See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/320930324

SKIN WHITENING-A BRIEF REVIEW

Article · February 2013

CITATIONS READS

0 2,363

3 authors, including:

Aditi Singhal Dharmendra Kumar

Noida Institute of Engineering and Technology Galgotias University
2 PUBLICATIONS   2 CITATIONS    14 PUBLICATIONS   15 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Ancient Natural Remedies View project

air pollution View project

All content following this page was uploaded by Dharmendra Kumar on 08 November 2017.

The user has requested enhancement of the downloaded file.

 SKIN WHITENING- A BRIEF REVIEW

Aditi Singhal*, Dharmendra Kumar, Mayank Bansal

Department of Pharmaceutical Technology,
Meerut Institute of Engineering and Technology,
Baghpat bypass crossing, NH-58, Delhi-Roorkee Highway, Meerut-250005, U.P.,
India.
*singhal28aditi@gmail.com

Abstract:
It is the time for herbal cosmetics as it is the time for looking young and smarter.
They are also being widely used forimproving several skin related problems. Herbal
cosmetics are also called as natural cosmetics. Skin care cosmetics are being used
for several functions like anti-oxidants, anti-inflammatory, anti-microbial, anti-
septic etc. vitamins are also added in skin care cosmetic formulation. Pigmentation
is the reason for various skin related diseases like skin darkens, aging, acne,
wrinkles etc. Anti-oxidant property, anti-microbial activity, skin moisture content
activity, mushroom tyrosinase inhibitory assay etc are various methods for the
evaluation of skin care cosmetics. Natural Cosmaceuticals does not have any side
effects to the skin, easy to handle, easy to store, easy to use and offers various
beneficial effects to the skin.

REFERENCE ID: PHARMATUTOR-ART-1853

Introduction Skin is composed of three layers,

1. Anatomy and Physiology: which are:
[1,2,3,4] 1.1. Cellular epidermis.
Skin is the body’s outer covering layer 1.2. Dermis.
which helps to protect us from heat, 1.3. Subcutaneous Tissues.
light, microbial infection and injury. It
is the body’s largest organ, mainly pH value of skin varies from 4.0 to
regulates body temperature and 5.6. Generally, it is considered as the
stores vitamin D which is essential for smooth layer but because of aging,
human body, fat as well as water. sunrays, dust, microbial infection, cold

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
and heat it becomes rougher and Subcutaneous layer consists of a sheet
thicker. Now a days, wrinkles and skin of fat areolar tissue. This type of layer
darkens are also considered as a is quite elastic and having the
major problem. Skin contains mainly presence of large arteries and veins in
two types of essential fatty acids, the superficial region. It mainly
linoleic acid and arachidonic acid, synthesizes and stores high energy
having a vital role in regulating barrier chemicals.
functions. Two types of skin are
present in humans, hairy and non- 2. Skin disorders: [7,8]
hairy. Hair follicles and sebaceous 2.1. Pigmenting disorders- These
glands present in hairy skin and non- are also called as pigmentation
hairy skin consists only of sebaceous disorders and are due to excessive or
glands. Non-hairy skin is present in reductive production of melanin. Due
palms and soles. to this disorder, skin becomes
darkens, blotchy (discolored) or
1.1. Epidermis:[3,5,6] lighter.
The outer layer of skin works as a
protective sheath against 2.1.1. Types of Skin Pigmentation:
environmental influences like sunrays, 2.1.1.1. Hypo-pigmentation- It is
dust, heat, microbial infection etc. caused by lesser amount of melanin.
Epidermis comprises of five separate It can also occur in caucasions. The
layers like; common disorders due to hypo-
a. Horny layer (Stratum corneum). pigmentation are-
b. Stratum lucidum.
c. Stratum granulosum (Granular 2.1.1.1.1 Vitiligo- It is an auto-
layer). immune disorder which causes white
d. Stratum spinosum (Prickly cell spots on the skin. Now a day’s 2% of
layer). the population is affected by vitiligo.
e. Stratum germinativum (Basal layer Addison’s disease and hyperthyroidism
and dermoepidermal junction). are common with vitiligo.

1.2. Dermis: [3] 2.1.1.1.2 Albinism- It occurs due to

Dermis is the middle layer of skin complete absence of pigment. Patient
which gives elasticity, moisture and suffering from this disorder have pale
firmness to the skin. Blood vessels, skin and light yellow eyes. Skin cancer
nerve cells, sebaceous glands, sweat is common with albinism.
glands are present in dermis layer.
Dermis is having 0.2 to 0.3cm 2.1.1.2. Hyper-pigmentation- It is
thickness. It also contains dense caused by excessive production of
network of structural protein fibers melanin in the skin. The major factors
that is collagen, reticulum and elastin. which lead to hyper-pigmentation are
1.3. Subcutaneous tissues: [3] excessive amount of sunbathing and
malnutrition. Drugs like antibiotics,

