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SKIN WHITENING-A BRIEF REVIEW

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

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and heat it becomes rougher and
thicker. Now a days, wrinkles and skin
darkens are also considered as a
major problem. Skin contains mainly
two types of essential fatty acids,
linoleic acid and arachidonic acid,
having a vital role in regulating barrier
functions. Two types of skin are
present in humans, hairy and non-
hairy. Hair follicles and sebaceous
glands present in hairy skin and non-
hairy skin consists only of sebaceous
glands. Non-hairy skin is present in
palms and soles.
1.1. Epidermis:[3,5,6]
The outer layer of skin works as a
protective sheath against
environmental influences like sunrays,
dust, heat, microbial infection etc.
Epidermis comprises of five separate
layers like;
a. Horny layer (Stratum corneum).
b. Stratum lucidum.
c. Stratum granulosum (Granular
layer).
d. Stratum spinosum (Prickly cell
layer).
e. Stratum germinativum (Basal layer
and dermoepidermal junction).
1.2. Dermis: [3]
Dermis is the middle layer of skin
which gives elasticity, moisture and
firmness to the skin. Blood vessels,
nerve cells, sebaceous glands, sweat
glands are present in dermis layer.
Dermis is having 0.2 to 0.3cm
thickness. It also contains dense
network of structural protein fibers
that is collagen, reticulum and elastin.
1.3. Subcutaneous tissues: [3]
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Subcutaneous layer consists of a sheet
of fat areolar tissue. This type of layer
is quite elastic and having the
presence of large arteries and veins in
the superficial region. It mainly
synthesizes and stores high energy
chemicals.
2. Skin disorders: [7,8]
2.1. Pigmenting disorders- These
are also called as pigmentation
disorders and are due to excessive or
reductive production of melanin. Due
to this disorder, skin becomes
darkens, blotchy (discolored) or
lighter.
2.1.1. Types of Skin Pigmentation:
2.1.1.1. Hypo-pigmentation- It is
caused by lesser amount of melanin.
It can also occur in caucasions. The
common disorders due to hypo-
pigmentation are-
2.1.1.1.1 Vitiligo- It is an auto-
immune disorder which causes white
spots on the skin. Now a day’s 2% of
the population is affected by vitiligo.
Addison’s disease and hyperthyroidism
are common with vitiligo.
2.1.1.1.2 Albinism- It occurs due to
complete absence of pigment. Patient
suffering from this disorder have pale
skin and light yellow eyes. Skin cancer
is common with albinism.
2.1.1.2. Hyper-pigmentation- It is
caused by excessive production of
melanin in the skin. The major factors
which lead to hyper-pigmentation are
excessive amount of sunbathing and
malnutrition. Drugs like antibiotics,
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

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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

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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].

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 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-

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 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]

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 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

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 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

