Transcript Anti oxidant - home.kku.ac.th

Anti oxidants for Cosmetics
by
Dr. Panvipa Krisdaphong
Dean
School of Cosmetic Science
Mae Fah Luang University
Anti oxidants
* Outer beauty: Topical Cosmetics
Shining healthy hair and a clear radiant complexion.
* Inner beauty: Oral Cosmetics
The beauty that shines from inside.
* Lasting beauty: Topical Cosmetics & Oral Cosmetics
Younger than your chronological age.
Aging
1. Genetic level – DNA
2. Cellular level - Free radical
External Source
Sun
X-ray
Pollutant
Smoke
Radical
Toxic-chemical
Diet
Drugs
Anti-oxidation
Radical Theory
1) Free radical source
Immune destroy
virus/bacterial
Activities enzyme
Energy
Anti-oxidation
2) Dangerous of Free radical
Damage to cells
Change protein structure
Damage to tissue
Protein synthesis error
Anti-oxidation
Free radical
Atom & molecule with one or more unpaired electron
- Reactive Oxygen Species (ROS)
1.
Superoxide radical (O•2)
2.
Hydrogen peroxide (H2O2)
3.
Hydroxy radical (•OH) **
1. Superoxide radical (O•2)
Attack in body : - Enzyme
- Cell membrane : “lipid peroxidation”
O•2 + Unsaturated fatty acid
lipid free radical
brack down to cell
2. Hydrogen peroxide (H2O2)
Attack in body : - Nucleus (attack DNA : protein cross link)
3. Hydroxy radical (•OH) **
** Most potent oxidant
** Major causes of the change we see in aging
Attack in body : - Enzyme
- Proteins (attack DNA : most injurious to body overall)
- Carbohydrates
- Lipid
All reactive oxygen species (ROS)
Mitochondria**
Cell membrane
Hydroxy radical
Peroxide
Superoxide
Lipid peroxidation
Peroxisome
Peroxide
Nucleus DNA attack
Cellular proteins
all Reactive Oxygen Species
Hydroxy radical
Peroxide
Cell
All reactive oxygen species (ROS)
Mitochondria**
Cell membrane
Hydroxy radical
Peroxide
Superoxide
Lipid peroxidation
Peroxisome
Peroxide
Nucleus DNA attack
Cellular proteins
all Reactive Oxygen Species
Hydroxy radical
Peroxide
Cell
Mitochondria
Function : - Produce energy by oxidation.
- Produce free radicals
- 1º sites for free radical damage
Ref : The free radical theory of aging , Nathan C. Nelson, Department of Physics, Ohio State University, Columbus,
OH 43201
Source of oxidation
Anti oxidants for Cosmetic
•
Topical Cosmetic
•
Oral Cosmetic
Topical Cosmetic
• Pomegranate
• Giant curcuma
• Rhubarb
Pomegranate
Botanical name
Family name
Thai name
Chinese name
Part use
Origin
Compound
:
:
:
:
:
:
:
Punica granatum L.
PUNICACEAE
Tab-tim (ทับทิม)
Shi Liu Pi
Fruits
Southwest Asia.
Polyphenolic compound
Traditional used :
1. Diarrheal remedy
2. Anthelmintic remedy
Action :
1. Anti-oxidant
2. Anti-bacterial
Antioxidant activity
Antioxidant activity of pomegranate fruit extract and its
anthocyanidins: delphinidin, cyanidin, and pelargonidin
Method
:
In vitro (rat brain)
Examined
:
ESR technique* with spin trapping
(Free radical scavenging activities)
Result
:
- Pomegranate extract can exhibited scavenging activity
against .OH and O2 . -.
- Anthocyanidins scavenged O2·- in a dose-dependent
- The ID50 values** of delphinidin, cyanidin, and
pelargonidin were 2.4, 22, and 456 ų M
*ESR = Electron Spin Resonance (found that superoxide anion free radicals in mitochondria)
** ID50 values = dose of inhibition
Ref : Noda Y, Kaneyuki T, Morl A and Packer L., Antioxidant activity of pomegranate fruit extract and
its anthocyanidins: delphinidin, cyanidin, and pelargonidin. J Agric Food Chem. 2002 Jan 2 ; 50(1) : 166-71.