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
anti-malarial, anti-arrhythmic can also antioxidant, anti-inflammatory,
lead to hyper-pigmentation. The antibacterial and antiseptic properties.
common hyper-pigmentation disorders Herbal and Synthetic preparations are
are lamellar ichthyosis (fish scale available in market in the form of
disease), lichen simplex and melasma. creams/gels. Massage creams,
vanishing creams and face wash gels
2.2. Sebaceous and Sweat glands are also used to treat various skin
disorders- The common disorders are problems. To protect from U.V.
acne, pimples, blackheads, prickly radiations various sun protecting
heat. This disorder mainly occurs in factors like SPF 15, SPF 30, SPF 40,
neck and large area of skin. SPF 50 are used but SPF 40 and SPF
50 are best suited to protect skin from
2.3. Skin scaling disorders- The U.V. radiations. In herbal cosmetics,
commondisorders are dandruff and natural substances like aloe-vera,
psoriasis. cucumber, turmeric, green tea, rose
etc. are used to treat all types of skin
2.3.1.Psoriasis- It is the formation of problems. Similarly synthetic
scaly red patches on skin. cosmetics like pearl powder, Sebum
REG, BHT (butylated hydroxyl
2.3.2. Dandruff- It can be caused by toluene), Beeswax, Candelilla wax,
microbial infection or immunological Lanolin alcohol, Ozokerite wax,
disorders. It is present on the stratum Ethylene diamine tetra acetate
corneum. (EDTA), Triethanolamine etc are used
to treat skin problem. Skin diseases
2.3.3.Effects of aging on skin- Due are very common symptoms of lupus.
to aging skin become darker, dull, Almost 80% of the lupus suffering
damage and lower melanin level. Acne patients are having several skin
and wrinkle are very common related problems/diseases. Mainly
examples. three types of lupus are as follows;
[10]
3. Skin problems treated by herbal
and Synthetic (man-made) 3.1. Acute cutaneous lupus
cosmetics:[9] erythematosus (ACLE).
Skin problems are generally treated 3.2. Subacute cutaneous lupus
by the help of herbal and synthetic erythematous (SCLE).
cosmetics. Cosmetics are used to
reduce/lower the wrinkles, acne, 3.3. Discoid lupus erythematous
darkness and controls oil secretion. (DLE) or Chronic cutaneous lupus
Various skin problems are occurred by erythematous (CCLE).
the excessive amount of sunrays
bathing, aging and wrinkles. These 3.1.Acute cutaneous lupus
problems are treated by using erythematosus (ACLE) – It is also
cosmetics which are having known as ‘Malar rash’ which involves

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
the upper cheeks, and is also known a. Localized DLE- The lesions are
as ‘Butterfly rash’ which refers unique located on the scalp and face.
butterfly like shape. b. Generalized DLE- Thelesions are
present on any portion of the body.
3.1.1. Characteristics of ACLE-
a. ACLE is having a symmetrical 3.3.2. Characteristics of DLE-
appearance which covers both cheeks a. It may only affect the skin.
and the nose bridge. b. Disk or coin shaped lesions occur.
b. Neck and forehead are also affected c. Painless.
in many cases. d. Usually present on scalp and face.
c. Skin becomes red and swollen
similarly like sunburn. 3.3.3. Causes and long term
3.1.2. Causes and long term effects-
effects- DLE is more photosensitive then SCLE.
ACLE is highly photosensitive. ACLE It may damage hair follicles which
lasts for several weeks or days. It may lead to permanent hair loss. Several
cause skin rashes, but if the rashes environm1111ental factors and
will clear there will be no permanent different types of medications may
effects. used to stimulate the symptoms of
DLE.
3.2.Subacute cutaneous lupus
erythematosus (SCLE) – 4. Melanin Pigment-
3.2.1. Characteristics of SCLE- Melanin is the end-product of various
a. Ring shaped or dish- shaped multiple transformations of L-tyrosine.
patches will occur. Melanin is biosynthesized by;
b. Sun exposure and another form of a. Initiated by the hydroxylation of L-
U.V. light may cause SCLE. phenylalanine to L-tyrosine.
b. Oxidation of L-DOPA to
3.2.2. Causes and long term dopaquinone- is similar for both
effects- eumelanogenesis and
SCLE is also photosensitive; it is pheomelanogenic pathway. [11]
started by U.V. light. Skin becomes Eumelanogenesis involves the
darkening due to this disorder. It can procedure of dopaquinone to
also occur due to side effects of leukodopachrome. Pheomelanogenesis
medication. may also starts with dopaquinone,
conjugate with cysteine or glutathione
3.3.Discoid lupus erythematosus to yield cysteinyldopa and
(DLE) – It is also called as Chronic glutathionyldopa for transformation
cutaneous lupus erythematous into pheomelanin. Melanin pigments
(CCLE). having their common arrangements
linked by carbon-carbon bonds, but
3.3.1. Types of Discoid lupus having different chemical composition,
erythematosus (DLE) - physical as well as chemical

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
properties. Eumelanine is behaving Pheomelanin is highly protein bound
like polyanions, having the capability which indicates that it acts like a
of reversibly bind anions, cations and chromo protein [11,12,13], it can
polyamines. Electron paramagnetic also act as a binding agent for
resonance (EPR) spectrum of chemicals and drugs.[14] Adult
eumelanin yields to slightly female epidermis is less melanized in
asymmetric singlet which produces a comparison to males due to gender
free radical signal. Pheomelanin is the specific effect.[15] Function of
back bone of benzothiazine units melanin in human skin appears by the
which exhibits a yellow to reddish attenuation of U.V. penetration to the
brown color and is soluble in alkali. blood in dermal vessels.[16].

