GEETHANJALI COLLEGE OF

PHARMACY
METHODS OF ANALYSIS TO DETERMINE THE
QUALITY OF COSMECTICS IN THE FINISHED
FORMS

Presented By

Sonia Chowdhury

Mpharm 1st year (Pharmaceutics)

Introduction :
Word Cosmetic Is Originated From Greek Word “kosmeticos” means
adorn and preparation

Definition:
It is external preparation meant for applying on external parts of the body i.e. nails,skin,
hairs for coloring,covering, softening, cleaning, Nourishing ,waving ,setting, preservation,
removal and protection.

Other way of defining cosmetic is“it is item intended to be rubbed,poured, sprinkled ,

introduced in to,otherwise applied to the human body or part thereof for cleaning,
beautifying, promoting attractivenessor altering the appearance”.
 Skin Care Hair Care

Analysis
Baby Care of Personal
Cosmetics Hygiene

Dental Care
 Analysis of cosmetics includes
1) Physical analysis of cosmetic
2) Microbiological analysis of
cosmetics
3) Chemical analysis of cosmetics
1) Physical Analysis Of Cosmetics :
Skin Care Products

Skin Cream Hand Sunscreen Face Special

a)Make up Cream or Powders Cream
-Vanishing Body lotion a)Anti Acne
-Foundation a)Hand Cleanser b)Anti Ageing
b)Cold Cream b)Hand lotion c)All Medicated
c)Moisturizer c)Body lotion Creams
d)Night Cream d)Shower gel
e)Protective cream
f)Cleansing cream
 Evaluation of Skin Cosmetics :
General Sensitivity test :
For Primary potential irritants Draize‟s test use. In this test Albino rabbits are
clipped and substance to be tested is applied to ,
- Intact skin,
- Abrased Skin
- Lightly scarified skin
All of them are covered with a patch for 24 hours and changes are
assessed.BIS 411:1997 suggests that if there is no reaction in any of the animals,
the same testshould be performed on 10 Humans volunteers applying the substances
on the skin offorearm.

As per IS, this test can be carried out on 10 lab.

Animals either guinea pig or rabbits. Cosmetic is applied on 2 cm sq.
( for better exposure, area for test is shaved).
PATCH TEST :
It has two purposes

a) Diagnostic : To discover
whether the cosmetic used
has caused dermatitis.

b) Prophetic : To assess
whether a new cosmetic
should be placed on market
or not
 GENERAL PROCEDURE :
 0.1 to 0.3 gm of cosmetic to be tested is applied on a piece of cotton
fabric, size 2-3 cm sq. and apply this to skin of arms, thigh or back.

 This patch is covered with a patch of cellophane abt 5 cm sq. and sealed
with adhesive
plaster about 40 cm sq.

 Allow to remain on skin for 24-72 hours.o Sites of patched are examined
after
30 minutes of removal of patch by an experienced dermatologist.
(observation can be done earlier but before NLT 15 min)
 Observations and their Grade :
If no reaction, subjects should be observed
for 3 to 5 days to ascertain any late reaction.
Its advisable to find out whether material
causes photo sensitization.

If there is reaction, further test are

necessary to find out which
ingredient is responsible for
reaction. If no reaction……same
reapplied to same place or fresh
patch may be applied…….its
continued till, -Either a reaction is
produced under one or more
patches -Or investigator is
confirmed that no reaction will
occur.
 a) Open Patch Test : - cosmetics with higher % of potential irritants like hair dyes,
shampoos, hair tonics, patches should not be sealed
.- Performed on sensitive part of skin, bend of elbow, skin behind ears.
- Applied on 1 sq m.- Control standard patches- Inspection after 24 hours.

b) Prophetic Patch test :-Before it, investigator should perform test on him self to
ascertain the material is not primary irritant.
- It performed on 10 humans,
- If favorable results, for full scale , 200 normal subjects are used.
Subjects are observed for a days. After 7-10 days, reapplied on, who didn‟t
show reaction
- If no reaction……or one or two of 200…….New product can be
placed on trial sale. BIS 4011:1982….Method for dermatological test
For cosmetics recommends that initial test should be done on 50volunteers.
- If any shows positive reaction, number of volunteers is increased to 200
2) Repeated Insult Test : - The prophetic patch test has certain shortcomings,
to name

- Quick absorption of potential skin irritant through skin

- Rapid evaporation of volatile skin irritant from patch
- small amount of cosmetic in patch in comparison to large amount in actual use
- Small area of skin used.
- Short exposure of skin to cosmetic than the exposure actual in use.