Antioxidant activity
Pomegranate (Punica granatum L.) fruits are widely consumed
as juice (PJ). The potent antioxidant and anti-atherosclerotic activities of
PJ are attributed to its polyphenols including punicalagin, the major
fruit ellagitannin, and ellagic acid (EA)
Sample
Pomegranate Juice
Total Pomegranate Tannin
Punicalagin
Ellagic acid
Antioxidant activity
++++
+++
++
+
In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tanni
extract are enhanced in combination with other polyphenols as found in pomegranate juice. Seeram NP, Adams LS,
Henning SM, Niu Y, Zhang Y, Nair MG, Heber D. Center for Human Nutrition, David Geffen School of Medicine,
University of California, Los Angeles, CA 90095, USA. [email protected]
Antibacterial activity
Successive petroleum ether, chloroform, methanol and
waterextracts of Punica granatum Ž . were tested in vitro for their
antibacterial activity.
Method
Test on
:
:
In vitro (Agar dilution)
Staphylococcus aureus MTCC 737
Escherichia coli MTCC 723
Klebsiella pneumoniae MTCC 109
Proteus ulgaris MTCC 1771
Bacillus subtilis MTCC 441
Salmonella typhi MTCC 537
Ref : Antibacterial activity of Punica granatum D. Prashanth, M.K. Asha, A. Amit Microbiology
Laboratory, Research & Deelopment Centre, Natural Remedies Pt. Ltd., Plot No. 5B, Veerasandra
Indl. Area, Hosur Road, Bangalore 561 229, India
Result
: Antibacterial activity of extractives from Punica granatum
fruit *
Tested material
Minimum inhibitory concentration (mg/ml)
S. aureus E. coli
Petroleum ether extract
Chloroform extract
Methanol extract
Water extract
Chloramphenicol (Standard)
12
12
6
25
0.0025
K. pneumoniae P. ulgaris B. subtilis
12
12
12
50
0.005
12
25
12
Not active
0.005
12
12
1.5
12
0.005
12
6
6
25
0.005
S. typhi
12
12
12
25
0.0025
*All determinations were done in triplicate.
Conclusion :
All the tested extracts showed some antibacterial
activities which were significant for the methanolic extract, especially
against P. ulgaris** and B.subtilis***.
** P. ulgaris : (urinary tract infections, infant diarrhea, respiratory infections)
*** B.subtilis : A common soil bacterium (illness causing diarrhea, nausea, vomiting)
Cardiovascular
As cellular oxidative stress influences both scavenger receptormediated uptake of Ox-LDL and cellular cholesterol biosynthesis.
Method
: in vitro
- Effect of Pomegranate on macrophage oxidative status.
- Macrophages were incubated without (control) or with
increasing conc. of pomegranate for 90 min at 37 C.
Measurement
: Cellular oxidative stress following cell incubation.
- Cellular fluorescence was determined (cytometry analysis)
Result
: Pomegranate shown to reduces cholesterol accumulation.
- inhibit macrophage foam cell formation
- inhibit development of atherosclerotic lesions
Ref: Pomegranate juice flavonoids inhibit low-density lipoprotein oxidation and cardiovascular diseases: studies in
atherosclerotic mice and in humans. Aviram M, Dornfeld L, Kaplan M, Coleman R, Gaitini D, Nitecki S, Hofman A,
Rosenblat M, Volkova N, Presser D, Attias J, Hayek T, Fuhrman B.Lipid Research Laboratory, Technion Faculty of
Medicine, Rappaport Family Institute for Research in the Medical Sciences, Rambam Medical Center, Haifa, Israel,
31096. [email protected]
Cellular Oxidative Stress
(Mean Fluorescence Intensity)
Pomegranate reduces oxidative stress in macrophages
300
250
200
150
100
50
0
0
12
25
50
75
(Control)
Pomegranate Juice Concentration (mmol of total polyphenols/L)
Conclusion
: Demonstrates that upon in vitro incubation of
macrophages with Ppomegranate, cellular uptake of Ox-LDL and cellular
cholesterol biosynthesis were significantly reduced parallel to a significant
reduction in cellular oxidative stress.