Fig 1: Melanin Formation

5. Melanogenesis Pathway- Melanocytes are usually 7µm in length.Melanin is the

only factor responsible for skin coloration. Melanin is produced in the melanocytes,
where melanocytes stored in the melanosomes after these melanocytes transferred
melanosomes into keratinocytes. Melanocytes are stored in the epidermal-dermal
junction. They extend the intracellular nitric acid (NO) production which initiates
signal transduction cascades for the production of Melanogenesis by the help of
tyrosinase enzyme. [17].

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
 Fig.2: Melanogenesis Pathway

5.1 The process of Melanogenesis boxes, M-boxes as well as tyrosinase

can be understood in following distal element (TDE), five AP-1 and
three steps- Melanogenesis process two AP-2 binding sites are used for
is done by three steps such as; [18] phorbol ester-inducible enhancer-
a. Production of Cysteinyldopa- The binding protein AP-1 (Activator
process occurs with the addition of protein-1), UV responsive elements
cysteine to dopaquinone. (URE), thyroid and retinoic acid like
b. Oxidation of Cysteinyldopa- This responsive elements (TRE and RER),
process gives the pheomelanin. tyrosinase element-1 (TE-1) [20-30].
c. Production of eumelanin- which Transcriptional regulation of
starts after the reduction of Melanogenesis related genes is having
Cysteinyldopa. an important role by the number of
class III POU binding protein Brn-2/N-
5.2 Mechanism of regulation of Oct3[31,32]. Negative
Melanogenesis: [19-35] transcriptional regulation process is
Melanogenesis is regulated by two mediated by brachyury-related
methods which are: transcription factor (TBX 2) [33].
5.2.1. Transcriptional Regulation- 5.2.2. Intracellular signal
Tyrosinase genes generally contains at transduction regulation-
least one major and three minor Intracellular signal transduction
transcription sites.[19] TATA and pathway is used for the positive
CAAT boxes comes under promoter regulation of Melanogenesis with cAMP
region, many potentional used as the critical factor[34,35].
micropthalmia-associated transcription
factor (MITF) binding sites including E-

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
 Fig.3: Intracellular Signal Transduction Pathway

Where, DAG- diacylglycerol ; PKC- applying on skin. Generally herbal

Protein kinase C ; P13-K- cosmetics are derived from botanical
Phosphatidylinositol 3-kinase ; PKA- extracts which are having highly
Protein kinase A ; NO- Nitric acid ; antioxidant property, anti
PKG- Protein kinase G. inflammatory property and rich in
cAMP regulation mechanism of vitamin C and vitamin K. Herbal
Melanogenesis including the activities cosmetics are mostly used by the
of protein kinase A (PKA), then following benefits;
phosphorylates enzymes, ion channels a. Safely to the skin.
and various regulatory proteins. b. Easily available.
Transcriptional activity is regulated by c. Low cost.
activated PKA (Protein kinase A) which d. Having less or no side effects on
involves phosphorylation of cAMP the skin.
responsive element binding protein e. Having high concentration of
(CREB) and CREB binding protein vitamins and minerals which are
(CBP). beneficial for prevention of skin
disease.
6. Herbal cosmetics: [36] f. Almost more than half population
Herbal cosmetics have a great role for totally depends on the herbal
reducing the skin problems which cosmetics as they are having less or
occurs due to several factors. Herbal negligible side effects.
cosmetics have no side effects while

7. List of Some Herbal medicines used for treating Skin diseases- [37,38]

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
 Table 1

Scientific name Family Uses

Bauhinia Variegata Fabaceae Anti-oxidant property

Areca catechu Arecaceae Anti-oxidant property

Aloe Vera Liliaceae Anti-dermatitis

Green tea Theaceae Photo-protective
Ginko bilolsa Ginkogaceae Skin tonic
Allium sativum Alliaceae Anti-oxidant
Dancus carota Apiaceae U.V. protection
Curcuma domestica Tricholomataceae Skin care
Lentinus edodes Zingiberaceae Anti-oxidant property
Muntingia calbura Skin lightening and anti-
Elaeocarpaceae
(Jamaica cherry) oxidant property
Pluchea indica Asteraceae Anti-oxidant property
Glycyrrhiza glabra Leguminosae Skin whitener
Thea viridis Theaceae Anti-oxidant
Crataevea murula Capparidaceae Anti-aging
Rosemarinus officinalis Lamiaceae Anti-aging
Buckwheet seeds Polygonaceae Anti-wrinkle
Psorolia corlifolia Fabaceae Pigmenting agent

8. Synthetic Cosmetics: [39]

Synthetic cosmetics are also used for skin care formulations. Mainly the synthetic
cosmetic preparation having a large amount of surfactants used like Span 20,
Tween 80, Sodium lauryl sulphate, etc. They are less time consuming, easy to use,
easy to carry and easy to store. The various ingredients used in synthetic
preparations are as follows;

Table 2
Synthetic Agents Examples
Anti Aging and Anti Wrinkle Argireline, Pearl powder, Beta
agents Glucan, Coenzyme Q10, Glycan

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
 Booster peptide, Ceramide complex
Soothing and Regenerating Allantoin, Aloe-Vera gel, Colloidal
agents Oatmeal
Oily Skin Regulators Sebum REG
Alpha- Arbutin, Skin white BLE, Kojic
Skin Whitening agents acid, Skin white MSH, Glycolic acid,
Aleosin, Hydroquinone, Salicylic acid
Emollients Squaline
Glycerin, Sodium PCA, Caprylyl
Gylcol EHG, Algae extract and
Humectants
Hyaluronate gel, Propylne glycol,
Hyaluronic acid, Sorbital, Urea
Alkyl sulphonate, Coco betaine, Coco
Surfactants glucose, Sulfosuccinate, Polyglucose,
Polyquaternium-87
Ethylene diamine tetra acetate
Emulsion stabilizers
(EDTA), Triethanolamine
BHT(Butylated hydroxyl toulene),
Anti-oxidants
propyl gallate, Vitamin E
Lanolin alcohol, , Cetyl alcohol,
Thickeners
Carbomer, Acrylate copolymer

9. Functional Active Agents in Skin aging cream mainly acts to slow or

care formulations: reduce the signs of aging.

