Amino Acid Complex

Amino Acid Complex,

”nutrients for skin”,

is a complex of amino acids, blended in the same or similar composition as

they are composed in the stratum corneum, that maintain the skin

moisturized, provide nutrients to the skin cells, proliferate skin fibroblasts

and prevent the skin from aging.

Amino Acids & ProteinsProteins are linear polymers whose monomeric building blocks are

“Amino Acids”

Central carbon (tetrahedral and chiral)

“R” functional group (the amino acid “side chain”)

Amino group and carboxylic acid group

Chemically linked Amino acid monomers to form proteins.

Types& structures of Amino Acids

Common amino acids20 common amino acids composed of C, H, O, N and S atoms.

Uncommon amino acidsHydroxylysine and hydroxyproline that are found in the protein collagen.

A common side chain reaction involving cysteineCystine

Types& structures of Amino Acids

NMF : all

Components of Collagen & Elastin : glycine, proline, alanine

Composing skin cell : aspartic acid, tyrosine, serine

Cell proliferation : aspartic acid, tyrosine, serine

Matabolism activation : aspartic acid, serine, proline

Removing freckles : cysteine

UV block : histidine, tryptophan, phenylalanine

Glutathion synthesis : glutamic acid, cysteine, glycine

Immune system enhancement : glutamine

Hair : arginine, histidine

Amino acids & Skin

The skin consists of several layers and the most important layer among the layers to

protect the body and maintain elastic and moisturized skin from the outside environment is

the epidermis.

The epidermis consists of stratum corneum, stratum granulosum, stratum spinusum and

stratum basale and maintaining proper water content on the stratum corneum is very

important to keep the skin elastic and supple, and NMF such as amino acids and

polysaccharides in the skin play a very important role in maintaining water contents on the

stratum corneum.

Damaged skin barrier

Weak Keratinocyte

Losing of moisture

Strong Keratinocyte

Recovered skin barrier with moisture

Amino Acids & Skin Moisturizing

1.2

1.5

3.2

3.4

6.9

Content(%)

halfcystein

Histidine

Phenylalanine

Tyrosine

Leucine

Amino acid

2.7Isoleucine12.6Glutamin acid

1.1Methionine13.6Serine

3.0Valine3.0Threonine

4.4Alanine7.9Aspartic acid

24.5Glycine3.8Arginine

3.0Proline4.2Lysine

Content(%)Amino acidContent(%)Amino acid

Table 1. Amino Acid composition in Stratum Corneum

The amino acid composition in the stratum corneum is as shown in Table 1 and the highest content

among the amino acids is Glycine. (Baden et. al, Biochem. Biophysics. Acta, 322, p269-278, 1973).

Amino Acids & Skin Moisturizing

Maintaining skin elasticity.

As we approach our 30s, our body's production of collagen and elastin (two substances that

give the skin firmness and elasticity) begins to slow down. The collagen and elastin also begin

to loosen and unravel which results in skin that sags and has poor elasticity.

Collagen synthesis is one of the most important activities of amino acids. Proteins account for

16~20% of our body weight and collagen accounts for a third of the proteins. Among amino

acids Glycine, Proline, Alanine are the raw materials of collagen.

Elastin

Collagen

Amino Acids & Skin Elasticity

Cell proliferation

Proteins are fundamental substances for growth, development, maintenance and repair

of all cellular tissues in the body and are used to produce hormones, enzymes, antibodies

and neutrotransmitters, and amino acids are the most basic components composing

these proteins.

Amino acids, as a source of energy supply to the skin and body, supply nutrients to each

cell thus and so proliferate fibroblasts, and promote collagen and elastin synthesis in the

skin that gives anti-aging effect.

Amino Acids & Cell proliferation

De-pigmentation

Cells called melanocytes are found in the transitional layer between the epidermis and dermis. These skin cells produce a brown-black skin pigment called melanin, which helps to protect against the damaging rays of the sun and to determine skin coloring. Melanin is discharged with

the metabolism of the skin and the skin color returns as time passes. However, as a person ages, the metabolism becomes slower and melanocytes often proliferate, forming concentrated clusters that appear on the surface as small, dark, flat or dome-shaped spots, which are usually harmless moles or liver spots.

Among amino acids, Aspartic acid, Tyrosine and Serine that

become skin cell as well as components of the enzyme for cell

division, promote the metabolism.

Moreover, Cysteine inhibits the activity of Tyrosinase and Tyrosine, Histidine, Phenylalanine and Tryptophan block UV that is one of the main reasons of melanin formation.

Amino Acids & Pigmentation

Moisturizing

Cell proliferation

TIMP-1 mRNA(Elasticity)

Anti- Aging

Effects of Amino acid complex

Amino acid complex

Method

Moisturizing effect of Amino acid complex

Amino Acid Complex

Desiccator Conditions

R.H. 50% , 35℃

Filled with CaCl2

Measuring weight change

0

20

40

60

80

100

120

0 3 24 29

Time(Hours)

Wat

er c

onte

nts(

%)

D.I water

1% hyaluronic acid

Amino acid complex

Result

Method

Continuous Moisturizing effect (In Vivo)

Result

Measuring moisture content

Corneometer CM825

(Courage & Khazaka, Germany)

R.H. : 45±5%

Temperature : 23±2℃

Application : 2mg/cm2 of cream containing 5% of Amino Acid Complex for 3 days

The changes in moisture content

15

20

25

30

35

0 1 2 3 6Day

Mo

istu

re v

alu

e

Treated site Untreated site

Method

Desquamation reduction effect (In Vivo)

Result

Measuring desquamation score

Visioscan VC98

(Courage & Khazaka, Germany)

R.H. : 45±5%

Temperature : 23±2℃

Application : 2mg/cm2 of cream containing 5% of Amino Acid Complex for 3 days.

