iconic image of purity and beauty

existed more than 138 Million years

ago. Enduring
Thermoregulating
seeds
Researchers also reported that the flowers

Lotus plant has the rare ability to Perennial

Self-cleaning leaves
regulate its temperature within a (ultrahydrophobic
growth
“Lotus effect”)
narrow range (just as humans do). Thrives even in polluted
waters
The well known “Lotus effect” refers to
self-cleaning properties that are a
result of ultra-hydrophobicity exhibited
by the leaves surface.

8
 longevity

Under favorable circumstances Lotus seeds may

remain viable for hundreds of years, with the oldest
recorded Lotus germination being from that of seeds
1,300 years old recovered from a dry lakebed in
northeastern China.
An individual Lotus can live for over a thousand
years and has the rare ability to revive into activity
after stasis.
The genome of the Sacred Lotus was sequenced in
May 2013. Scientists sequenced more than 86
percent of the nearly 27,000 genes of the plant
“which is believed to have a powerful genetic
system that repairs genetic defects, and may hold
secrets about aging successfully”1.

1 “Genome of the long-living Sacred Lotus (Nelumbo nucifera Gaertn.)”, Ming

et. al. Genome Biology 2013, 14:R41

9
 purity and beauty

Revered as a divine symbol for more than 5,000 years, the Sacred Lotus is
a truly iconic plant.
Its emblematic flower growing into pure beauty out of the mud of its
origin has for centuries symbolized a spiritual promise to humanity. Lotus
has always had a deep religious meaning to Hindus and Buddhists in
particular, to whom the Lotus flower symbolizes beauty, purity and
divinity. In Hinduism the Sacred Lotus represents the sun, and is
associated with mother goddesses as a symbol of fertility.
It is also the national flower of India, Bangladesh and Vietnam.
Lotus, because of its universal significance, vehiculates an extremely
attractive story and image that matches perfectly the ideal attributes of
a comprehensive cosmetic product: purity, lightness, beauty, and
perfection.

10
 medicinal and cosmetic applications

Used as a food for over 7,000 years in Asia, Lotus is also associated with
health and healing: it is an important and potent plant in traditional
medicines in Asia and virtually all parts of the Lotus plant including
stamens, pollens, flowers, stems, roots, and leaves are used to alleviate
numerous conditions such as depression, heart problems, hypertension,
insomnia, etc.
Deriving from its ancestral traditional uses, Lotus extracts from different
parts of the plant have been identified as having potential benefits
relating to skin and cellular biology: antioxidant, skin hydration and skin
barrier function, skin tone and fairness, help reduce irritation, and
inflammation, help improve skin texture, and attenuate the visible effects
of aging, help improve body contouring (1).

(1) Nelumbo Nucifera (Lotus): A Review on Ethnobotany, Phytochemistry and Pharmacology

11
zeta fractionTM technology:
capturing the natural power of
fresh and living Sacred Lotus.
 zeta fraction™ technology: a novel approach
to obtain the benefits of natural ingredients

• Zeta Fraction™ technology is a proprietary, solvent-free and

sustainable process that captures the essence of fresh and living
plants and forms the basis for robust ingredients and claims.
• The technology selectively separates intracellular components from
fresh living plants.
• Targeted fractions are separated by specially designed equipment.
Broad frequencies of electromagnetic waves are finely tuned to
specifically impact colloidal properties of plants’ intracellular
components to initiate their targeted separation.
• Includes only naturally occurring substances. No external chemicals
are used in manufacturing process.