9.1Anti Aging agents-[40] Stress is 9.2Anti Oxidant agents-[41-43]

the major factor for aging. Other These agents are used to inhibit the
factors which cause aging are excess several problems of skin. Reactive
amount of calories present in the Oxygen Species (ROS) is the main
body, excessive insulin amount, factor responsible for several skin
reduced anti oxidant level, etc. Due to problems. Anti oxidants are used to
aging the skin becomes less elastic, inhibit the ROS production. Anti
becomes drier, and wrinkles appears oxidants works at different levels in
in huge amount. Skin aging problem oxidative process such as;
may breakdown the DNA, Collagen, a. Free radical scavenging.
Elastin, Hyaluronic acid and all other b. Lipid peroxyl radicals scavenging.
molecules present in the skin. c. Metal ion binding occurs.
Melanotropin activity, Tyrosinase d. Removing oxidatively damaged
activity becomes larger due to skin biomolecules.
aging problem. Reduction of Vitamin C Ascorbic acid is the main and effective
and Vitamin E are the main factors example of anti oxidant used to
that causes skin aging problem.Anti- control sun burn, cell formation and

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
tumors formation problem. Another 9.5. Skin lightening actives-[49]
example of anti oxidants like Vitamin Skin lightening active agents used to
E, Vitamin C, Lentinus edodes, light the skin color which becomes
Pluchea indica inhibits the U.V. darker due to excessive amount of
penetration power. sunray bathing. There are a lot of skin
lightening formulation present in the
9.3. Vitamins- Vitamins are the vital market which may contain anti
compounds which are very beneficial oxidant, anti wrinkle, anti aging
to the body. Vitamins are used in anti properties for the removal of several
wrinkle creams, anti aging creams, skin related problems. Skin lightening
skin whitening creams and in the formulations may also contain natural
various products which are very extracts which influences the
beneficial to the skin. Vitamin C is melanization process. Skin lightening
widely used in preparation of formulations contains different
cosmetics as it is having an anti combinations like Arbutin, Azelaic
oxidant property [44]. Vitamin E is acid, Kjoic acid, Resveratrol, Liquorice.
also having an anti oxidant property Trichloroacetic acid is used to remove
and used in cosmetics because of this melanin loaded skin tissues.
free radical scavenging property
[45,46].In human skin it acts as a 10. Testing for Skin care
predominant lipid soluble anti oxidant formulations- Patients suffering with
in the human stratum corneum. They these types of lupus disorders have to
are also used to protect several avoid directly exposure of sunlight or
components like DNA, proteins, U.V. light by using various sun
reactive oxygen species (ROS) which protecting factors like SPF 30, SPF 40
are caused by UV radiation and by and SPF 50 etc. various anti-malarial
various air pollutants [47]. Vitamin A drugs and hydroxychloroquine may be
(Retinol) is also used in the cosmetics highly effective in treating various skin
and in dietary nutrient required for the diseases. Skin care formulations to be
growth of bone and for skin keratosis. used for controlling various skin
related problems like skin darkens,
9.4. Emollients-[48] Emollients aging, black spots, acne and many
having a skin softening function. They other problems. Now a day’s both
are used to reduce skin dryness, skin herbal as well as synthetic skin care
scaling, wrinkles and mild irritant formulations are more generally used.
contact dermatitis. Several emollients Herbal cosmetics formulations are
used for skin nourishing are Almond used as they are not having any side
oil, Caster oil, Coconut oil, Grapeseed effect to the skin but they are not
oil, Meadowfoam seed oil, Hemp seed easy to use, not easy to handle that’s
oil, Jojoba oil, Rose hip oil, Sesame why synthetic cosmetics are generally
oil, Squaline, etc. used as they are easy to use and easy
to handle. Various tests are performed
on skin care formulations such as anti-

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
oxidant property test, anti- tyrosinase other solvents used with plant extract
test etc. are used as test solvents.