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tumors formation problem. Another
example of anti oxidants like Vitamin
E, Vitamin C, Lentinus edodes,
Pluchea indica inhibits the U.V.
penetration power.
9.3. Vitamins- Vitamins are the vital
compounds which are very beneficial
to the body. Vitamins are used in anti
wrinkle creams, anti aging creams,
skin whitening creams and in the
various products which are very
beneficial to the skin. Vitamin C is
widely used in preparation of
cosmetics as it is having an anti
oxidant property [44]. Vitamin E is
also having an anti oxidant property
and used in cosmetics because of this
free radical scavenging property
[45,46].In human skin it acts as a
predominant lipid soluble anti oxidant
in the human stratum corneum. They
are also used to protect several
components like DNA, proteins,
reactive oxygen species (ROS) which
are caused by UV radiation and by
various air pollutants [47]. Vitamin A
(Retinol) is also used in the cosmetics
and in dietary nutrient required for the
growth of bone and for skin keratosis.
9.4. Emollients-[48] Emollients
having a skin softening function. They
are used to reduce skin dryness, skin
scaling, wrinkles and mild irritant
contact dermatitis. Several emollients
used for skin nourishing are Almond
oil, Caster oil, Coconut oil, Grapeseed
oil, Meadowfoam seed oil, Hemp seed
oil, Jojoba oil, Rose hip oil, Sesame
oil, Squaline, etc.
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9.5. Skin lightening actives-[49]
Skin lightening active agents used to
light the skin color which becomes
darker due to excessive amount of
sunray bathing. There are a lot of skin
lightening formulation present in the
market which may contain anti
oxidant, anti wrinkle, anti aging
properties for the removal of several
skin related problems. Skin lightening
formulations may also contain natural
extracts which influences the
melanization process. Skin lightening
formulations contains different
combinations like Arbutin, Azelaic
acid, Kjoic acid, Resveratrol, Liquorice.
Trichloroacetic acid is used to remove
melanin loaded skin tissues.
10. Testing for Skin care
formulations- Patients suffering with
these types of lupus disorders have to
avoid directly exposure of sunlight or
U.V. light by using various sun
protecting factors like SPF 30, SPF 40
and SPF 50 etc. various anti-malarial
drugs and hydroxychloroquine may be
highly effective in treating various skin
diseases. Skin care formulations to be
used for controlling various skin
related problems like skin darkens,
aging, black spots, acne and many
other problems. Now a day’s both
herbal as well as synthetic skin care
formulations are more generally used.
Herbal cosmetics formulations are
used as they are not having any side
effect to the skin but they are not
easy to use, not easy to handle that’s
why synthetic cosmetics are generally
used as they are easy to use and easy
to handle. Various tests are performed
on skin care formulations such as anti-
oxidant property test, anti- tyrosinase
test etc.
10.1. Anti oxidant property-These
agents are used to inhibit several
problems occurs with skin. This test is
evaluated by several methods like-
10.1.1. DPPH (1, 1-diphenyl-2-
picrylhydrazine) radical scavenging
activity.
10.1.2. ABTS (2, 2-azino-bis (3-
ethylbenzthiazoline-6-sulfonic acid)
scavenging activity.
10.1.3. Nitric acid scavenging
activity.
10.1.4. Hydrogen peroxide (H2O2)
scavenging activity.
10.1.5. Anti oxidant property is also
determined by using FRAP (Ferric
reducing ability of plasma or plant),
Benzie and Strain.
10.1.1. DPPH (1, 1-diphenyl-2-
picrylhydrazine) radical
scavenging activity-[50] DPPH is
commonly used for the anti oxidant
property. DPPH method is generally
used in a 96 well microtitre plate.
When the anti oxidant molecules
incubated, they reacts with DPPH
(which is in purple color) and forms
di-phenyl hydrazine (yellow color).
This conversion of DPPH (purple color)
to di-phenyl hydrazine (yellow color)
measured at 520nm which is used to
measure the scavenging potential of
the plant extracts added to specified
volume of DPPH solution in a
microtitre plate. These plates are
incubated at 25 °C for 10minutes and
absorbance is measured at 520nm.
DPPH acts as a control solvent and
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other solvents used with plant extract
are used as test solvents.
% inhibition of DPPH radical
scavenging activity =
[(Absorbance of control-
Absorbance of test) / (Absorbance
of control)]×100
10.1.2. ABTS(2,2-azino-bis (3-
ethylbenzthiazoline-6-sulfonic
acid) radical cation decoloration
scavenging activity-[51] In this
procedure a specified amount of ABTS
is dissolved in distilled water and also
added potassium persulphate to it.
The whole reaction mixture is allowed
to stand at room temperature for
overnight before use. To various
concentrations of extracts, add
specified volume of DMSO (di-methyl
sulfoxide) and ABTS to make the final
volume. Absorbance is measured at
734nm for 20 minutes.
10.1.3. Nitric acid scavenging
activity-[52,53] Sodium
nitropursside is incubated in
phosphate buffer solution (pH 7.4)
with different concentrations of test
extracts incubated at 25 °C for 5
hours. The incubation plates are
removed and are diluted with Griess
reagent (which is prepared by using
equal volume of 1% sulphanilamide in
2% phosphoric acid and 0.1%