13
 a sustainable and traceable technology

• No solvents used in the process

• No residual solvent
• No negative environmental impact or waste from the
harvesting and fractionation processes
• All parts of the Lotus plant are utilized or returned to
nature
• All materials come from identified and traceable lands
• GMO-free control
• REACH exemption

14
 Distribution of Plant Cell Components
distribution
Among Core Zetaof plant cell components
Fractions
among core zeta fractions
cell wall
insensitive
to
cell wall
dehydration fraction plasma
membrane

chloroplast
membrane
fraction
mitochondria
sensitive
nucleus
to cytoplasm
dehydration
fraction
cytoplasm

serum cytosol
fraction
vacuole

15
 processing on a live basis

harmoniance™
whole fresh living Lotus plant intracellular content Treatment
serum fraction

bioactive complexes stable colloidal dispersion targeted destabilization

cell wall fraction membrane fraction cytoplasm fraction

16
 harmoniance™ analytical profile

Zeta Fraction™ technology, in processing the entire fresh and living plant of Nelumbo nucifera,
enables the unique composition of harmonianceTM: a genuine, comprehensive, and
multifunctional ingredient that for the first time captures the synergistic properties of the Sacred
Lotus.
% w/w in “as is” material
Quercetin 0.0687
Luteolin 0.0024
Isorhamnetin 0.0005
Quercitrin 0.0042

Chlorogenic acid 0.0003

Other phenolics 1.25

Sucrose 1.16
Fructose 0.53
Raffinose 0.04

Free amino acids 0.151

Potassium 0.4
Sodium 0.2
Magnesium 0.03
Silicon 0.0007

Other 1.46

17
 harmoniance™ constituents

- harmoniance™ serum fraction contains hydrophilic and amphiphilic

constituents of intracellular colloidal dispersion that are separated from the
living Sacred Lotus plant.
- Product composition represents intracellular content of the living plant.

- Major constituents are:

- Phenolic compounds (total) ~1.33-1.58% w/w including:
- Quercetin, Luteolin, Isorhamnetin, Quercitrin
- Their structural isomers (e.g. Kaempferol) – predominately in glycoside forms
Chlorogenic acid, etc.
- Water soluble carbohydrates ~2.29-2.54% w/w including:
- Raffinose, Fructose, Sucrose, and their structural isomers
- Electrolytes, microelements, free amino acids

18
evaluation
• In vitro results, a summary
• Skin hydration and barrier function
• In vitro & ex vivo studies
• Clinical study, skin moisturization
• Skin laxity and appearance of wrinkles
• In vitro & ex vivo studies
• Clinical study, appearance of wrinkles
• Drainage and body contouring
• In vitro studies, lipolysis
• Clinical study, slimming appearance
• Skin tone
evaluation:
in vitro results, a summary
 harmoniance™ explorative activities in vitro:
a summary

Assay Activity (“as is”)

1g = 31mg (R)-Trolox methyl
ORAC (antioxidant)
ether
Mitigate oxidation
1g quenches 26mg DPPH
DPPH (quenching)
radicals
Mitigate Sun-induced Prostaglandin E2 IC50 ≤ 0.05%
inflammation Sun-induced Interleukin 8 IC50 ≤ 0.005%
(keratinocyte cell
culture) SDS-Induced Interleukin 8 IC50 = 0.02%
Extracellular matrix Elastase IC50 = 0.33%
integrity
(enzyme inhibition) Matrix Metallopeptidase 3 IC50 ≤ 0.03%

21
evaluation:
skin hydration and barrier function
in vitro & ex vivo studies
 hyaluronic acid expression

Cultures: Ex vivo human skin (age of donor 26, breast) Product application: harmoniance™ at 0.5% & 1% for 48 h (2x/d)
(Placebo=PBS)
Evaluation: Staining of hyaluronic acid Quantification: Image quantification by Volocity* software

Placebo 0.5% harmonianceTM 1% harmonianceTM

Ep: ns Ep: +48%***

Der: +18%* Der: +22%**
***: highly significant; **: very significant; *: significant; ns: non significant with Student’s t-test compared to placebo condition; n=3 x20

harmonianceTM at 1% was associated with an observed significant increase in

hyaluronic acid expression in ex vivo studies.