So, Draize described …Repeated Insult Test involves applying of the same
concentration as is found in finished formulation incorporated in a bland base.
Bland base is selected acc. To nature & solubility of the test substance.
3) Photo patch Test :

- Certain substances are not harmful by themselves but they

become harmful whenexposed to sunlight..

- Substances that absorb lightin between 300-308 nm have potential of phototoxicity..

- So when a substance is considered phototoxic, this test may be performed.

- Same as standard patch test, duplicate patches are applied and after 24 hrs one
of the patches in pair is exposed to sunlight for 30 minutes & covered again.
4) Provocative Patch Test :

Test for sensitizing potential With first exposure, generally weak sensitizing agent
don‟t sensitize a person but repeated exposure may make a person hypersensitive
This can be indicated by this test.

• Its performed as standard procedure, 10-15 application on alternate days on

same spot of skin in 10 volunteers.

• Indian standards says in case of unknown chemicals, test should be

first performed on animals like guinea pig or rabbits.
SUNSCREEN PRODUCTS :

Post sweating SPF determined

1. 1000-4000 A Ultraviolet zone.
- 2200-3200 A Therapeutic UV Zone i.e.Vitamin D & anti rachitic vitamin are found.
- 2500-3020 A close to rays that cause sunburn
- 2800-3100 A cause sunburn and are screen out with sunscreen products.
(UVB) so the longer rays of 3000-3200 cause reddening of skin or erythema,
Tan producing rays.

• Sunscreen may be Preventive i.e . sunburnprevention by shading

of body surface simulatory &therapeutic i.e. use of chemicals that screen out certain
rays of sun
SUNSCREEN PRODUCTS :

Post sweating SPF determined

1. 1000-4000 A Ultraviolet zone.
- 2200-3200 A Therapeutic UV Zone i.e.Vitamin D & anti rachitic vitamin are found.
- 2500-3020 A close to rays that cause sunburn
- 2800-3100 A cause sunburn and are screen out with sunscreen products.
(UVB) so the longer rays of 3000-3200 cause reddening of skin or erythema,
Tan producing rays.

• Sunscreen may be Preventive i.e . sunburnprevention by shading

of body surface simulatory &therapeutic i.e. use of chemicals that screen out certain
rays of sun
 FACE POWDERS :
Face powder may be either of loose fine powder or compact .
Evaluation:1) Fineness of Powder -Sieving method ,
-Microscopic Method
-Air separation technique.
As per IS 3959-2004,Residue on 75 μ sieve should be NMT 2 % & 0n 150 μNMT 0.5 %.
2) Apparent Density
3) Shade & Uniformity of shade
- Comparison with standard shade kept for this purpose.
- Commonly std and sample both are placed between two glassplates and compared
Observed in natural light.
4) Odor - No physical measure for odor.
5) Pressure applied on compact powder : by penetro meter,
6) Breaking point. Cake is dropped on wooden (8-10 in) or thick rubber mat (6 feet),
5) Matter insooluble in water : Boil 1 gm. with 200ml,filter, dry residue & find out.
6) Moisture & volatile matter : By drying powder at 105 C to constant weight.
7) pH of aqueous solution : By making suspension in water of 10 % or filtrate 19 may be
used.
The pay-offcharacter, i.e.adhesion with the puff of
compact or pressed powder should betested on
the skin

Pay off 20 Dispersion Analyzer- Instruments

 SPECIAL CREAMS :

Includes

a) Anti Acne : Acne is varied group of diseases from teenage to cystic acne commonly
due to P. acnes which are quite susceptible to antimicrobial agents. Salicylic acid,
sulfur, benzoyl peroxide is widely preferred. Evaluation : Particle size of anti acne
compound greasiness Antimicrobial efficacy.

b) Skin tonic : used as Skin healing, promotion of tissue growth, Refreshing sensation.
Assay of active and label claimed ingredients.

c) Anti-ageing :
 Hair Care Products(Shampoo):
Foaming ability and foam stability: Cylinder shake method was used for determining
foaming ability. 50 ml of the 1% shampoo solution was put into a 250 ml graduated
cylinder and covered the cylinder with hand and shaken for 10 times.
The total volumes of the foam contents after 1 minute shaking were recorded.
The foam volume was calculated only. Immediately after shaking the volume of foam
at 1 minute intervals for 4 minutes were recorded.