naphthylethylene dihydrochloride in
water). The absorbance is measured
at 546nm.
10.1.4. Hydrogen peroxide (H2O2)
scavenging activity-[54] Phosphate
saline buffer (pH 7.4) is used to
prepare hydrogen peroxide solution.
Various concentrations of few ml of
extracts or standards in
ethanol/methanol are added to
hydrogen peroxide solution.
Absorbance measured at 230nm after
10 minutes. Phosphate buffer without
hydrogen peroxide solution (Blank
solution) is used to compare it.
10.2. Anti tyrosinase activity-[55]
Tyrosinase activity is determined by
using dopachrome method where L-
DOPA is used as a substrate. The
assay method is conducted in a 96-
well microtitre plate and absorbance
of these plates is measured at 475nm
with reference. All samples are
dissolved 50% solution of DMSO. Each
microtitre plate contains sample and
phosphate buffer in the ratio of 1:2
and tyrosinase and L-DOPA in the 1:1
ratio. Results are compared with the
50% DMSO solution in replace of
sample.
% tyrosinase inhibition = (Acontrol –
Asample) / Acontrol ×100.
10.3. Anti- microbial activity-[56-
60]. The anti- microbial activity is
performed by the disc-diffusion test by
using the specified volume of
suspension which is containing few ml
of bacteria spread on the nutrient
medium. Air dried and powdered fruit
materials are extracted in a soxhlet
apparatus by using methanol for 72
hours. Extracts are filtered by using
Whatmann filter paper and
concentrated in vaccum at 40 °C by
the help of rotary evaporator. Filter
paper disc are impregnated with few
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µg of extract/ disc and placed into
agarnutrient. Similarly, negative
controls are prepared by using the
same solvents used to dissolve plant
extracts. The plates are incubated at
35 °C for 24 hours.
10.4. Clinical testing- [61] Clinical
testing is performed for the
determination of results skin of
formulations. Clinical testing of skin
formulations is generally done on
human volunteers or animals who give
the exact consent about the
formulations. This test is carried out
for several weeks or days. The skin
formulation (Cosmaceuticals) is
applied on all the human volunteers or
animals used in study. For evaluation
of skin whitening formulations-
Chromameter is used for the
measurement of skin darkness. Skin
darkness is measured which is taken
before applying the product and again
after few hours. The test carried out
for the result of skin formulations.
10.5. Mushroom tyrosinase
inhibitory assay-[62] Mushroom
tyrosinase inhibitory assay of various
plant extracts are determined by
spectrophotometric methods. In this
method, the plant extracts were
dissolved in DMSO solution at a
constant concentration then diluted it
with different concentrations of DMSO
solution. The sample solution is
diluted with phosphate buffer pH 6.8
in test tubes. After this add one or two
drops of L-Tyrosine solution and finally
added mushroom tyrosinase solution.
Few microlitres of DMSO and Kojic
acid solution are used as blank
reference and positive control. The
initial absorbance is measured at
490nm. Then incubate it for 20
minutes at 37 °C and then the final
absorbance is measured at 490nm.
The IC50 values are determined the
concentration at which the 50%
tyrosinase activity inhibited.
% mushroom tyrosinase inhibition
= [(A2 – A1) – (B2 – B1)] / (A2 –
A1)] × 100
Where, A1 = absorbance of blank
solution at 490nm at 0 minute.
A2 = absorbance of blank solution at
490nm at 20 minutes.
B1 = absorbance of sample solution at
490nm at 0 minute.
B2 = absorbance of sample solution at
490nm at 20 minutes.
10.6. Skin moisture content-[63]
Moisture content for skin mainly
involves the retaining and increasing
water content; reducing the TEWL
[Transepidermal water loss] level,
maintain the skin integrity and
appearance. This study is used to
10.8. Pharmaceutical Stability testing-
10.8.1. General evaluation parameters-
Table 3
S.No. Parameters
1. Total fatty matter
2. Water content
3. Titratable acidity
4. Standard plate count
5. Coliform count
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investigate the effects of base and
formulation on sebum contents of
human skin. The sebum content of
human skin is obtained by the help of
ANOVA test. It is observed that after
application of base an increment occur
in the sebum content may produce an
oily layer on human skin (w/o
emulsion). While reduction in sebum
content occurs shows the presence of
isotretinoin after application which is
effective in reducing the sebaceous
gland size, reducing the sebum
production level by inhibiting the
sebaceous lipid synthesis.
10.7.Transepidermal water loss
(TEWL)–[63] This method is used to
reflect/ show the skin water content.
An increment occurs in TEWL shows
the impairment of water barrier. TEWL
measure the skin irritants and various
allergic patch test reactions. TEWL
measurements affected by skin
surface temperature, ambient air
temperature, ambient air humidity,
anatomical site, sweating and other
various variables.
Limits
22.0-25.0
60.0-70.0
6.0-7.5
Fair
Satisfactory
 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,

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 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.

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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.
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