23
 filaggrin expression
Cultures: Ex vivo human skin (age of donor 30, arm) Product application: harmoniance™ at 0,5% &1% for 48 h (2x/d)
(Placebo=PBS)
Evaluation: Immunostaining of filaggrin Quantification: Image quantification by Volocity* software

Placebo 0.5% HarmonianceTM 1% HarmonianceTM

+39%*** +59%***

+65%***
***: highly significant, **: very significant; *: significant with Student’s t test compared to placebo condition n=2 x20

harmonianceTM at 0,5% and 1% was associated with an observed

significant increase in filaggrin expression in ex vivo studies.

24
 AQP3 expression
Cultures: Ex vivo human skin (age of donor 30, arm) Product application: harmoniance™ at 0,5% and 1% for 48 h (2x/d)
(Placebo=PBS)

Evaluation: Immunostaining of AQP3 Quantification: Image quantification by Volocity* software

Placebo 0.5% HarmonianceTM 1% HarmonianceTM

+22%*** +33%***
***: highly significant, **: very significant; *: significant with Student’s t test compared to placebo condition n=3 x20

harmonianceTM at 0,5% and 1% was associated with an observed significant

increase in AQP3 in ex vivo studies.
25
 skin barrier function
Fluorescent dye penetration was used to evaluate SDS stress on skin barrier function.
Cultures: 3D reconstituted human epidermis Product application: harmoniance™ at 0,5% & 1% for 48 h (2x/d) at D17 after stress
(Placebo=PBS)

Stress: 0.15% SDS at D17 during 3 h Evaluation: Staining with lucifer yellow fluorescent dye

-73%*** -85%***

SDS stress + 0.5% SDS stress + 1%

SDS stress + Placebo harmonianceTM harmonianceTM
Placebo
***: highly significant, **: very significant; ns: non significant with Student’s t test compared to placebo condition n=3 x20

• SDS stress showed severe disruption of the barrier function

• harmonianceTM at 1% was associated with an observed 85% reduction in
barrier disruption suggesting it is highly effective in maintaining skin barrier
function against environmental stress and pollution.

26
evaluation:
skin hydration and barrier function
clinical study skin moisturization
 protocol

Objective: Clinical study to investigate the moisturizing properties of

harmoniance™
Study: Double blind study with 0.5% harmoniance™ against placebo
Volunteers: 20 (23 to 64 years old) with normal or dry skin
Application: 1 application of a cream containing harmoniance™ at 0.5%
at a dose of 2 mg/cm² on the forearms
Duration: 6 hours
Control visits: T0, T3h and T6h
Measurements: Skin hydration, TransEpidermal Water Loss (TEWL) and
Frictiometer
Statistical analysis: Student’s t-test (one-tailed) or Wilcoxon signed rank
test (one-tailed) are used depending on whether the data followed a
normal distribution or not.

28
 hydration result

A highly significant
improvement of skin
moisturization was
observed 3h and 6h after
application.

Treated sides Time Mean (AU) sem p % of change % of improved volunteers

Placebo 5.77 1.13

T3h-T0 0.0006*** 13.92% 85% (17/20)
0.5% harmoniance™ 9.70 0.82
Placebo 5 0.92
0.5% T6h-T0 0.0004*** 14.50% 90% (18/20)
9.1 0.92
harmonianceTM
***: Highly significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal
distribution or not; mean +/- sem n=20
% of change = 100 * [(DxBiofunctional - D0Biofunctional) - (DxPlacebo - D0Placebo)] /((D0Biofunctional + D0Placebo) / 2)
29
 TEWL result

A highly significant
reduction of the TEWL was
observed at 3h and 6h
after application.

Mean % of improved
Treated sides Time sem p % of change
(g/m2h) volunteers
Placebo -0.93 0.22
T3h-T0 0.0001*** -7.55% 90% (18/20)
0.5% harmoniance™ -1.80 0.26
Placebo -0.71 0.17
T6h-T0 <0.0001*** -7.76% 87.5% (14/16)
0.5% harmonianceTM -1.60 0.20

***: Highly significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal
distribution or not; mean +/- sem n=20
% of change = 100 * [(DxBiofunctional - D0Biofunctional) - (DxPlacebo - D0Placebo)] /((D0Biofunctional + D0Placebo) / 2)

30
 frictiometer result

A highly significant
improvement of skin
softness was observed
3h and 6h after
application.