10.1. Anti oxidant property-These % inhibition of DPPH radical

agents are used to inhibit several scavenging activity =
problems occurs with skin. This test is [(Absorbance of control-
evaluated by several methods like- Absorbance of test) / (Absorbance
10.1.1. DPPH (1, 1-diphenyl-2- of control)]×100
picrylhydrazine) radical scavenging
activity. 10.1.2. ABTS(2,2-azino-bis (3-
10.1.2. ABTS (2, 2-azino-bis (3- ethylbenzthiazoline-6-sulfonic
ethylbenzthiazoline-6-sulfonic acid) acid) radical cation decoloration
scavenging activity. scavenging activity-[51] In this
10.1.3. Nitric acid scavenging procedure a specified amount of ABTS
activity. is dissolved in distilled water and also
10.1.4. Hydrogen peroxide (H2O2) added potassium persulphate to it.
scavenging activity. The whole reaction mixture is allowed
10.1.5. Anti oxidant property is also to stand at room temperature for
determined by using FRAP (Ferric overnight before use. To various
reducing ability of plasma or plant), concentrations of extracts, add
Benzie and Strain. specified volume of DMSO (di-methyl
sulfoxide) and ABTS to make the final
10.1.1. DPPH (1, 1-diphenyl-2- volume. Absorbance is measured at
picrylhydrazine) radical 734nm for 20 minutes.
scavenging activity-[50] DPPH is
commonly used for the anti oxidant 10.1.3. Nitric acid scavenging
property. DPPH method is generally activity-[52,53] Sodium
used in a 96 well microtitre plate. nitropursside is incubated in
When the anti oxidant molecules phosphate buffer solution (pH 7.4)
incubated, they reacts with DPPH with different concentrations of test
(which is in purple color) and forms extracts incubated at 25 °C for 5
di-phenyl hydrazine (yellow color). hours. The incubation plates are
This conversion of DPPH (purple color) removed and are diluted with Griess
to di-phenyl hydrazine (yellow color) reagent (which is prepared by using
measured at 520nm which is used to equal volume of 1% sulphanilamide in
measure the scavenging potential of 2% phosphoric acid and 0.1%
the plant extracts added to specified naphthylethylene dihydrochloride in
volume of DPPH solution in a water). The absorbance is measured
microtitre plate. These plates are at 546nm.
incubated at 25 °C for 10minutes and
absorbance is measured at 520nm. 10.1.4. Hydrogen peroxide (H2O2)
DPPH acts as a control solvent and scavenging activity-[54] Phosphate
saline buffer (pH 7.4) is used to

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
prepare hydrogen peroxide solution. µg of extract/ disc and placed into
Various concentrations of few ml of agarnutrient. Similarly, negative
extracts or standards in controls are prepared by using the
ethanol/methanol are added to same solvents used to dissolve plant
hydrogen peroxide solution. extracts. The plates are incubated at
Absorbance measured at 230nm after 35 °C for 24 hours.
10 minutes. Phosphate buffer without
hydrogen peroxide solution (Blank 10.4. Clinical testing- [61] Clinical
solution) is used to compare it. testing is performed for the
determination of results skin of
10.2. Anti tyrosinase activity-[55] formulations. Clinical testing of skin
Tyrosinase activity is determined by formulations is generally done on
using dopachrome method where L- human volunteers or animals who give
DOPA is used as a substrate. The the exact consent about the
assay method is conducted in a 96- formulations. This test is carried out
well microtitre plate and absorbance for several weeks or days. The skin
of these plates is measured at 475nm formulation (Cosmaceuticals) is
with reference. All samples are applied on all the human volunteers or
dissolved 50% solution of DMSO. Each animals used in study. For evaluation
microtitre plate contains sample and of skin whitening formulations-
phosphate buffer in the ratio of 1:2 Chromameter is used for the
and tyrosinase and L-DOPA in the 1:1 measurement of skin darkness. Skin
ratio. Results are compared with the darkness is measured which is taken
50% DMSO solution in replace of before applying the product and again
sample. after few hours. The test carried out
for the result of skin formulations.
% tyrosinase inhibition = (Acontrol –
Asample) / Acontrol ×100. 10.5. Mushroom tyrosinase
inhibitory assay-[62] Mushroom
10.3. Anti- microbial activity-[56- tyrosinase inhibitory assay of various
60]. The anti- microbial activity is plant extracts are determined by
performed by the disc-diffusion test by spectrophotometric methods. In this
using the specified volume of method, the plant extracts were
suspension which is containing few ml dissolved in DMSO solution at a
of bacteria spread on the nutrient constant concentration then diluted it
medium. Air dried and powdered fruit with different concentrations of DMSO
materials are extracted in a soxhlet solution. The sample solution is
apparatus by using methanol for 72 diluted with phosphate buffer pH 6.8
hours. Extracts are filtered by using in test tubes. After this add one or two
Whatmann filter paper and drops of L-Tyrosine solution and finally
concentrated in vaccum at 40 °C by added mushroom tyrosinase solution.
the help of rotary evaporator. Filter Few microlitres of DMSO and Kojic
paper disc are impregnated with few acid solution are used as blank

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
reference and positive control. The investigate the effects of base and
initial absorbance is measured at formulation on sebum contents of
490nm. Then incubate it for 20 human skin. The sebum content of
minutes at 37 °C and then the final human skin is obtained by the help of
absorbance is measured at 490nm. ANOVA test. It is observed that after
The IC50 values are determined the application of base an increment occur
concentration at which the 50% in the sebum content may produce an
tyrosinase activity inhibited. oily layer on human skin (w/o
emulsion). While reduction in sebum
% mushroom tyrosinase inhibition content occurs shows the presence of
= [(A2 – A1) – (B2 – B1)] / (A2 – isotretinoin after application which is
A1)] × 100 effective in reducing the sebaceous
gland size, reducing the sebum
Where, A1 = absorbance of blank production level by inhibiting the
solution at 490nm at 0 minute. sebaceous lipid synthesis.
A2 = absorbance of blank solution at
490nm at 20 minutes. 10.7.Transepidermal water loss
B1 = absorbance of sample solution at (TEWL)–[63] This method is used to
490nm at 0 minute. reflect/ show the skin water content.
B2 = absorbance of sample solution at An increment occurs in TEWL shows
490nm at 20 minutes. the impairment of water barrier. TEWL
measure the skin irritants and various
10.6. Skin moisture content-[63] allergic patch test reactions. TEWL
Moisture content for skin mainly measurements affected by skin
involves the retaining and increasing surface temperature, ambient air
water content; reducing the TEWL temperature, ambient air humidity,
[Transepidermal water loss] level, anatomical site, sweating and other
maintain the skin integrity and various variables.
appearance. This study is used to

