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SEMINAR ON
FORMULATION & EVALUATION
OF SHAMPOO
Presented By
Manas Boxi
M.Pharm (Pharmaceutics)
1st Semester
Regd. No. A-05
Roland Institute of Pharmaceutical Sciences
CONTENTS
1. Introduction
2. Types of Shampoos
3. Product Ingredients
4. Formulation
5. Evaluation of Shampoos
6. References
Introduction
• Definition: A shampoo is a preparation of a surfactant (i.e.
surface active material) in a suitable form – liquid, solid or
powder – which when used under the specified conditions
will remove surface grease, dirt, and skin debris from the hair
shaft and scalp without adversely affecting the user.
• Requirements of a Shampoo:
1. It should effectively and completely remove dust or soil,
excessive sebum or other fatty substances and loose corneal
cells from the hair.
2. It should produce a good amount of foam to satisfy the
psychological requirements of the user.
3. It should be easily removed on rinsing with water.
4. It should leave the hair non-dry, soft, lustrous with good
manageability and minimum fly away.
5. It should impart a pleasant fragnance to the hair.
6. It should not cause any side-effects / irritation to skin or eye.
7. It should not make the hand rough and chapped.
Types of Shampoo
Shampoos are of the following types:
 Powder Shampoo
 Liquid Shampoo
 Lotion Shampoo
 Cream Shampoo
 Jelly Shampoo
 Aerosol Shampoo
 Specialized Shampoo
• Conditioning Shampoo
• Anti- dandruff Shampoo
• Baby Shampoo
• Two Layer Shampoo
PRODUCT INGREDIENTS
• Surfactants are the main component of shampoo. Mainly anionic
surfactants are used.
• The raw materials used in the manufacture of shampoos are:
1. Principal surfactants: Provide detergency and foam.
2. Secondary surfactants: Improve detergency, foam and hair
condition.
3. Other additives.
CLEANSING ACTION OF SHAMPOO
A surfactant consists of two part- one hydrophilic (water loving)
while the other is hydrophobic in nature.
Surfactants
 Anionic surfactants are mostly used (good foaming
properties). The hydrophilic portion carries a negative
charge which results in superior foaming, cleaning
and end result attributes.
 Non-ionic surfactants have good cleansing properties
but do not have sufficient foaming power.
 Cationic surfactants are toxic and are hence not used.
However, they may be used in low concentration in
hair conditioners.
 Ampholytics, being expensive, are generally not used.
However, they are mainly used as secondary
surfactants and good hairconditioners.
ANIONIC SURFACTANTS
CLASS EXAMPLE COMMENT
Alkyl benzene
sulfonates
Sodium dodecyl benzene
sulfonate
Tend to yield an “airy” or low density foam
and often are drying to the hair
Primary alkyl sulfates Lauric acid, stearic acid and
their salts
Good lathering effect in hard water, free from
rancidity, easy to wash.
Secondary alcohol
sulfates
Sodium sec-lauryl sulfate Low cost, dispersing and emulsifying action,
dissapointing as detergets and shampoo
components
Alkyl benzene
polyoxyethylene
sulfonates
Triton X200 Stable in acid or alkaline solution, excellent
emulsifier, detergent and wetting agent;
extremely stable at pH of skin
Sulfated
monoglycerides
Lauric monoglyceride
ammonium sulfate
Stable in hard water
Alkyl ether sulfates Derivatives of lauryl alcohol
ether with PEG
Good cleansers, act as solvents for non polar
additives
Sarcosines Lauroyl and cocoyl
sarcosines
Excellent foaming and conditioning action
Sulfosuccinates Aerosol OT Less irritating to skin and eye (baby shampoo)
Maypon Protalbinic and lysalbinic
acid derivatives (maypon 4C)
Hydrolysation product of proteins with fatty
acid chlorides in presence of alkali
NON-IONIC SURFACTANTS
CLASS EXAMPLE COMMENTS
Fatty acid
alkanolamides (should
not be used > 15%)
Lauric monoethanolamide Improves solubility of SLS
Stearic ethanolamide Pearlescent thickener
Oleic ethanolamides Hair conditioning agents
Polyalkoxylated
derivatives
Ethoxylated fatty alcohols Stable in wide range of pH; stabilizing
emulsifying and opacifying properties
Block polymers (pluronics) Good rinsability, can be used in high %
Sorbitol esters (TWEENS) Solubilizers and emulsifiers, used in baby
shampoos
Amine oxides Coconut and dodecyl
dimethyl amine oxides
Foam booster and anti-static agents
AMPHOTERIC SURFACTANTS
N-alkyl aminoacids β – aminoacid derivatives Foaming agents
Aspargine derivatives Compatible with both anionic and
cationic surfactants
Betains Amido betains High foaming properties, mild.