Treated sides Time Mean (AU) sem p % of change % of improved volunteers

Placebo 19.82 2.11

T3h-T0 0.0002*** 25.73% 90% (18/20)
0.5% harmoniance™ 26.12 2.53
Placebo 16.58 2.71
T6h-T0 0.0017*** 25.64% 85% (17/20)
0.5% harmonianceTM 22.86 2.86

***: Highly significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal
distribution or not; mean +/- sem n=20
31 % of change = 100 * [(DxBiofunctional - D0Biofunctional) - (DxPlacebo - D0Placebo)] /((D0Biofunctional + D0Placebo) / 2)
 conclusion

Three hours and 6 hours following the application of cream formulated

with 0.5% harmoniance™, a significant improvement in skin moisturization
is observed, compared to placebo.
This improvement is characterized by a significant reduction of TEWL,
indicating that harmoniance™ helps to restore skin barrier function.
Moreover, the frictiometer results at both times indicate that
harmoniance™ promotes the improvement of skin softness, compared to
the placebo sides.

32
Evaluation:
skin laxity and appearance of
wrinkles
in vitro & ex vivo studies
 collagen I expression

Cultures: Adult human fibroblasts (age of donor 53) Product application: harmoniance™ at 0.01% for 48 h (2x/d)
Evaluation: Immunostaining of collagen I Quantification: Image quantification by Volocity* software

Untreated 0.01% harmonianceTM

+38%***
***: highly significant with Student’s t test compared to placebo condition; n=3 x20

harmonianceTM at 0.01% was associated with an observed increase

in collagen 1 expression, in vitro.

34
 elastic fibers after uv stress
Cultures: Ex vivo human skin (age of donor 30, breast) Product application: harmoniance™ at 1% for 48 h (2x/d) (Placebo=PBS)
Stress: 5 J/cm² UVA + 150 mJ/cm² UVB Evaluation: Van Gieson staining

Elastic fibers: in black color

Collagen: in pink color

UVA+UVB harmonianceTM + UVA + UVB

A better organization of elastic fibers was observed when the skin was
pretreated with harmonianceTM at 1%, in ex vivo studies.
35
evaluation:
skin laxity and appearance of
wrinkles
clinical study
 protocol
Objective: Clinical evaluation of harmoniance™ activity on wrinkles appearance
Study: Double blind study with 0.5% harmonianceTM against placebo
Volunteers: 18 women (42 to 68 years old)
Application: 2 applications per day (morning & evening) of a cream containing
harmonianceTM at 0.5% concentration
Dose: 2 mg/cm² on the face
Duration: 8 weeks
Control visits: D0, D28 and D56
Measurements:
• Color pictures
• Pictures of the silicone replicas of the crow feet area
• Volunteer evaluation
• DermaTOP* pictures and analysis of the crow feet area
Statistical analysis: Student’s t-test (one-tailed) or Wilcoxon signed rank test (one-tailed)
were used depending on whether the data followed a normal distribution or not
37
 results: number of wrinkles

% of improved
Treated sides Time Mean sem p % of change
volunteers
Placebo -0.22 0.62
D28-D0 0.5ns - -
0.5% harmoniance™ -0.22 0.59
Placebo -0.06 0.55
D56-D0 0.0088** -20.26% 66.67% (12/18)
0.5% harmonianceTM -1.78 0.67
ns: not significant, **: very significant with Student’s t-test or Wilcoxon test depending on whether the data followed a
normal distribution or not; mean +/- sem n=18
% of change = 100 * [(DxBiofunctional -D0Biofunctional) - (DxPlacebo-D0Placebo)] /((D0biofunctional +D0Placebo)/2)

38
 results: volume of wrinkles

% of improved
Treated sides Time Mean (mm3) sem p % of change
volunteers
Placebo 0.03 0.11
D28-D0 0.0247* -13.72% 61.11% (11/18)
0.5% harmoniance™ -0.15 0.1
Placebo -0.01 0.1
D56-D0 0.0406* -11.68% 66.67% (12/18)
0.5% harmonianceTM -0.17 0.07

