DI-NACERC-02

Nano Ceravit C

Ctra. Alpedrete, 6 – 28400 Collado Villalba, Madrid, España Tel. +34- 918 510 514. Fax +34 918 510 051. www.cobiosa.com

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INCI name

Glycerin (and) Ascorbyl Tetraisopalmitate (and) Ceramides (and) Pentylene Glycol

Description

NANO CERAVIT C consists primarily in Glycoceramides and Ascorbyl Tetraisopalmitate and is characterized by its exceptional galenic form: nanoscopic liposomes.

This has been achieved by using a special unique technique and it is able to have an increased efficacy on the skin. Liposome is an artificial microscopic vesicle whose membrane is made of one or more lipid layers. Thanks to the bio-affinity of the liposome membrane and the membrane of the skin cells, the vitamin C is effectively delivered where it is needed. The nanoscopic particle size of NANO CERAVIT C provides a high penetration of the active in the skin.

With this active ingredient, it is possible to have a highly concentrated dose of oil soluble vitamin C, microencapsulated in a ceramide nano-liposome.

DI-NACERC-02

Nano Ceravit C

Ctra. Alpedrete, 6 – 28400 Collado Villalba, Madrid, España Tel. +34- 918 510 514. Fax +34 918 510 051. www.cobiosa.com

2/7

The outstanding penetration of Vitamin C and the bio availability of the Ceramides in the skin has never being achieved before.

Effects in Cosmetic Products

At NANO CERAVIT C we have associated 2 effective active ingredients in a nanoscopic size liposome:

- Oil soluble Vitamin C ( ±20,00 %)

- Glycoceramides (±5,00%) Each one plays an important role in skin treatments: Vitamin C: Vitamin C has many claims as cometic ingredient. Nevetheless it is very unstable at cosmetic formulations because its oxidation and decoloration problems. Ascorbyl Tetraisopalmitate is an oil-soluble Vitamin C derivative. It is stable at high temperatures and it has a very good solubility in oils. It has excellent percutaneous absortion and it is broken into free VITAMIN C in the skin, to perform physiological functions. Vitamin C is widely used in cosmetic products thanks to its ability to build collagen. Collagen production decreases with age and at premature aged skins. It has been clinically proven that Vitamin C can stimulate collagen production by the skin. The skin uses an enzyme called proline to determine when more collagen needs to be produced. Vitamin C can mask the presence of the enzyme so the skin, unable to detect proline, starts producing more collagen. The final result is a firmer and more youthful skin. Besides this important effect, Vitamin C is a Free-Radical scavenger that has a major anti-oxidation effect. It can also reduce hyper-pigmentation and age spots in the skin as it can inhibit formation of excess melanin. Glycoceramides: Ceramides are of most importance to maintain a good barrier function at the stratum corneum. They reinforce, maintain and restore the lipidic layers and perform as cement for he skin corneocites. The nanoscopic liposome is made out of pure ceramides, not phospholipides With Ceramides we will:

• Reinforce the skin’s barrier function

• Protect the skin against external aggressions

• Modulate the enzymatic activity (equilibrium integrity/degradation of corneodesmosomes)

• Restore the hydroretention capacity of the skin, reducing the TEWL

DI-NACERC-02

Nano Ceravit C

Ctra. Alpedrete, 6 – 28400 Collado Villalba, Madrid, España Tel. +34- 918 510 514. Fax +34 918 510 051. www.cobiosa.com

3/7

Dosage – Solubility – Processing

A- DOSAGE: From 0,50 to 3,00 % B- SOLUBILITY: Water dsipersable, which is a remarkable property when working with ceramides and oil soluble vitamin C. C- PROCESSING: It would be preferably incorporated into cosmetic preparations during the finishing process, at the cooling phase at a temperature of not over 40

0C.

Analytical Data

- APPEARANCE: sirop, light yellow colored. - Peroxyde index: <1 - Refractive index (at 20ºC): 1,466 – 1,472 - PRESERVATIVES: none - MICROBIOLOGY: Maximum 10 CFU/gram (not pathogenic) - TOLERANCE: Excellent - STORAGE: Preferably store under refrigeration. If not, avoid high temperatures (over 30

0C), dry and away from light.