Alkyl imidazoline MIRANOLTM Baby shampoos
ADDITIVES
 Conditioning agents: Lanolin, mineral oil, herbal extracts, egg
derivatives.
 Foam builders: Lauroyl monoethanolamide, sarcosinates
 Viscosity modifiers :
 Electrolytes – NH4Cl, NaCl
 Natural gums – Gum Karaya, tragacanth, alginates
 Cellulose derivatives – Hydroxy ethyl cellulose, methyl cellulose
 Carboxy vinyl polymers – Carbopol 934
 Others – PVP, phosphate esters.
 Sequestering agents : EDTA
 Opacifying agents : Alkanolamides of higher fatty acids, propylene
glycol, Mg, Ca and Zn salts of stearic acid, spermaceti, etc.
 Clarifying agents :
 Solubilizing alcohols – ethanol, isopropanol
 Phosphates –
 Non-ionic solubilizers – polyethoxyated alcohols and esters.
ADDITIVES
 Perfumes : Herbal, fruity or floral fragnances.
 Preservatives : Methyl and propyl paraben,
formaldehyde (most effective).
 Anti-dandruff agents: The shampoos contain small
amount of these actives, which are in contact with
the scalp for only a short time. In order to be
effective the active ingredient must work in the oil-
water environment of the scalp and must be readily
substantive to the scalp for continuing activity.
Ex: Selenium sulfide, zinc pyrithone, salicylic acid.
FORMULATIONS
POWDER SHAMPOO
Henna powder 5%
Soap powder 50%
Sodium carbonate 22.5%
Potassium carbonate 7.5%
Borax 15%
Perfume q.S
LIQUID SHAMPOO
SLS 40%
NaCl (to desired viscosity) 2-4%
Water Upto 100%
Perfume, color, preservatives q.s
LOTION SHAMPOO
TLS 35%
Glyceryl monostearate 2%
Magnesium stearate 1%
Water Upto 100%
Color q.s
Perfume, preservatives q.s
FORMULATIONS
CREAM SHAMPOO
SLS 38%
Cetyl alcohol 7%
Water Upto 100%
Color, perfume q.S
Preservative q.s
JELLY SHAMPOOS
Alkyl dimethyl benzalkonium chloride 15%
TLS (40%) 28%
Coconut ditethanolamide 7%
HPMC 1%
Water Upto 100%
Color, perfume, preservative q.s
AEROSOL SHAMPOO
TLS 60%
Coconut diethanolamide 2%
Water Upto 90%
Propellent 10%
Color, perfume, preservative q,.s
FORMULATIONS
CONDITIONING SHAMPOOS
Steryl dimethyl benzyl
ammonium chloride
5.5%
Ethylene glycol
monostearate
2%
Cetyl alcohol 2.5%
Water Upto 100%
Color, perfume, preservative q.s
TWO LAYER SHAMPOO
SLS 27%
Cocamidopropylamine oxide 5%
Lauramine DEA 1%
Lactic acid (50%) 1%
Formaldehyde 0.1%
BABY SHAMPOO
Magnesium lauryl sulfate (27.5%) 11%
Cocamidopropyl betaine (30%) 5%
Polysorbate 20 1%
PEG 600 3.5%
Perfume q.S
Preservative q.S
Citric acid To pH 6
Color q.S
Water (deionised); Aqua (INCI) To 100%
FORMULATIONS
ANTI-DANDRUFF SHAMPOO
Thymol 0.05%
Menthol 0.1%
Camphor 0.1%
TLS 55%
Water upto 100
Color, perfume, preservative q.s
ANTI-DANDRUFF SHAMPOO
Selenium sulfide 2.5%
Bentonite 5%
SLS paste 35%
Water upto 100
Color, perfume, preservative q.s
FORMULATION
HERBAL SHAMPOO
Natural essential oil blend 0.5%
Cyamopsis tetragonoloba (Guar Gum) 1%
Camellia sinensis (Green Tea) extract 2%
Glycerin 1%
Hydrolysed wheat protein 2.5%
Salvia officinalis (Sage) leaf extract 1.5%
Salvia officinalis (Sage) 1.5%
Glyceryl oleate 1%
Polysorbate 20 0.5%
Potassium sorbate 5%
Aloe barbadensis (Aloe vera) extract 0.5%
Arctium minus (Burdock) root extract 0.5%
Disodium coco-glucoside sulfosuccinate 0.5%
Preservatives q.s.