Ross-miles foam column

2) Viscosity: this is also an important character and can be measured by viscometer

3) Effect on hair : this can be studied by half-head technique. In which half of the hair is
shampooed and the other half is used as control

4) Effect on skin and eyes: this can be measured by applying it on animals

5) pH : the pH of shampoo can be measured by pH meter and it should be between 6.0-

9.0

6) Stability studies: The thermal stability of formulations was studied byplacing in glass
tubes and they were placed in a humiditychamber at 45 C and 75% relative humidity.
Their appearanceand physical stability were inspected for a period of 3 monthsat
interval of one month.
Cleansing action : it can be tested on wool yarn in grease For this method place 5 gm
of wool yarn in grease in 200 ml of water containing 1 gm of shampoo in a flask,
shake the flask for 4 minute at rate of 50 times a minute. Remove the solution and
take out the sample. Dry it and weigh it. The loss in weight will indicate the amount of
grease removed which is the cleansing action of shampoo.

Fig: Hair after cleansing

Fig :Hair before cleansing
 Dental Care Product(Tooth paste & Tooth powders)
• Particle size : this can be determined by microscopic study of the particles or
by other means.
• Abrasiveness : the teeth are mechanically brushed with paste or powders using
tooth brush. The effect are studied by observation, mechanical (measurement with
micrometer gauge sensitive to 0.001 inch ) or other means (radioactive tracer techniques)
.• The pH of the aqueous solution : the pH of dispersion of 10% of the product in
water is determined by pH meter.
• Consistency : it is important that the product, paste, should maintain the
consistency to enable the product press out from the container study of viscosity is
essential for this. Rheology of powder is also important for proper flow of the powders
from the container.
• Volatile matters and moisture : a specific amount of product is taken in a
dish and drying is done till constant weight. Loss of weight will indicate amount of
moisture present in product.
 Personal Hygiene Products
1) Depilatories : It can be truly categorized as a cosmetic ,since it beautifies by
removing un slightly hair from certain parts of the body.
Evaluation
a) tensile kinetic method Stress decay caused by disulfide bond is measured using
commercial instrument such as
Tensile strength tester
An optical diameter
Electro balance
the time required to reduce the stress supported by hair by 95% was shown
to in vivo hair removal rate in commercial products.
b) HPLC method this method can distinguish between thioglcerol,thiolacticacid thiol
glycolic acid. the S-H group is coupled to 7 choloro-4nitro benzo-2oxo-1,diazole which
result in yellow derivative permitting HPLCdetection at 464 nm.
C) Thermo-mechanical method In this method thermo-mechanical analyzer is used to
measure the time at which ,a hair bundle underconstant stress & immersed in deplatory,
begin to stretch. the analyzer is programmed to observe the stretching &breaking of hair
bundle, attached to fiber tension probeaccessory. TLC &GLC METHODS are to identify
&approximately estimate the amount of active ingredientpresent.
 Personal Hygiene Products
2) Antiperspirant :
Antiperspirant it is therapeutic agent that actively reduce the amount of perspiration.

Evaluation
(a) Gravimetric method sweat collection is carried out in controlled temperature
rooms at 100 2 F and about 35%RH.. sweat collection are made during two successive
half an hour period using tared absorbent pads. A ratio sweat produced by right & left
axillae is determined in controlled condition. the effect of antiperspirant material on the
perspiration ratio of each individual is determined by comparing the post treatment ratio
with the subject average control ratio. reduction in
sweat rate (%) = post treatment ratio 100 average control ratio