10.8. Pharmaceutical Stability testing-

10.8.1. General evaluation parameters-

Table 3
S.No. Parameters Limits
1. Total fatty matter 22.0-25.0
2. Water content 60.0-70.0
3. Titratable acidity 6.0-7.5
4. Standard plate count Fair
5. Coliform count Satisfactory

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
 10.8.2. Organoleptic evaluation-
Table 4
S.No. Specifications Limits
1. State Semisolid
2. Odour Characteristic
3. Colour White
4. Spreadbility Easily
5. Oily / tacky film No

10.8.3. Physical evaluation-

Table 5
S.No. Specification Limits
1. Specific gravity 1.00-1.50g /ml
2. Reference index Not more than 2
3. Clarity Clear
4. Viscosity 4000 cp
5. pH 6.0-7.0

10.8.4. Microbial evaluation-

Table 6
S.No. Microbial load Limits
1. Total microbial count Not more than 100
Limit tests- E.coli, S.aures,
2. No characteristic colonies
Solmonella

Marketed Skin care products-

Table 7
Company
S.No. Product Ingredients Reference
name
Kojic acid, lemon
Premium
1. Skin bright extract, [64]
naturals
Arbutin
Arbutin, lumi
Skin
2. skin(diacetyl bolidine), Revitol [65]
brightener
antioxidants,

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
 Vit.A, C and K,
whitener agent
Niacinamide, alpha
arbutin, Kojic acid,
3. Lucederm Sisquoc [66]
lemon, licorice,
Mulberry, bearberry
Kojic acid,
Niacinamide, alpha
arbutin, Mulberry, Civant skin
4. Meladerm [66]
bearberry, licorice, care
giga white, lemon
juice
Sophora angustifolia
5. Synerlight root, LIBiol [65]
actinidia chinensis fruit
Dimethylmethoxy
6. Chromabright Lipotec [65]
chromanyl palmitate

Future development- Skin care Ultra structure of Human Skin.

products are now days most widely London: Churchill, (1971).
used. Mainly herbal cosmetics are 3.Fuchs, E., Raghava, S. Getting
generally used as they does not have under the skin of dermal
side effect to the skin. In coming next morphogenesis. Nat Rev Genet 7,
years it will be expected that there are 199-209 (2002).
a lot of chances for the future 4.Skin disease or dermatology-skin
development of herbal cosmetics. pigmentation disorders/Medindia
Various herbal and synthetic products medindia.net/patients/patient
are to be used for improving various info/skin-disease-skin pigmentation
skin related diseases related to skin disorders.htm#ixzzliXRg0Q00.
darken, acne, wrinkle, sun burn etc. 5.Skin Disease or Dermatology - Skin
New development will give a better pigmentation disorders | Medindia
opportunity in the treatment of skin medindia.net/patients/patientinfo/skin
related diseases. disease-skin-pigmentation-
disorders.htm#ixzz1uXSazgYv.
References: 6.Sharma, R.K., Sambita, C.
1.Toma, JG., Akhaven, M., Ferandes, Bhagwandas Chowkambha Sanskrit
and KJ et al. Isolation of multipoint Series office, Varanasi, 51-56 (1988).
adult stem cells from the dermis of 7.The Ayurvedic Formulary of India,
mammalian skin, Nat Cell Bio 3, 778- Part-I, Govt. of India, Ministry of
784 (2001). Health and Family planning.
2.Breathnach, AS. An Atlas of the Department of Health, 103-119