Water Upto 100%
Evaluation of Shampoos
 Performance characteristics
 Foam and foam stability
 Detergency and cleaning action
Effect of water hardness
Surface Tension and wetting
Surfactant content and analysis
 Rinsing
 Conditioning action
 Softness
 Luster
 Lubricity
 Body, texture and set retention
 Irritation and toxicity
 Dandruff control
 Microbiological assay
 Eye irritancy test
Product characteristics
Fragnance
Colour
Consistency
Package
1. Foam and foam stability:
• The Ross-Miles foam column test is accepted. 200 ml of surfactant
solution is dropped into a glass column containing 50ml of the same
solution. The height of the foam generated is measured immediately
and again after a specified time interval, and is considered
proportional to the volume.
• Barnett and Powers developed a latherometer to measure the effect
of variables such as water hardness, type of soil and quantity of soil on
foam speed, volume and stability.
• Fredell and Read titrated actual standard oiled heads of hair with
additive increments of shampoo until a persistent lather end point
appeared.
2. Detergency and cleaning action:
• Cleansing power is evaluated by the method of Barnet and Powers
• 5gm sample of soiled human hair is placed at 35°c in 200 cc of water
containing of 1 gm of shampoo.
• The flask is shaken 50 times a minute for 4 minutes. Then washed
once again with sufficient amount of water, then after filter the hair
dried and weighed.
• The amount of soil is removed under these condition is calculated.
3. Wetting Action:
• Canvas disk sinking test: A mount veron cotton duck # 6 canvas disk 1
inch in diameter, is floated on the surface of a solution, and the time
required for it to sink is measured accurately.
4. Rinsing:
• Skilled beauticians are employed to make comparisons on the
performance of several shampoos.
5. Conditioning Action:
• Conditioning action is a difficult property to assess. This is because it is
basically dependent on subjective appraisal.
• No method has been published for measuring conditioning action.
• The degree of conditioning given to hair is ultimately judged by
shampoo user who is making the evaluation on the basis of past
experience and present expectations.
6. Microbiological assay:
 PREPARATION OF PRE-INOCULUM Take the loopful culture of
staphylococcus aureus (ATCC6532) aseptically and transfer to
sterilized and cooled 100 ml SCDM (broth).
• Mix well. Incubate the broth at 37oC for 24 hrs.
 PREPARATION OF MEDIA Soya bean casein digests medium, soya
bean casein digest agar and nutrient agar.
 PREPARATION OF POUR PLATES Sterilized SCD agar (100 ml) is
cooled to 40°C and mixed with 5 ml of 24 hrs old pre inoculated
culture.
• This is immediately poured in plates (340 ml each) and allows to
set.
 MAKING THE WELLS ON AGAR PLATES The wells are dig on agar
plates with sterilised well digger aseptically.
• Take 100µml of each sample, add to well aseptically. Incubate the
plates at 37oC for 24 hrs to 48 hrs.
• Observe the effectiveness of sample on culture growing on the agar
plate and we can see the effectiveness of sample in the form of
zone of inhibition around each well containing different sample.