*: significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal distribution or
not; mean +/- sem n=18
% of change = 100 * [(DxBiofunctional -D0Biofunctional) - (DxPlacebo-D0Placebo)] /((D0biofunctional +D0Placebo)/2)
39
 results: area of wrinkles

Mean % of improved
Treated sides Time sem p % of change
(mm²) volunteers
Placebo 1.64 2.19
D28-D0 0.0256* -16.77% 61.11% (11/18)
0.5% harmoniance™ -1.99 1.34
Placebo 1.55 2.2
D56-D0 0.0152* -21.67% 72.22% (13/18)
0.5% harmonianceTM -3.14 1.5
*: significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal distribution
or not; mean +/- sem n=18
% of change = 100 * [(DxBiofunctional -D0Biofunctional) - (DxPlacebo-D0Placebo)] /((D0biofunctional +D0Placebo)/2)
40
 results: rugosity
0.5% harmoniance™ treated side

D0 D56

Treated sides Time Mean sem p % of change % of improved volunteers

Placebo 0.0036 0.0054

D28-D0 0.0124* -6.90% 72.22% (13/18)
0.5% harmonianceTM -0.009 0.0053
Placebo -0.0042 0.005
D56-D0 0.0243* -4.40% 66.67% (12/18)
0.5% harmonianceTM -0.0122 0.0035
*: significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal distribution or not;
mean +/- sem n=18
% of change = 100 * [(DxBiofunctional -D0Biofunctional) - (DxPlacebo-D0Placebo)] /((D0biofunctional +D0Placebo)/2)

41
 color pictures of crows feet:
(volunteer 16; age of donor 55)
Placebo 0.5% harmoniance™

Color pictures show

less visible wrinkles
and a smoother skin
D0

surface on the side

treated with
harmonianceTM 0;5%
compared to the
placebo-treated side
D56

42
 color pictures of crows feet:
(volunteer 1; age of donor 57)
Placebo 0.5% harmoniance™
D0
D56

43
 DermaTOP pictures:
(volunteer 18; age of donor 42)
Placebo 0.5% harmoniance™

harmonianceTM 0;5% is
associated with less
visible wrinkles and
D0

with a smoother skin

surface observed on 3D
dermatop images.
D56

44
 2D representation:
(volunteer 5; age of donor 52)
Placebo 0.5% harmoniance™

Results suggest that the

application of a cosmetic
D0

formulation containing
harmonianceTM at 0.5%
may help decrease
parameters of roughness
(depth of wrinkles
D56

correlated with intensity of

red color).

45
 3D representation:
(volunteer 5; age of donor 52)
Placebo 0.5% harmoniance™

harmonianceTM at 0.5% is
associated with an
D0

improvement in skin relief

after 56 days of
application.
D56

46
 results of Volunteer self evaluation

After one and two months of application, respectively, the volunteers

answered the following question:
− On which side of your face did you see an improvement?
− (Improvement means: wrinkle faded, decrease of signs of age, skin
appears more firm or more tonic, a better complexion and a more
radiance skin)
At D28, 50% observed an improvement of their skin on the
harmonianceTM side; 28% on the placebo side; and 22% had no opinion.
At D56, 67% of volunteers observed an improvement of their skin on the
harmoniance™ - treated side; 28% on the placebo treated side; and 5%
had no opinion.

47
 conclusion

A 2-months clinical study against placebo evidenced that

harmoniance™ is associated with significant improvements in:
− The appearance of wrinkles (number, volume, area)
− Skin roughness (Rz)
Improvements were significant and visible on color pictures, 2D and 3D
representations, confirming the positive effect of harmonianceTM
application on the visible signs of aging.
Under these conditions, harmonianceTM formulated at 0.5% helps
decrease significantly the visible signs of aging including the
appearance of wrinkles.