(b) Hygrometry In this method cup is attached to the skin & water from the enclosed
area is evaporated by stream of dry gas. water content of this gas is monitored
& sweat rate is calculated.
 Personal Hygiene Products
3)Deodorant :It is cosmetic preparation that reduce the auxiliaryodour .
Evaluation In vivo & In vitro method.- Two principle method for in vivo evaluation of
deodorant efficiency are
1)determination of the effect of treatment in skin micro flora.
2) olfactory assessment of the effects on skin odour.
The different technique which are used to quantify micro flora
a)Tape stripping
b)Velvet replicate pads
c)Scrub technique
d) Pressurized spray method
Amongst method of evaluation deodorants method suggested by fredell & Longefellow
is widely usedOn the first day of test, odour of both axillae is recorded. A scale of 0 to 3
is used for recording the odour & direct sniffing is used for judging the odour. The product
is applied to the one axillae & nothing is applied to the control. After 6 hour both axillae
is again sniffed &the odour is recorded.The test may be repeated on the successive day
A pretest conditioning period is also recommended
for the success of test ,a definite characteristic odour Is imperative.
 Personal Hygiene Products
Shaving Preparations
Shaving preparations are product employed by the men to help inshaving.
Shaving preparation divided in to two group.
A) Preparation used before shaving.
B) Preparation used after shaving
Evaluation
1. spreadability
2. wetting
3. viscosity
4. foam texture
5. pH
6. compatibility & stability with perfume
7. effect on life of razor blade & reaction with the internal surface of collapsible tube
both metallic & plastic.
8. quality of the water
 Personal Hygiene Products
Shaving Preparations
Evaluation for both Lather & Brushless shaving cream To comply with Indian standard,
Specification for shaving cream as per drugs and cosmetic Act,1940 & rules
1945,shedule „S‟,

It is suggested that IS 9740:1981& its second amendment of October 1998 should

be referred.
1. consistency & texture
2. homogeneity
3. conformity of raw material
4. stability
5. effect on container
6. total fatty substance
7. water content
8. lathering power
9. free caustic alkali
 Personal Hygiene Products
Shaving Preparations
Evaluation of aerosol shaving cream
1. soap concentration
2. type of fatty acid
3. concentration of free fatty acid
4. type of polyols & its concentration
5. type of propellant & its concentration
6. pH 7. viscosity , density & stability of product.
After shave Lotion After shave lotion are non emulsified and mildly alcoholic solution.
Two type
a) alcoholic containg 5-70% alcohol
b) non alcoholic
Evaluation
1. Important test is alcohol content
2. stability of smell
3. cloud temperature
 Microbiological analysis of cosmetics
Cosmetics should be free from micro-organisms and consumer use.
The detection and elimination of microbial contamination of cosmetics is very important
to maximize shelf life.

• Ensure product quality, consistency and performance and to meet federal regulations.
Sample preparation for microbiological analysis

• For liquid: take 1 ml of liquid and diluted with 9 ml of modified letheen broth(MLB) in
screw-cap test tube.

• Solid and powders: weigh 1 gm of sample in to screw-cap test tube containing

1 ml sterile tween 80. disperse product in tween 80 with sterile spatula. Add 8 ml sterile
MLB and mix thoroughly.

• Wax / fatty products (lipsticks): weigh 10 gm of sample in to sterile tween 20. disperse
with a sterile spatula to form a paste. Add 78 ml sterile MLB and mix thoroughly.
Method for microbiological analysis
• Pour-plate technique: The sample
should be diluted successively with
sterile water. The agar medium is
maintained in molten state at 45˚c.
1 ml of diluted sample is added to
sterile petri dish to which is then
poured 9 ml of sterile, cool agar
medium. The contents are
thoroughly mixed and allowed to
solidify. The dish is incubated at
suitable temperature and
conditions. After few days, different
kinds of microbe grow as separate
colonies.
 Method for microbiological analysis
• Spread plate technique: An
aliquot of the diluted sample is
placed an to the agar surface and is
spread uniformly with a sterile bent
rod. Incubate it at suitable
temperature and condition. After
few days, different kinds of
microbes grow as separate
colonies.
 Method for microbiological analysis
• Membrane filtration Method :
A known amount of pretreated material or its
dilution is passed trough membrane filter
assembly. Wash it 3 successive times each
of 100 ml ofbuffered Nacl-peptone solution.
Transfer the membrane on the surface of
solidagar medium in a sterile Petri dish. The
dish is incubated at suitable temperature and
conditions. After few days different kinds of
microbe grow as separate colonies. The
Indian standards (is:11377-1985)prescribes
that bacterial count of a cosmetics should not
exceed 1000 microorganism per 1 gm of
cosmetic and there should not any
pathogens.
Chemical analysis of cosmetics

Cosmetic analysis. Features and most suitable properties of the analytical methods.
 Chemical analysis of cosmetics
Chemical analysis of cosmetics is very important to ensure that only permitted ingredient
are added to the product, information on the label is correct or not, and to help in
forensic investigation.
General methods

i. Determination of methanol in relation to ethanol or 2-propanol by gas chromatography.

ii. Determination of dichloromethane and 1,1,1 trichloroethane by gas chromatography.

iii. Determination of chlorobutanol by gas chromatography.

iv. Determination of hexachlorophene by gas chromatography.

v. Determination of water by gas chromatography.

vi. Determination of propylene glycol by gas chromatography. thoroughly.