7. Evaluation of eye irritancy:
• The test calls for dropping 0.1 ml of liquid shampoo in the
conjunctiva sac of one eye of the rabbit , the other eye serving as
control.
• In the case of the first three animals, the treated eye remains
unwashed. Since washing the eye may or may not alleviate
symptoms of injury.
• The six remaining animals are divided into two equal groups.
• In the first of these groups eyes instilled with the substances are
washed with 20 ml of lukewarm water two seconds after
treatment and in the second group after instillation.
• Readings are then made at 24, 48 and 72 hr and again four and
seven days after treatment.
• If the lesions have not cleared up in seven days the test material is
considered as severe irritant.
8. Viscosity:
• Viscosity of the liquid shampoo is determined using a Brookefield
viscometer
• 100 mL of the shampoo is taken in a beaker and the spindle is
dipped in it for about 5 min and then the reading is taken.
References
1. Balsam, S.M., Gershon, S.D., Rieger, M.M., Sagarin, E., and Strianse, S.J.: COSMETICS–
Science and Technology, 2nd edition, Vol-2, John Wiley India, New Delhi, 2008
2. Barel, A.O., Paye, M., and Maibach, H.I.: Handbook of Cosmetic Science and
Technology, 3rd Edition, Informa Healthcare, New York.
3. Sharma, P.P.: COSMETICS - Formulation, Manufacturing and Quality Control, 4th
Edition, Vandana Publishers Pvt. Ltd., New Delhi, March 1998.
4. Butler, H.: POUCHER’S – Perfumes, Cosmetics & Soaps, 10th Edition, Springer,
Cockermouth, Cumbria, USA, 2000.
5. Salador, A., and Chisvert, A.: Analysis of cosmetic products, Elsevier, New York, 2006.
6. Ross, J., and Miles, G.D.: An application for comparison of foaming properties of
soaps and detergents, Oil and Soap, 1941.
7. Mittal,: A Handbook of Cosmetics
8. Fredell, W.G., and Powers, D.H.: Factors attributing to the performance of shampoos
and to consumer acceptance, Proc. Sci. Sec., 1955.
9. Rajkumar, K. J., Invitro evaluation of shampoos.
10. www.cosmeticdatabase.com
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Evalulation

  • 1. SEMINAR ON FORMULATION & EVALUATION OF SHAMPOO Presented By Manas Boxi M.Pharm (Pharmaceutics) 1st Semester Regd. No. A-05 Roland Institute of Pharmaceutical Sciences
  • 2. CONTENTS 1. Introduction 2. Types of Shampoos 3. Product Ingredients 4. Formulation 5. Evaluation of Shampoos 6. References
  • 3. Introduction • Definition: A shampoo is a preparation of a surfactant (i.e. surface active material) in a suitable form – liquid, solid or powder – which when used under the specified conditions will remove surface grease, dirt, and skin debris from the hair shaft and scalp without adversely affecting the user. • Requirements of a Shampoo: 1. It should effectively and completely remove dust or soil, excessive sebum or other fatty substances and loose corneal cells from the hair. 2. It should produce a good amount of foam to satisfy the psychological requirements of the user. 3. It should be easily removed on rinsing with water. 4. It should leave the hair non-dry, soft, lustrous with good manageability and minimum fly away. 5. It should impart a pleasant fragnance to the hair. 6. It should not cause any side-effects / irritation to skin or eye. 7. It should not make the hand rough and chapped.
  • 4. Types of Shampoo Shampoos are of the following types:  Powder Shampoo  Liquid Shampoo  Lotion Shampoo  Cream Shampoo  Jelly Shampoo  Aerosol Shampoo  Specialized Shampoo • Conditioning Shampoo • Anti- dandruff Shampoo • Baby Shampoo • Two Layer Shampoo
  • 5. PRODUCT INGREDIENTS • Surfactants are the main component of shampoo. Mainly anionic surfactants are used. • The raw materials used in the manufacture of shampoos are: 1. Principal surfactants: Provide detergency and foam. 2. Secondary surfactants: Improve detergency, foam and hair condition. 3. Other additives. CLEANSING ACTION OF SHAMPOO A surfactant consists of two part- one hydrophilic (water loving) while the other is hydrophobic in nature.