These effects were possibly responsible for the observed
reduction in foam cell formation and atherosclerotic lesion attenuation by
Pomegranate consumption .
Application
 Skin care product :
- Anti aging products
- Anti bacterial products
Rhubarb
Botanical Name : Rheum Officinale Baill..
Common Name : Rhubarb
Part Used : Root, Rhizomes
Rhubarb Originated
This plant originated from the Asian such as
* Thailand
* China
ACTIVE INGREDIENTS
Anthraquinones
1,8-dihydroxyanthraquinone; R1=H, R2=H
Aaloe-emodin; R1=H, R2=CH2OH
Chrysophanol; R1=H, R2=CH3
Rhein; R1=H, R2=COOH
Emodin; R1=H, R2=CH3
Physicion; R1=OCH3, R2=CH3
The active ingredients in rhubarb are
anthraquinnone such as rhein
Analysis of HPLC
In Cosmetics
The Rhubarb extract is use as
whitening products
Antioxidant
Antioxidant Phenolic Constituents in Root of Rheum officinale
Method
: in vitro
Radical scarvenging activities (RSA)
Determine by : Sample + ABTS+ (radical compound)*
Detection by : Absorbance
* 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)
Ref:Cai Y, Sun M, Xing J, Corke H. Antioxidant phenolic constituents in roots of Rheum officinale and
Rubia cordifolia: structure-radical scavenging activity relationships.J Agric Food Chem 2004 Dec 29;52
(26):7884-90.
Antioxidant
Result
Hydroxyanthraquinone from roots of R. officinale
Name
Anthraquinone glycoside
Aloe emodin
Rhein
Emodin
R. officinale Crude extract
Content (%)
6.8
1.5
1.9
2.6
Alcoholic extract
Radical scarvenging activities
(mM)
0.172 ± 0.003
0.173 ± 0.001
0.174 ± 0.001
0.172 ± 0.002
88.6 ± 6.4
Ref:Cai Y, Sun M, Xing J, Corke H. Antioxidant phenolic constituents in roots of Rheum officinale and
Rubia cordifolia: structure-radical scavenging activity relationships.J Agric Food Chem 2004 Dec 29;52
(26):7884-90.
Tyrosinase inhibitory activity
Method
Measurement
Result
:
:
in vitro
Determination of tyrosinase inhibitory activity
was performed by the dopachrome method*
using L-DOPA as the substrate.
:
Rhubarb extract had 28.03 % tyrosinase inhibition**
at 0.5 mg/ml.
*Dopachrome = intermidiate substances in the melanin biosynthesis
**The potent tial tyrosinase inhibitor would show decrease dopachrome absorbtion
Evaluation of Rheum extract cream
for whitening products
1. Ethical committee to ask for Ethical Clarance
2. Seletion subjects
- Male or Female
- Age 20-35 years
- No history of skin disease such as eczema or psoriasis
- No use the whitening products belong 6 month
- No use the whitening products between the test
3. Study design
Dose
Often
Duration
:
:
:
3 % rhubarb extract cream
Twice daily
8 weeks
* Compare with cream base
4. Determination of skin condition
Before application :
After application :
Melanin value
Melanin value (Every 2 week
during the period of 8 weeks)
5. The in vivo results
Result
:Increase in parameter L*after application
Positive depigmenting effect
Conclusion :Rhubarb creams (3 % ww) significantly
decreased skin color intensity compare
with control
L*parameter
- For clearity;from dark to light
- If the L*parameter is high, the skin color is light.
Chromameter® measurement
“Evaluation of the color of the treated area”
Principle
“The chromameter converts colors into a digital code”
80
L* parameter
L : Clarity (dark to light)
78
76
55°
74
a : Green to Red spectrum
very light
72
70
68
b : Blue to Yellow spectrum
41°
light
66
64
62
medium
These parameters are
exploited through the
calculation of the Individual.