 Deodorants and antiperspirants

i. Aluminium and zinc in deodorants by gravimetric method or by flame atomic

absorption spectroscopy.

ii. Zirconium in anti perspirants by colorimetric method or by flame atomic absorption

spectrometry.

iii. Boric acid in deodorants and anti perspirants by ion-exchange method.

iv. Chlorides and sulfates in deodorants by gravimetric method.

v. Methenamine and urea in deodorants by titrimetric method.

 Hair preparation

i.Quinine in shampoo and in hair lotion by HPLC

ii. oxalic acid and alkaline salt in hair- care product by filtration.

iii. Free sodium and potassium hydroxide in hair straightener by filtration.

iv. Mercapto acetic acid in hair-waving and in hair-straightening by iodometric titration

or gas chromatography.

v. Selenium disulphide as selenium in anti-dandruff shampoos by atomic absorption

spectrometry.
 Analytical Methods

Hair Dyes

• According to how long-lasting they are three types

a. temporary,
b. semi-permanent,
c. permanent hair colours

Methods use for quantitative and semi quantitative determination of hair dyes are

i. one- or two-dimensional thin-layer chromatography,

ii. gravimetrically,
iii. colorimetrically
iv. Thin-layer chromatography (TLC),
v. gas chromatography (GC),
vi. liquid chromatography (LC)
 Analytical Methods
Tooth pastes
i. Chloroform and chlorates of
alkali metals in tooth paste by Sun screen lotions
gas chromatography.
ii. Total fluorine in dental
creams by gas
chromatography.
Creams and pastes
i. Nitrite creams and pastes
by colorimetric method.
Vanishing creams
i. Water by karl-fiesher
titrationii. Ashiii. Chloroform
soluble material by GC

Photo stability Testing

 Analytical Methods of Colouring Agents
in Decorative and other Cosmetics

• Decorative cosmetics are

principally used to beautify.

• The different types of

decorative cosmetics include
foundations, lipsticks,
glosses, mascaras, nail
lacquers and powders.

Fig:General approach to the preparative

separation of dyes by high-speed
countercurrent chromatography
 Analytical Methods
Determination of colouring agents in cosmetic products
• Thin-layer chromatography
• Liquid chromatography
• Spectrophotometry
• Other methods dyes in lipstick using micellar electrokinetic capillary
chromatography (MEKC) with diode array UV detection
Analytical Methods Preservatives in Cosmetics
• preservatives belonging to different chemical classes therefore, multicomponent
analysis methods are required ,like
1. ion-pair and reversed-phase LC with UV/Vis detection,
2. Thin layer chromatography (TLC)
3. capillary electrophoresis (CE)
4. capillary zone electrophoresis (CZE)
5. gas chromatography (GC) with flame ionization detector (FID),
electron capture detector (ECD) or mass spectrometry (MS) detector
used for preservative determination
 Analysis of perfumes
• ultra violet/visible spectrometry (UV/VIS),
• infrared spectrometry (IR)
• nuclear magnetic resonance (NMR),
• gas chromatography (GC), both by injection or
in headspace (HS) mode
• liquid chromatography (LC) and thin-layer
chromatography (TLC) have also been applied
for quantitative and/or qualitative purposes in
perfume analysis,
• GC-MS
• LC-UV/VIS
• GC-FID Fig: Headspace micro- 57
extraction
• LC-FL (TLC with fluorimetric)
• TLC with fluorescence densitometry (FD)
 References:
1. Text book of “Cosmetics Formulation, manufacturing & Quality control”, by P. P.
Sharma 4th edition, Vandana Publications Pvt. ltd.
2. “Cosmetic Technology”, by Sanju Nanda, Arun
Nanda,RoopK.Khar1stEdition,BirlaPublications
3. “Cosmetics”, by Sagarin,Volume 1 & 3

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Thank You