  • 6. Surfactants  Anionic surfactants are mostly used (good foaming properties). The hydrophilic portion carries a negative charge which results in superior foaming, cleaning and end result attributes.  Non-ionic surfactants have good cleansing properties but do not have sufficient foaming power.  Cationic surfactants are toxic and are hence not used. However, they may be used in low concentration in hair conditioners.  Ampholytics, being expensive, are generally not used. However, they are mainly used as secondary surfactants and good hairconditioners.
  • 7. ANIONIC SURFACTANTS CLASS EXAMPLE COMMENT Alkyl benzene sulfonates Sodium dodecyl benzene sulfonate Tend to yield an “airy” or low density foam and often are drying to the hair Primary alkyl sulfates Lauric acid, stearic acid and their salts Good lathering effect in hard water, free from rancidity, easy to wash. Secondary alcohol sulfates Sodium sec-lauryl sulfate Low cost, dispersing and emulsifying action, dissapointing as detergets and shampoo components Alkyl benzene polyoxyethylene sulfonates Triton X200 Stable in acid or alkaline solution, excellent emulsifier, detergent and wetting agent; extremely stable at pH of skin Sulfated monoglycerides Lauric monoglyceride ammonium sulfate Stable in hard water Alkyl ether sulfates Derivatives of lauryl alcohol ether with PEG Good cleansers, act as solvents for non polar additives Sarcosines Lauroyl and cocoyl sarcosines Excellent foaming and conditioning action Sulfosuccinates Aerosol OT Less irritating to skin and eye (baby shampoo) Maypon Protalbinic and lysalbinic acid derivatives (maypon 4C) Hydrolysation product of proteins with fatty acid chlorides in presence of alkali
  • 8. NON-IONIC SURFACTANTS CLASS EXAMPLE COMMENTS Fatty acid alkanolamides (should not be used > 15%) Lauric monoethanolamide Improves solubility of SLS Stearic ethanolamide Pearlescent thickener Oleic ethanolamides Hair conditioning agents Polyalkoxylated derivatives Ethoxylated fatty alcohols Stable in wide range of pH; stabilizing emulsifying and opacifying properties Block polymers (pluronics) Good rinsability, can be used in high % Sorbitol esters (TWEENS) Solubilizers and emulsifiers, used in baby shampoos Amine oxides Coconut and dodecyl dimethyl amine oxides Foam booster and anti-static agents AMPHOTERIC SURFACTANTS N-alkyl aminoacids β – aminoacid derivatives Foaming agents Aspargine derivatives Compatible with both anionic and cationic surfactants Betains Amido betains High foaming properties, mild. Alkyl imidazoline MIRANOLTM Baby shampoos
  • 9. ADDITIVES  Conditioning agents: Lanolin, mineral oil, herbal extracts, egg derivatives.  Foam builders: Lauroyl monoethanolamide, sarcosinates  Viscosity modifiers :  Electrolytes – NH4Cl, NaCl  Natural gums – Gum Karaya, tragacanth, alginates  Cellulose derivatives – Hydroxy ethyl cellulose, methyl cellulose  Carboxy vinyl polymers – Carbopol 934  Others – PVP, phosphate esters.  Sequestering agents : EDTA  Opacifying agents : Alkanolamides of higher fatty acids, propylene glycol, Mg, Ca and Zn salts of stearic acid, spermaceti, etc.  Clarifying agents :  Solubilizing alcohols – ethanol, isopropanol  Phosphates –  Non-ionic solubilizers – polyethoxyated alcohols and esters.
  • 10. ADDITIVES  Perfumes : Herbal, fruity or floral fragnances.  Preservatives : Methyl and propyl paraben, formaldehyde (most effective).  Anti-dandruff agents: The shampoos contain small amount of these actives, which are in contact with the scalp for only a short time. In order to be effective the active ingredient must work in the oil- water environment of the scalp and must be readily substantive to the scalp for continuing activity. Ex: Selenium sulfide, zinc pyrithone, salicylic acid.