28°
60
58
dark
(dark)
56
54
52
ITA
very dark
50
0
2
4
6
8
10°
10 12 14 16 18 20 22 24 26 28
b* parameter
Typological Angle (ITA),
which defines the degree of
skin pigmentation
The higher L parameter and the ITA are, the lighter the skin
Calculation formulas
Variations () of the colorimetric parameters L*, b* and
the ITA° were calculated in arbitrary units according to the
following formula:
Variations () = Ti - T0
Ti : value of the colorimetric parameters on the treated zone
on D28 and D56
T0: value of the colorimetric parameters on the treated zone
on D0
Oral Cosmetic
•
•
•
•
Resveratrol
Black paper
Emblic
beta glucan
Resveratrol
Source :
Bread fruits
Function: • Anti-oxidant
• Anti-cancer
• Cardioprotective
• Potential cholesterol lowering effect
Dose:
30 - 50 mg day
Compound: Polyphenolic compound
Cardioprotective effect
• Inhibition of platelet aggregation
• Promotion of vasodilation by enhancing the
production of nitric oxide
• Inhibition of inflammatory enzymes
Ref 1: Ray PS, Maulik G, Cordis GA, et al. The red wine antioxidant resveratrol protects isolated rat hearts from
ischemia reperfusion injury. Free Rad Biol Med. 1999; 27:160-169
Ref2:Pace-Asciak CR, Hahn S, Diamandis EP, et al. The red wine phenolics trans-resveratrol and quercetin
block human platelet aggregation and eicosanoid synthesis: implications for protection against coronary heart
disease. Clin Chim Acta. 1995; 235:207-219.
Anti oxidation
Antioxidant (on blood platelet function)
It is a potent antioxidant, reactive oxygen species scavenger
and metal chelators.
Resveratrol reduces lipid peroxidation, oxidation and
nitration of platelet and plasma proteins.
Lipid
peroxidation
Nitration of
platelet
(-)
inhibit
Platelet recruitment
Thrumbus formation
Plasma
proteins
Oxidation of Resveratrol
Cardiovascular
disease
Inhibition of platelet aggregation
Method
:
Dose
Result
:
in vitro
induce platelet aggregation by collagen,
thrombin, and ADP*
10-1000 µmol/l resveratrol
Inducer
% Platelete aggregation
Control
Thrombin
ADP
Collagen
82.912.5
64.8  5.1
74.0  4.0
Resveratrol
Resveratrol
Resveratrol
Aspirin
(10 µmol/l)
(100 µmol/l)
(1000 µmol/l)
(10 µmol/l)
64.0 
55.2  6.2a
60.6  4.8a
43.6 
46.2  9.3b
47.2  7.1b
13.9a
6.3b
30.6 
15.5  2.3b
26.9  4.3b
5.2b
52.5  4.6b
48.8  6.3b
43.9  4.7b
ap<0.05, bp<0.01,
compared with control group.
*ADP : Adenosin diphosphate
Ref: Effects of red wine and wine polyphenol resveratrol onplatelet aggregationin vivo and in vitro ZHIRONG WANG1, YUANZHU HUANG1, JIANGANG
ZOU1, KEJIANG CAO1, YINAN XU1 and JOSEPH M. WU21Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University,
Nanjiang 210029, China;2Department of Biochemistry and Molecular Biology and Brander Cancer Research Institute,New York Medical College, Valhalla, NY
10595, USAReceived October 22, 2001; Accepted November 12, 2001
Vascular Protection
Resveratrol can protecting the vascular walls
from oxidation, inflammation, platelet aggregation,
and thrombus formation.
In this review, we focus on the mechanism of
resveratrol cardiovascular benefic effects.
Ref : Resveratrol: preventing properties against vascular alterations and ageing.Delmas D, Jannin B, Latruffe N.
University of Burgundy, Laboratory of Molecular and Cell Biology, Dijon, France.
Anti-cancer and Immune stimulating
Preliminary test are findings of some resveratrol-related
anti-cancer and immune-stimulating effects.
Method
:
in vitro studies
Conclusion
:
1. Resveratrol can inhibit tumor initiation,
promotion and progression.
2. inhibit DNA synthesis and ribonucleotide
reductase in mammalian cells.
Black paper
Botanical name
:
Piper nigrum Linn.