Change of the desquamation score

0

0.5

1

1.5

2

2.5

3

3.5

4

0 1 2 3 6Day

Desquam

ation s

co

r

Treated site Untreated site

Pictures of In Vivo testTest

0 Day 3 Day 6 Day

0 Day 3 Day 6 Day

Control

Volunteer #1

Pictures of In Vivo testTest

0 Day 3 Day 6 Day

0 Day 3 Day 6 Day

Control

Volunteer #2

Pictures of In Vivo testTest

0 Day 3 Day 6 Day

0 Day 3 Day 6 Day

Control

Volunteer #3

Pictures of In Vivo testTest

0 Day 3 Day 6 Day

0 Day 3 Day 6 Day

Control

Volunteer #4

Pictures of In Vivo testTest

0 Day 3 Day 6 Day

0 Day 3 Day 6 Day

Control

Volunteer #5

Method

5x 104/well of HaCa-T cells with DMEM containing 10% FBS were cultured in

24well plate for 24 hours at 37℃, 5% CO2, and different concentrations of

Amino acid complex were added. The mixture was incubated for 72

hours at 37℃, 5%CO2 and 1ml of MTT(0.33mg/ml) was added and the

mixture was incubated for 4 hours at 37℃. MTT was removed and 1ml of

DMSO was added and the absorbance was measured at 570nm.

Cell proliferation of Amino acid complex

MTT is reduced to formazan by NADH in mitochondria

of viable cell, and has the maximum extinction at 570

nm wavelength.

Result

Cell Proliferation

50

60

70

80

90

100

110

120

5 9 23 33

Concentration(%)

Cel

l via

bilit

y(%

)

1 3 5 7

Cell proliferation of Amino acid complex

TIMP-1 mRNA Expression

Method

1×106 of fibroblasts were cultured in 100mm dish and incubated for 24 hours at 37℃. Different

concentrations of Amino acid complex were added and incubated for another 24 hours.

After removing the cell medium, 1ml of Trizol was added and RNA was isolated and quantified

according to the RNA Isolation Method by Invitrogen Co and RT-PCR was conducted with All-in-one

RT-PCR kit according to RT-PCR kit manual. Primer and reaction conditions are as follows.

TIMP-1 (Tissue Inhibitor of Matrix Matallo – Proteinase 1)

sense : 5'-ATCCTGTTGTTGCTGTGGCTGATA-3'

antisense : 5'-TGCTGGGTGGTAACTATTTATTTCA-3'

Actin ( Control )

sense : 5'-GAGACCTTCAACACCCCAGCC-3'

antisense : 5'-GGCCATCTCTTGCTCGAAGTC-3'

Anti-aging effect of Amino acid complex

Result

Amino Acid Complex promoted expression of TIMP-1 mRNA that is an inhibitor of MMP-1.

Amino Acid Complex 7% showed similar effect with Vit C and EGCG.

Amino Acid Complex has the anti-aging effect by promoting TIMP-1 mRNA expression that is involved in inhibition of wrinkle formation.

< TIMP-1 > < Actin >

lane 1 : Control (water)lane 2 : Amino Acid Complex 5%lane 3 : Amino Acid Complex 7%lane 4 : Ascorbic acid 150uMlane 5 : EGCG (Epigallocatechin gallate) 25uM

Anti-aging effect of Amino acid complex

Amino acid complex

Epidermis

Dermis

Follicular

Sebaceous gland

Skin penetration of Amino acid complex

Skin penetration

According to the references, most ingredients penetrate into the skin through sebaceous glands

and follicular. As amino acids have small molecular weight, they easily penetrate into the skin.

Cosmetics

All kinds of cosmetics for moisturizing & anti-aging

Toiletries1. Hair Care (Shampoo, Rinse, Hair essence etc…)2. Body Care (Soup, Bath&Shower Gel, Body cream etc..)

Body

Recommended Dosage : 5 ~ 10 %

Skin Hands

Hair

Application of Amino acid complex

Lysine, Histidine, Arginine, Aspartic Acid, Threonine, Serine, Glutamic Acid, Proline,

Glycine, Alanine, Valine, Methionine, Isoleucine, Leucine, Tyrosine, Henylalanine,cysteine

Colorless clear liquid

Typical

Purple

1.330 ~ 1.370

3.5 ~ 5.5

0.990 ~ 1.030

Not more than 20ppm

Not more than 2ppm

0.9 ~ 2.5%

Not more than 100 cfu/mL

Not more than 10 cfu/mL

INCI Name

Description-Color

-Odour

Identification

-Amino acids

Refractive Index

pH (Neat)

Specific Gravity (d20/20)

Purity

-Heavy Metals

-Arsenic

Dry Residue (1g,105℃,3hr)

Microbiological Purity

-Total bacterial count

-Yeast & molds

SpecificationsAnalytical Tests

Specification of Amino acid complex

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Amino Acid Complex