  • 11. FORMULATIONS POWDER SHAMPOO Henna powder 5% Soap powder 50% Sodium carbonate 22.5% Potassium carbonate 7.5% Borax 15% Perfume q.S LIQUID SHAMPOO SLS 40% NaCl (to desired viscosity) 2-4% Water Upto 100% Perfume, color, preservatives q.s LOTION SHAMPOO TLS 35% Glyceryl monostearate 2% Magnesium stearate 1% Water Upto 100% Color q.s Perfume, preservatives q.s
  • 12. FORMULATIONS CREAM SHAMPOO SLS 38% Cetyl alcohol 7% Water Upto 100% Color, perfume q.S Preservative q.s JELLY SHAMPOOS Alkyl dimethyl benzalkonium chloride 15% TLS (40%) 28% Coconut ditethanolamide 7% HPMC 1% Water Upto 100% Color, perfume, preservative q.s AEROSOL SHAMPOO TLS 60% Coconut diethanolamide 2% Water Upto 90% Propellent 10% Color, perfume, preservative q,.s
  • 13. FORMULATIONS CONDITIONING SHAMPOOS Steryl dimethyl benzyl ammonium chloride 5.5% Ethylene glycol monostearate 2% Cetyl alcohol 2.5% Water Upto 100% Color, perfume, preservative q.s TWO LAYER SHAMPOO SLS 27% Cocamidopropylamine oxide 5% Lauramine DEA 1% Lactic acid (50%) 1% Formaldehyde 0.1% BABY SHAMPOO Magnesium lauryl sulfate (27.5%) 11% Cocamidopropyl betaine (30%) 5% Polysorbate 20 1% PEG 600 3.5% Perfume q.S Preservative q.S Citric acid To pH 6 Color q.S Water (deionised); Aqua (INCI) To 100%
  • 14. FORMULATIONS ANTI-DANDRUFF SHAMPOO Thymol 0.05% Menthol 0.1% Camphor 0.1% TLS 55% Water upto 100 Color, perfume, preservative q.s ANTI-DANDRUFF SHAMPOO Selenium sulfide 2.5% Bentonite 5% SLS paste 35% Water upto 100 Color, perfume, preservative q.s
  • 15. FORMULATION HERBAL SHAMPOO Natural essential oil blend 0.5% Cyamopsis tetragonoloba (Guar Gum) 1% Camellia sinensis (Green Tea) extract 2% Glycerin 1% Hydrolysed wheat protein 2.5% Salvia officinalis (Sage) leaf extract 1.5% Salvia officinalis (Sage) 1.5% Glyceryl oleate 1% Polysorbate 20 0.5% Potassium sorbate 5% Aloe barbadensis (Aloe vera) extract 0.5% Arctium minus (Burdock) root extract 0.5% Disodium coco-glucoside sulfosuccinate 0.5% Preservatives q.s. Water Upto 100%
  • 16. Evaluation of Shampoos  Performance characteristics  Foam and foam stability  Detergency and cleaning action Effect of water hardness Surface Tension and wetting Surfactant content and analysis  Rinsing  Conditioning action  Softness  Luster  Lubricity  Body, texture and set retention  Irritation and toxicity  Dandruff control  Microbiological assay  Eye irritancy test Product characteristics Fragnance Colour Consistency Package
  • 17. 1. Foam and foam stability: • The Ross-Miles foam column test is accepted. 200 ml of surfactant solution is dropped into a glass column containing 50ml of the same solution. The height of the foam generated is measured immediately and again after a specified time interval, and is considered proportional to the volume. • Barnett and Powers developed a latherometer to measure the effect of variables such as water hardness, type of soil and quantity of soil on foam speed, volume and stability. • Fredell and Read titrated actual standard oiled heads of hair with additive increments of shampoo until a persistent lather end point appeared. 2. Detergency and cleaning action: • Cleansing power is evaluated by the method of Barnet and Powers • 5gm sample of soiled human hair is placed at 35°c in 200 cc of water containing of 1 gm of shampoo. • The flask is shaken 50 times a minute for 4 minutes. Then washed once again with sufficient amount of water, then after filter the hair dried and weighed. • The amount of soil is removed under these condition is calculated.