48
evaluation:
drainage and body contouring
in vitro studies, lipolysis
 in vitro evaluation of lipolysis
Cultures: 3T3-L1 Product application: harmoniance™ at 0.01% for 48 h (2x/d) at D12 of differentiation
Positive control: Caffeine at 2 mM for 48 h (2x/d) Evaluation: Glycerol release

Evaluation of harmonianceTM on lipolysis (hydrolysis of

triglycerides into glycerol and fatty acids)

+73%*

+31%*

Harmoniance 0,01%

*:significant with Student’s t test compared to placebo condition; mean ± sem; n=2

harmonianceTM at 0.01% was associated with an observed enhancement of the

glycerol release.
50
evaluation:
drainage and body contouring
clinical study, slimming appearance
 protocol

Objective: Clinical study to evaluate the effect of harmoniance™ on drainage and body
contouring.
Study: Double blind study with 0.5% harmonianceTM against placebo
Volunteers: 20 women (23 to 48 years old)
Application: 2 applications per day (morning & evening) on the thighs. Dose: 2 mg/cm²
Duration: 8 weeks
Control visits: D0, D28 and D56
Measurements:
• centimeter measurement of circumference of the upper and lower part of the thigh (5cm
above knees).
• Clinical and volunteer evaluation

Statistical analysis: Student’s t-test (one-tailed) or Wilcoxon signed rank test (one-tailed) are
used depending on whether the data followed a normal distribution or not

52
 results: centimeter measurement of
thigh circumference
• Upper part of the thigh:
Mean % of improved
Treated sides Time sem p % of change
(cm) volunteers
Placebo -0.04 0.109
D56-D0 0.0056** -0.88% 75% (15/20)
0.5% harmoniance™ -0.55 0.14

**: very significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal distribution or not; mean +/- sem n=20
% of change = 100 * [(DxBiofunctional - D0Biofunctional) - (DxPlacebo - D0Placebo)] / ((D0Biofunctional + D0Placebo) / 2)

• Lower part of the thigh:

% of
% of
Treated sides Time Mean (cm) sem p improved
change
volunteers
Placebo -0.41 0.128
D56-D0 0.0012*** -1.83% 90% (18/20)
TM
0.5% harmoniance -1.18 0.187
***: highly significant with Student’s t-test or Wilcoxon test depending on whether the data followed a normal distribution or not; mean +/- sem n=20
% of change = 100 * [(DxBiofunctional - D0Biofunctional) - (DxPlacebo - D0Placebo)] / ((D0Biofunctional + D0Placebo) / 2)

A very significant decrease of thigh circumference was observed

after 2 months application of the cream containing harmonianceTM.

53
 results: volunteer assessment
At D28 and D56, volunteers were asked two questions:
− Which thigh is the thinnest?
− On which thigh is skin more pleasant to touch?(no palpable of padded
skin, smooth and soft skin)?

After two months of

application, sixty percent
of volunteers reported
that the thigh treated
with 0.5 % harmoniance™
- containing cream
thinner and 55 % found
that skin touch was more
enjoyable.

54
 conclusion

Application of a cream containing harmoniance™ at 0.5%

was observed to be associated with a significant decrease
of the thigh circumference, compared to placebo.
Volunteer self assessment highlighted the perception of a
slimmer and more pleasant touch on the harmoniance™ -
treated side.
The present clinical study supports the potential of
harmoniance™ to help promote slimming and draining
through its beneficial effect on thigh circumference.

55
 representative thigh pictures after 56
days of treatment: volunteer 8

0.5% harmoniance™ Placebo

56
 representative thigh pictures after 56
days of treatment: volunteer 17

0.5% harmoniance™ Placebo

Age of donor, 29
57
evaluation:
skin tone
 evaluation on skin tone
Cultures: Ex vivo human skin (age of donor 52, breast) Product application: harmoniance™ for 48 h (2x/d)
Evaluation: Fontana Masson staining Quantification: Image quantification by Volocity* software