If original container is opened, to avoid secondary microbiological contamination handle with special care and keep refrigerated.

DI-NACERC-02

Nano Ceravit C

Ctra. Alpedrete, 6 – 28400 Collado Villalba, Madrid, España Tel. +34- 918 510 514. Fax +34 918 510 051. www.cobiosa.com

4/7

Efficacy tests:

Several tests have been carried out on the effecs of Ascorbyl Tetraisopalmitate in the skin:

1- Promotion of Collagen Synthesis:

Proline involved in collagen synthesis was added to human dermal fibroblast with several concentrations of Ascorbyl Tetraisopalmitate (ATIP) and L-ascorbic acid as reference, and cultivated during 24 hours. After, collagen fractions were obtained and measured. ATIP significantly promoted collagen synthesis

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0 10 20 30

Concentration

Concentration (u mol/L)

Am

ou

nt

sy

nth

es

ize

d (

10

4 d

pm

/10

6

ce

lls

)

Control

L-ascorbic

ATIP

DI-NACERC-02

Nano Ceravit C

Ctra. Alpedrete, 6 – 28400 Collado Villalba, Madrid, España Tel. +34- 918 510 514. Fax +34 918 510 051. www.cobiosa.com

5/7

2- Singlet Oxygen Scavenging (anti free-radical activity): By scavenging singlet oxygen molecules, preventing it from oxidizing L-Dopa to dopachrome, we can avoid the oxidation of the skin cells, building an anti-free radical activity. In order to prove this, a mixture of Hematoporphyrin and L-dopa with Ascorbyl Tetraisopalmitate (ATIP) and L-ascorbic acid, was irradiated with UVB, in order to produce singlet oxygen and, subsequently, dopachrome. The amount of produced dopachrome was then determined through its absorbance. With this measured value, the scavenging activity of both ATIP and L-ascorbic acid could be calculated:

77

77,5

78

78,5

79

79,5

80

80,5

Sin

gle

t O

xyg

en

Scaven

gin

g

L-Ascorbic Acid

Ascorbyl

Tetraisopalmitate

UVB irradiation: 4 hours, Concentration: 2,5 X 10

-4 mol/L. Amount of dopachrome determined

by its absorbance at 475 mm

3- Inhibition of Melanogenesis: To show the ability of Ascorbyl Tetraisopalmitate to inhibit the melanogenesis, and thus to reduce the amount of melanin in the skin (lightening effect), several concentrations of ATIP were added to cultures human melanoma cells. After a cultive of 4 days, the amount of melanin produced was measured by observation of the color tone of each cell pellet. As it can be seen at the figure, ATIP inhibited the melanogenesis and the results were dose-dependent

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40

50

60

70

80

90

100

0 0,005 0,01 0,1 0,2

Melaninamount(%)

DI-NACERC-02

Nano Ceravit C

Ctra. Alpedrete, 6 – 28400 Collado Villalba, Madrid, España Tel. +34- 918 510 514. Fax +34 918 510 051. www.cobiosa.com

6/7

Particle size:

The particle size distribution of Nano Ceravit C was measured by using a laser diffraction type

particle size measurement apparatus (SALD-200J, Shimadzu Corp.).

Measuring condition:

Refractive index: 1.70 – 0.20 i

Dispersion media: water Result Integration value

Frequence value

Rela

tive q

ua

ntity

of p

artic

le s

eiz

e

DI-NACERC-02

Nano Ceravit C

Ctra. Alpedrete, 6 – 28400 Collado Villalba, Madrid, España Tel. +34- 918 510 514. Fax +34 918 510 051. www.cobiosa.com

7/7

The image of particles size of Nano Ceravit C was analyzed by transmission electron

microscopy (TEM).

TEM image (JEOL JEM-2000 EX (TEM) 100 kV)

Fig. 1&2 Negative staining by uranium acetate

Fig. 3 Negative staining by phosphotungstic acid