  • 18. 3. Wetting Action: • Canvas disk sinking test: A mount veron cotton duck # 6 canvas disk 1 inch in diameter, is floated on the surface of a solution, and the time required for it to sink is measured accurately. 4. Rinsing: • Skilled beauticians are employed to make comparisons on the performance of several shampoos. 5. Conditioning Action: • Conditioning action is a difficult property to assess. This is because it is basically dependent on subjective appraisal. • No method has been published for measuring conditioning action. • The degree of conditioning given to hair is ultimately judged by shampoo user who is making the evaluation on the basis of past experience and present expectations.
  • 19. 6. Microbiological assay:  PREPARATION OF PRE-INOCULUM Take the loopful culture of staphylococcus aureus (ATCC6532) aseptically and transfer to sterilized and cooled 100 ml SCDM (broth). • Mix well. Incubate the broth at 37oC for 24 hrs.  PREPARATION OF MEDIA Soya bean casein digests medium, soya bean casein digest agar and nutrient agar.  PREPARATION OF POUR PLATES Sterilized SCD agar (100 ml) is cooled to 40°C and mixed with 5 ml of 24 hrs old pre inoculated culture. • This is immediately poured in plates (340 ml each) and allows to set.  MAKING THE WELLS ON AGAR PLATES The wells are dig on agar plates with sterilised well digger aseptically. • Take 100µml of each sample, add to well aseptically. Incubate the plates at 37oC for 24 hrs to 48 hrs. • Observe the effectiveness of sample on culture growing on the agar plate and we can see the effectiveness of sample in the form of zone of inhibition around each well containing different sample.
  • 20. 7. Evaluation of eye irritancy: • The test calls for dropping 0.1 ml of liquid shampoo in the conjunctiva sac of one eye of the rabbit , the other eye serving as control. • In the case of the first three animals, the treated eye remains unwashed. Since washing the eye may or may not alleviate symptoms of injury. • The six remaining animals are divided into two equal groups. • In the first of these groups eyes instilled with the substances are washed with 20 ml of lukewarm water two seconds after treatment and in the second group after instillation. • Readings are then made at 24, 48 and 72 hr and again four and seven days after treatment. • If the lesions have not cleared up in seven days the test material is considered as severe irritant. 8. Viscosity: • Viscosity of the liquid shampoo is determined using a Brookefield viscometer • 100 mL of the shampoo is taken in a beaker and the spindle is dipped in it for about 5 min and then the reading is taken.
  • 21. References 1. Balsam, S.M., Gershon, S.D., Rieger, M.M., Sagarin, E., and Strianse, S.J.: COSMETICS– Science and Technology, 2nd edition, Vol-2, John Wiley India, New Delhi, 2008 2. Barel, A.O., Paye, M., and Maibach, H.I.: Handbook of Cosmetic Science and Technology, 3rd Edition, Informa Healthcare, New York. 3. Sharma, P.P.: COSMETICS - Formulation, Manufacturing and Quality Control, 4th Edition, Vandana Publishers Pvt. Ltd., New Delhi, March 1998. 4. Butler, H.: POUCHER’S – Perfumes, Cosmetics & Soaps, 10th Edition, Springer, Cockermouth, Cumbria, USA, 2000. 5. Salador, A., and Chisvert, A.: Analysis of cosmetic products, Elsevier, New York, 2006. 6. Ross, J., and Miles, G.D.: An application for comparison of foaming properties of soaps and detergents, Oil and Soap, 1941. 7. Mittal,: A Handbook of Cosmetics 8. Fredell, W.G., and Powers, D.H.: Factors attributing to the performance of shampoos and to consumer acceptance, Proc. Sci. Sec., 1955. 9. Rajkumar, K. J., Invitro evaluation of shampoos. 10. www.cosmeticdatabase.com