Placebo Kojic acid -67%

***

0.5% NS 1% -78% 3% -91%

harmonianceTM harmonianceTM *** harmonianceTM ***

***: highly significant, **: very significant; ns: non significant with Student’s t test compared to placebo condition n=3 x20

harmonianceTM at 1% and 3% was associated with an observed significant

reduction in melanin content.
59
applications
 harmoniance™, cosmetic applications

− Multifunctional “total” age defying face care day cream

− Face care formulation to brighten skin tone
− Sunscreen formulations
− Formulation with sustainable and green message
− Body and face care formulations for skin moisturization
− Formulation to address the appearance of sensitive skin
− Body contouring gels to reduce the appearance of orange peel
− Possible association with other biofunctionals to provide additional protection from the
environment such as GP4G SP™ biofunctionals (solar and thermal shocks) or Elixiance™
biofunctional (air pollution)

Recommended use level: 0.5 to 1% (clinically tested at 0.5%)

INCI: Nelumbo Nucifera (Lotus) extract
Preservative system: Potassium Sorbate, Sodium Benzoate

harmonianceTM INCI name is published on the Inventory of Existing Cosmetic Ingredients in

China – IECIC (2014)
61
 harmoniance™ safety/tox profile

• it is not a skin irritant based on in vitro skin irritation test

(reconstructed human epidermis test) and 48-hour occlusive
human patch testing
• it is not an eye irritant based on Hen's Egg Test – chorioallantoic
membrane (HET-CAM) test and in vitro eye irritation test
(reconstituted human corneal epithelium test)
• it has no phototoxic and no genotoxic potential in in vitro
phototoxicity study (3T3 Neutral red uptake phototoxicity assay)
and in vitro genotoxicity study (bacterial reverse mutation
"Ames" assay), respectively
• it is not a skin sensitizer considering that no irritation or
sensitization was observed in human repeat insult patch (HRIPT)
testing conducted with more than 200 volunteer panelists

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 disclaimer
The information contained in this presentation and the various products described are intended for use only by persons
having technical skill and at their own discretion and risk after they have performed necessary technical investigations,
tests and evaluations of the products and their uses. This material is for informational purposes only and describes the
scientific support for the use of the products described herein as an ingredient in cosmetic products intended to
enhance appearance and other cosmetic benefits or to enhance performance of an end product. Certain end uses of
these products may be regulated pursuant to rules governing medical devices or other regulations governing drug
uses. It is the purchaser’s responsibility to determine the applicability of such regulations to its products. While the
information herein is believed to be reliable, we do not guarantee its accuracy and a purchaser must make its own
determination of a product’s suitability for purchaser’s use, for the protection of the environment, and for the health and
safety of its employees and the purchasers of its products.

Neither Ashland nor its affiliates shall be responsible for the use of this information, or of any product, method, formulation,
or apparatus described in this brochure. Nothing herein waives any of Ashland’s or its affiliates’ conditions of sale, and no
statement, information and data is to be taken as a guarantee, an express warranty, or an implied warranty of
merchantability or fitness for a particular purpose, or representation, express or implied, for which Ashland and its affiliates
assume legal responsibility. We also make no warranty against infringement of any patents by reason of purchaser’s use
of any information, product, method or apparatus described in this presentation.

The testing information (the “Testing Information”) has been gratuitously provided by Ashland. The Testing Information is
based on many factors beyond Ashland’s control, including but not limited to, the conditions prevailing when the testing
was conducted, and in some cases, is based on data generated with development samples of the Active
Ingredient. Although it is intended to be accurate, ASHLAND DISCLAIMS ANY AND ALL LIABILITY, EITHER EXPRESS OR
IMPLIED. The Testing Information is confidential or proprietary to Ashland, and may not, except as provided below, be
disclosed to any third party. You may not make commercial use of the Testing Information, or make claims with respect
to your products based on the Testing Information, without the written agreement of Ashland covering such use.
® Registered trademark, Ashland or its subsidiaries, registered in various countries
™ Trademark, Ashland or its subsidiaries, registered in various countries
* Trademark owned by a third party
© 2017, Ashland

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