Family name
:
PIPERACEAE
Part used
:
Seed
Origin
:
Jungles of southwestern Asia
Antioxidant efficacy
Method
Subject
Duration
Result
:
:
:
:
Conclusion
:
In vivo
30 male rats.
10 weeks.
Supplementation with black pepper
or piperine lowered TBARS
And CD levels and maintained SOD, CAT,
GPx, and GSH levels to near those
of control rats.
Supplementation with black pepper or
the active principle of black pepper, piperine,
can reduce high-fat diet induced oxidative
stress to the cells.
Ref : Vijayakumar RS, Surya D, Nalini N.Department of Biotechnology, Annamalai Nagar, Tamilnadu, India.
Emblica officinalis
Botanical Name
Family name
Common name
Thai name
Part use
: Emblica officinalis
: EUPHORBIACEAE
: Emblic
: Ma-kham-pom
: Fruits
Description
It grows in tropical and subtropical parts of China,
India, Indonesia and Thailand
Traditional medicine : Treat broad spectrum of disorders
•Tonic
•Diuretic
•Rejuvenative
•Anti-tussive
•Astringent
•Expectorant
•Antipyretic
•Anti-diarrheal
Composition
•
Ascorbic acid
•
Flavonoids
•
Tannins
•
Alkaloids
•
Diterpene
•
Triterpene
•
Benzenoid (Gallic acid)
Scientifically published data
In vitro
In vivo
- Anti oxidant
- Ulcer protective activity
- Anti inflammatory
- Anti diabetic activity
- Anti bacterial
- Reduce lipid level
- Anti fungal
- Protective liver injury
- Anti viral
- Immuno-modulatory
- Anti mutagenic
- Anti-oxidant
Active Compound
OH
HO
HO
OH
HO
OH
HO
O
O
HO
O
OH
O
O
O
O
O
HO
C
HO
HO
R2
R1
HO
OH
Gallic acid
Hydrolyzable tannin
Anti-oxidation of Emblic
Environment
Emblic extract
Reactive Oxidation Species
(ROS) O•2, H2O2 , •OH
(-)
SOD, Catalase etc.
Damage of the cell
Free radicals
Oxidative Stress
Anti-oxidation
Scientifically published data
Activity
:
Anti-oxidant
Method
:
In vitro
Material
:
Methanolic extract
Result
:
The concentration needed for the
inhibition of hydroxyl radical
scavenging were 155.5 mg/ml,
and that for superoxide scavenging
activity were found to be 6.5 mg/ml
Ref : Sabu MC, Kunttan R. Antidiabetic activity of medical plant and its relation ship with their antioxidant activity.
J Ethanopharmacol 2002;81:155-60
Scientifically published data
Activity
Method
Test on
Dose
Result
:
:
:
:
:
Anti-oxidant
In vivo
rat liver
5-500 mg/ml
Emblic extracts are able to fully
suppress tumer cell after
4 days.
The antioxidant activity of the extract was found
to be concentration-dependent.
Ref : Characterizing the antioxidant activity of amla(Phyllanthus emblica) extractS. M. Khopde†, K. Indira Priyadarsini†,
H. Mohan†, V. B. Gawandi†,J. G. Satav‡, J. V. Yakhmi†, M. M. Banavaliker**, M. K. Biyani** and J. P. Mittal†,*,#Chemistry
Group, and ‡Radiation Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India**Ajanta
Pharma Ltd, Kandivali, Mumbai 400 067, India#Also with the Jawaharlal Nehru Centre for Advanced Scientific Research,
Bangalore 560 064, India
Antioxidant Activity
The ability of three fractions of E. offcinalis fruit
extract, gallic acid and ascorbic acid for antioxidant activity
Method
: In vitro (DPPH method)
Determined : free radical scavenging by measuring antioxidant
activity (DPPH)
Detection
: UV Spectrophotometer and ESR Spectrometer
Sample
: 3 fraction3 of E. offcinalis fruit extract, gallic acid
and ascorbic acid
Result (antioxidation)
In vitro Antioxidant Activity in Emblica offcinalis
Extract against DPPH and measured by UV Spectrophotometer
Compound
E. officinalis hexane extract
E. officinalis ethyl acetate extract
E.officinalis methanol extract
Ascorbic acid
Gallic acid
IC 50
617.84
2.07
3.82
4.34
2.48
g/ml
g/ml
g/ml
g/m
g/ml
Result (antioxidation)
In vitro Antioxidant Activity in Emblica offcinalis
Extract against DPPH and measured by ESR Spectrometer
Compound
E. officinalis hexane extract
E. officinalis ethyl acetate extract
E.officinalis methanol extract
Ascorbic acid
Gallic acid
IC 50
750.58 g/ml
2.66 g/ml
6.02 g/ml
8.02 g/m
3.67 g/ml
Conclusion (antioxidation)
Three fractions of E. officinalis extract, gallic acid and
ascorbic acid showed antioxidant activity against free radical
(DPPH) both in measure by UV Spectrophotometer and ESR
spectrometer.