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
(2003). Pathophysiology. New York: Oxford
8.Ashawat, M.S., Banchhor and M., Univ. Press (1998).
Saraf, S. Herbal cosmetics: “Trends in 17.Bertolotto, C., Abbe, P., Hemesath,
Skin Care Formulation”. Phcog Rev. 3 TJ., Bille, K., Fisher, DE., Ortonne, JP.
(5), 82-89 (2009). and Ballotti, R. Microphthalmia gene
9.11.Jayaprakasha, G.K., et al. product as a signal transducer in
Antioxidant activities of Flavidin in cAMP-induced differentiation of
Different Invitro Model Systems. Bio melanocytes. J Cell Biol 142, 827–835
organic and Med Chem. 12, 5141- (1998).
5146 (2004). 18.Englaro, W., Bahadoran, P.,
10.Prota, G. The chemistry of Bertolotto, C., Busca, R., Derijard, B.,
melanins and melanogenesis. Fortsch Livolsi, A., Peyron, JF., Ortonne, JP.
Chem Organ Natur 64, 93–148 and Ballotti, R. Tumor necrosis factor
(1995). alpha-mediated inhibition of
11.13.Boyan, BD., Bonewald, LF., melanogenesis is dependent on
Sylvia , VL., Nemere, I., Larsson, D., nuclear factor kappa B activation.
Norman, AW., Rosser, J. and Dean, Oncogene 18, 1553–1559 (1999).
DD., Schwartz, Z. Evidence for distinct 19.Ganss, R., Montoliu, L., Monaghan,
membrane receptors for 1 alpha,25- AP. and Schutz, G. A cell specific
(OH)(2)D(3) and 24R,25-(OH)(2)D(3) enhancer far upstream of the mouse
in osteoblasts. Steroids 67, 235–246 tyrosinase gene confers high level and
(2002). copy number-related expression in
12.Robins AH. Biological Perspectives transgenic mice. EMBO J 13, 3083–
on Human Pigmentation. Cambridge, 3093 (1994).
UK: Cambridge Univ. Press (1991). 20.Ganss, R., Schmidt, A., Schutz, G.
13.Lindquist, NG. Accumulation of and Beermann, F. Analysis of the
drugs on melanin. Acta Radiol Diagn mouse tyrosinase promoter in vitro
(Stockh) 325, 1–92 (1973). and in vivo. Pigment Cell Res 7, 275–
14.Romero, GC., Aberdam, E., Biagoli, 278 (1994).
N., Massabni, W., Ortonne, J. P. and 21.Ganss, R., Schutz, G. and
Ballotti, R. 1996. Ultraviolet B Beermann, F. The mouse tyrosinase
radiation acts through the nitric oxide gene. Promoter modulation by positive
and cGMP signal transduction pathway and negative regulatory elements. J
to stimulate melanogenesis in human Biol Chem 269, 29808–29816 (1994).
melanocytes. J. Biol. Chem. 271, 22.Kikuchi, H., Miura, H., Yamamoto,
28052-28056. H., Takeuchi, T., Dei, T. and
15.Shosuke, ito, IFPCS Presidential Watanabe, M. Characteristic
Lecture, A Chemist’s View of sequences in the upstream region of
Melanogenesis. pigment cell res 16, the human tyrosinase gene. Biochim
230-236 (2003). Biophys Acta 1009, 283–286 (1989).
16.Nordlund, JJ., Boissy, RE., Hearing, 23.Kwon, BS. Pigmentation genes: the
VJ., King, RA. and Ortonne, JP. The tyrosinase gene family and the pmel
Pigmentary System. Physiology and 17 gene family. J Invest Dermatol

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
100, 134S–140S (1993). Mol Cell Biol 18, 5099–5108 (1998).
24.Kwon, HY., Bultman, SJ., Loffler, 31.Lerner, AB., Moellmann, G., Varga,
C., Chen, WJ., Furdon, PJ., Powell, VL., Halaban, R. and Pawelek, J.
JG., Usala, AL., Wilkison, W., Action of melanocyte-stimulating
Hansmann, I. and Woychik, RP. hormone on pigment cells. Cold Spring
Molecular structure and chromosomal Harbor Conf Cell Prolif 6, 187–197
mapping of the human homolog of the (1979).
agouti gene. Proc Natl Acad Sci USA 32.Pawelek, J., Wong, G., Sansone, M.
91, 9760–9764 (1994). and Morowitz, J. Molecular biology of
25.Montoliu, L., Umland, T. and pigment cells. Molecular controls in
Schutz, G. A locus control region at 12 mammalian pigmentation. Yale J Biol
kb of the tyrosinase gene. EMBO J 15, Med 46, 430–443 (1973).
6026–6034 (1996). 33. lupuscanada.org.
26.Shibata, K., Muraosa, Y., Tomita, 34.Balakrishnan, K.P. Tyrosinase
Y., Tagami, H. and Shibahara, S. inhibition and anti oxidant properties
Identification of a cis-acting element of Muntingia Calabura extracts, in-
that enhances the pigment cell- vitro studies. Int J Pharma and Bio
specific expression of the human Sci. 2(1).
tyrosinase gene. J Biol Chem 267, 35.Nam, KY., Nuhu, A., Seong, JL.,
20584–20588 (1992). Rom, KL. and Soo, LT. Detection of
27.Shibata, K., Takeda,K., Tomita,Y., Phenolic compounds concentration and
Tagami, H. and Shibahara, S. evaluation of Anti oxidant and Anti
Downstream region of the human tyrosinase activities of various extract
tyrosinase-related protein gene from the fruiting bodies of Lentinum
enhances its promoter activity. edodes. World Applied Sci J 12(10),
Biochem Biophys Res Commun 184, 1851-1859 (2011).
568–575 (1992). 36.Kuno, N., Matsumoto, M. Skin
28.Eisen, T., Easty, DJ., Bennett, DC. beautifying agent, Anti Aging agent for
and Goding, CR. The POU domain Skin Whitening agent and external
transcription factor Brn-2, elevated agent for the Skin. US Patent 6682763
expression in malignant melanoma (2004).
and regulation of melanocyte-specific 37.Smith, W.P. Barrier disruption
gene expression. Oncogene 11, 2157– treatments for structurally
2164 (1995). deteriorated skin. US Patent 5720963
29.Thony, B., Auerbach, G. and Blau, (1998).
N., Tetrahydrobiopterin biosynthesis, 38.Lo, W.B., Black, H.S. Inhibition of
regeneration and functions. Biochem J Carcinogen Formation in skin irritated
347, 1–16 (2000). with ultraviolet light. Nature 246, 489-
30.Carreira, S., Dexter, TJ., Yavuzer, 491 (1973).
U., Easty, DJ. and Goding, CR. 39.Halliwell, B., Gutteridge, JMC. Free
Brachyury-related transcription factor radicals in biology and medicine. 4th
Tbx2 and repression of the edition, New York Oxford University
melanocyte-specific TRP-1 promoter. Press (2007).