The antioxidant activity.
Ethyl acetate ext.  gallic acid  methanol ext.  ascorbic acid  hexane ext.
Evaluation of Emblica officinalis extract cream
Skin lightening
The clinical efficacy of E. officinalis formulations were
evaluated in a placebo-controlled trial using cream base as control.
Method
: In vivo
Subjects
: 20 subjects (20-60 yr.)
Instrument
: Chromameter CR 300
Sample
: 3 % Emblica officinalis extract cream
80
L* parameter
78
76
55°
74
very light
72
70
41°
light
68
66
64
62
28°
medium
60
58
dark
(dark)
56
10°
54
52
ITA
very dark
50
0
2
4
6
8
10
12
14
16
b* parameter
18
20
22
24
26
28
Result
The facial skin conditions were evaluated pre and post
application, determined L- value.
L-value
E. officinalis extract at 3 % w/w showed lightening
effect start at 4 th week.
L*parameter - For clearity;from dark to light (If the L*parameter is high, the skin color is light.)
Beta-glucan
- Beta glucan is polysaccharide derived from the cell wall of bacterial,
fungal, yeasts and cereals.
- Beta glucan has many structure depend on source.
Structure of Beta-glucan
“Beta glucan of yeast (Saccharomyces serevisae) is
mainly of 1,3/1,6-β-D-glucose”
Application
•
•
•
•
•
•
Immunostimulation
Cholesterol reduction
Vaccine adjuvants
Cosmetics for skin repair
Improving food products
Feed supplements
Application
• Anti oxidantion
• Anti inflammation
Application of beta glucan for Immunostimulation
Application of beta glucan for Immunostimulation
Application of beta glucan for Immunostimulation
1200
Cytokine level (pg/ml)
1000
Control
beta-glucan
800
- 6-wk-old BALA/c mice.
600
- Tumor cells were injected into
abdominal wall.
400
- Adding beta-glucan (28.4 mg/kg)
for 21 days.
200
Cytokine
0
IL-2
IFN-g
TNF-a
IL-2
IFN-g
IFN-a
(Interleukin 2 )
(Interferon-γ)
(Interferon –a)
Effect of oral administration b-glucan on cytokines
JANA (2002)
Total cholesterol:HDL cholesterol
Application of beta glucan for cholesterol reduction
10
9.5
9
8.5
8
7.5
7
6.5
6
5.5
5
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
15 obese men ages 20 to 60 with high
cholesterol
Supplement with 7.5 g of beta-glucan in
orange juice twin daily for eight weeks
1
Baseline
7
2
3
8
4
12
Mean changes in ratio of total to HDL-cholesterol concentrations during
supplementation with beta-glucan
Am J Cli Nutr (1999)
Time (wk)
Application of beta glucan for Cosmetics for skin repair
Davis (1992 )and Donzis (1993) were used beta glucan that purified from
yeast cell to cosmetic composition. The products were reduced skin
irritation, lesions, cracks and dryness.
Greene (2001) was applied beta-glucan to cosmetic to for the protection
of skin from damage caused by ultraviolet radiation.
Application of beta glucan for improving food products
New value products to improve health (Wheatcroft et al., 2002)
- Yoghurt
- Fruit juice
- Salad dressing
- Many food products
Improving propertied food products (Thammakiti et al., 2004)
- Thickening
- Water-holding agent
- Emulsifying stabilizer
- Fat replacer