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
40.Black, H.S., Lowe, N.J. and 49..Munir, O., et al. Determination of
Hensby, C.N. Role of reactive oxygen in vitro antioxidant activity of fennel
species in inflammatory process. seed extract. Lebensm-Wiss.U-Technol
Pharmacology and the Skin. 2, 1-20 36, 263-271 (2003).
(1989). 50.Mruthunjaya, K., Hukkeri, V.I In
41.Dunham, W.B., Zuckerkandl, E., vitro antioxidant and free radical
Reynolds, R., Willoughby, R., scavenging potential of Parkinsonia
Marcuson, R., R, Barth, R. and aculeate Linn. Pharmacognosy
Pauling, L. Effects of intake of L- Magazine 4, 42-51 (2008).
ascorbic acid on the incidence of 51.Badami, S., et al. In vitro activity
dermal neoplasms induced in mice of various extracts of Aristolochia
ultraviolet light.Proc. Natl. Acad. Sci. bracteolate leaves. Oriental Pharmacy
79, 7532-7536 (1982). and Exp Med. 5, 316-321 (2005).
42.Shindo, Y., Witt, E., Han, D., 52.Chan, E.W.C., Lim, Y.Y., Wong,
Epstein, W. and Packer, L. Enzymic L.F., Lianto, F.S., Wong, S.K., Lim,
and non enzymic antioxidants in K.K., Joe, C.E. and Lim, T.Y.
epidermis and dermis of human skin. Antioxidant and tyrosinase inhibition
J.Invest. Dermatol.102, 122-124 properties of leaves and rhizomes of
(1994). ginger species. Food Chemistry 109,
43.Sies, H., Stahl, W., Sundquist, A.R. 477–483 (2008).
Antioxidant functions of vitamins. Ann. 53.Lin, J., Opoku, A.R., Geheeb, KM.,
NY Acad. Sci. 669, 7-20 (1992). Hthings, A.D., Terblanche, S.E., Jager,
44.Traber, M.G., Sies, H. Vitamin E in A.K. and Van, JS. Preliminary
Human Demand and Delivery. Ann. screening of some traditional zulu
Re. Nutri. 16, 321-347 (1996). medicinal plants for anti-inflammatory
45.Kurata, Y. New Raw materials and and anti-microbial activities. J
technologies in cosmetics: Properties Ethnopharmacology 68, 267-274
and applications of plant extract (1999).
complexes. Fragr. J. 22, 49-53 54.Kim, J., Marshall, M.R. and Wie, C.
(1994). Antibacterial activity of some essential
46.Lee, K.K., Choi, J.J., Park, E.J. and oil components against five food borne
Choi, J.D. Antielastase and pathogens. J Agriculture and Food
Antihyaluronidase of phenolic Chem, 43, 2839-2845 (1995).
substance from Areca Catechu as a 55.Djipa, C.D., Delmee, M. and
new Antiaging agent. Int. J. Cosmet. Quetin-lelcreqc, J. Antimicrobial
Sci. 23, 341-346 (2001). activity of bark extracts of Syzygium
47.49.Ito, S. The IFPCS presidential jambos L. J Ethnopharmacology 71,
lecture: a chemist’s view of 307-313 (2000).
Melanogenesis. Pigment Cell Res 16, 56.Swanson, K.M.S., Busta, F.F.,
230–236 (2003). Peterson, E.H. and Johanson, M.G.
48.Prota, G. Melanins and Colony count methods. In:
Melanogenesis. New York. Academic Vanderzant, C., Splittstoesser, D.F.
(1992). (eds.) Compendium of methods for

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.
 microbiological examination of food. from the Roots of Morus nigra. A
3rd edition. American Public Health Structure-Activity Relationship Study.
Asso, Washington, 75-95 (1992). J. Agric. Food Chem 58, 5368–5373
57.Ozturk, S., Ercisli, S. Antibacterial (2010).
activity and chemical constitutions of 60.Naveed, A., Mehmood, A., Khan,
Ziziphora clinopodioides. Food Control B.A., Mahmood, T., Muhammad, H.,
18(5), 535-540 (2007). Khan, S. and Saeed, T. Exploring
58.Majmudar, G., Jacob, G., Laboy, Y. cucumber extract for skin
and Fisher, L., Mary Kay Holding rejuvenation. African J Bio 10(7),
Corporation, Dallas, TX 75247. An in 1206-1216 (2011).
vitro method for screening skin- 61. whiter skin.com/
whitening products. J. Cosmet. Sci. 62. naturalskincareformula.com/
49, 361-367. 63. whiterskin.com/
59.Zheng, ZP., Cheng, KW., Zhu, Q., 64. whiterskin.com/studies/cosmo.pdf
Wang, XC, Lin, ZX. and Wang, M. 65. gattefossecanada.ca/
Tyrosinase Inhibitory Constituents 66. lipotec.com;patent

FIND THIS ARTICLE ONLINE: http://www.pharmatutor.org/articles/skin-

whitening-a-brief-review

Article by PharmaTutor
www.pharmatutor.org
Do not copy, All rights are reserved.

View publication stats