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Journal of Biotechnology and Biosafety !
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Research art i c l e
REVIEW ON HAIR, DANDRUFF, SHAMPOO AND EXPERIMENTAL,
ANTIMICROBIAL ACTIVITY OF PIROCTONE OLAMINE WHICH IS USED AS
ACTIVE IN FORMULATION ANTI DANDRUFF SHAMPOO
______________________________________
Nitesh Rajput1 Vishal Chinchamalatpure
2*
______________________________________
1Senoir Manager,R & D, Vitro Med Health Care,
Jaipur.
2Executive, R & D Cosmetic, Sri Sri Ayurveda
Trust, Bangalore-560082.
*Corresponding author email id:
vishalchinchamalatpure@gmail.com
ABSTRACT: In recent days, so many people are facing
problems of dandruff. Dandruff causes lots of problems such as
itching, scaling, irritation etc. Due to dandruff hair fall takes
place giving dull and diminish look to hair. It is reported that
Dandruff and hairfall is because of anti-fungal, anti-bacterial
property. Hence the present work is a humble attempt to
evaluate the definition of hair dandruff Melassezia furfur,
Pityrosporum ovale, and candida albicans, which are
responsible for the formation of dandruff and shampoo.
The quality control of piroctone olamine is of paramount
importance for justifying their quality and integrity for
acceptance in modern system of cosmeceuticals. Hence the
procured sample of the chemical material was first evaluated
for their quality by determining – colour, odour, pH, solubility,
melting point, residue on ignition and % purity. The values
obtained within the standard values and hence it was concluded
that the procured material was pure to be used for the further
experimentation.
Key words: hair, dandruff, shampoo, Melassezia furfur, Pityrosporum ovale,, candida albicans, piroctone olamine
________________________________________________________________________________________________________
REVIEW ON HAIR, DANDRUFF, SHAMPOO
INTRODUCTION:
Hair
Hairs, or pili, are present on most skin surfaces except the
palm, palmer surfaces of the feet. In adults, hair usually is
most heavily distributed across the scalp, in the eyebrows,
in the axillae (armpits), and around the external genitalia.
Genetic and hormonal influences largely determine the
thickness and the pattern of the hair distribution. (Tortora
G. J., Derrickson B. H, 2009, p-155)
The functions of hair (Phate R. P, 1994, p-244)
• Protection of scalp from the external damage.
• To give attractive look and shape to the face and to
enhance the facial expression.
• Healthy hairs enhance beauty.
• Protect the skin against mechanical trauma and
heat loss or high temperature.
• Protect the scalp against UV radiation.
Anatomy of hair (Tortora G. J., Derrickson B. H, 2009)
Each hair is composed of columns of dead, keratinized
epidermal cells bonded together by extracellular proteins.
The shaft is the superficial portion of the hair, which
projects above the surface of the skin. The root is the
portion of the hair deep to the shaft that penetrates into the
dermis, and sometimes into the subcutaneous layer of the
skin.
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A hair is a specialized outgrowth of part of the skin called
the epidermis. Outermost lines from the inner root sheath
and inner cell from the further hair shaft i.e. the part of hair
emerging from the skin is called hair shaft and part which is
in the skin is hair root. The epidermis invigilated into the
dermis to form a tubal as pocket called hair follicle. The
sebaceous gland open directly into the upper part of the
follicle, this gland secretes sebum which lubricate and
protects the scalp and hair.
A smooth muscle called arrector pili muscle which is under
autonomic control is connected near the center of hair
follicle and runs at an angle towards the epidermis.
The center of hair bulb is invigilated to form the dermal
papilla for generation and for growth of hair. The cells of
hair in contact with dermal papilla are called hair matrix and
are responsible for actual production of hair
Hair has two distinct parts, the hair follicle and the hair
shaft.
Hair follicle (Phate R. P, 1994,p-246)
Each hair on human body grows form a hair follicle, a tiny
saclike hole in human skin. At the bottom of the each
follicle is a cluster of special cells that produce to make new
hair cells.
The new cells that are produced are added on at the root of
the hair causing the hair to grow longer. The active hair
follicle is divided into 3-segment. Upper segment consist of
infundibulum, middle isthmus lies between the duct of the
sebaceous gland and the attachment of arrector pili muscles.
Lower inferior segment is hair bulb. The center of hair bulb
is invigilated at the lower end to form the dermal papilla.
These cells in contact with dermal papilla are called hair
matrix. These are responsible for actual production of hair.
It contains melanocytes responsible for color of hair.
Fig. No. 1. Hair Shaft ( http://tinyurl.com/cfbhs22)
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Hair shaft (Tortora G. J., Derrickson B. H, 2009, p-157)
Figure no. 1 shows a longitudinal and transverse section
through a hair shaft. The shaft and the root of the hair both
consist of three concentric layers of cells; medulla, cortex
and cuticle of the hair.
Cuticle
The cuticle is the outermost layer made of flattened cells
arranged in an overlapping roof like pattern thickness of
each cell is 0.5 µm.
Cuticle is also responsible for surface properties of hair,
shine and smoothness.
Cortex
It forms the bulk of the hair and it consists of numerous
layer of flattened elongated cells packed together. It gives
strength elasticity to the hair. It also gives color to hair due
to presence of pigment.
Medulla
Medulla is the innermost layer of hair. The presence or
absence of medulla does not have too much to do with
human hair behavior, when washed with water, on color or
on curl it.
Dandruff (Wilkinson J. B et al., 1990, p-367)
Dandruff is a common condition among men and women of
all ages, and often causes skin irritation, embarrassment and
annoyance to sufferers. In consequence, it has been the
subject of numerous investigations and the target of many
forms of therapy. So far there has been little success in
explaining or curing the disease.
Definition: (Wilkinson J. B et al., 1990, p-17)
The disorder of the skin characterized by massive flaking of
the stratum corneum which continues to curiosity of
cosmetic chemist, is dandruff. No clinical analogy exists for
this condition and views as to its etiology come from many
areas of interest.
The disease is simply a microbial infection with the yeast
Pityrosporum ovale. This lipophilic organism which resides
on the stratum corneum of the human scalp has been studied
extensively without conclusive evidence for its involvement
in dandruff being forthcoming.
The theory depends upon the secretion by the organism of a
metabolite or waste products which enters the skin and
invokes large scale flaking from the stratum corneum.
Direct contact of the skin with the products of metabolism
of the organism has failed to give conformation of this
action. However, the treatments for dandruff offered at
present do have an ability to reduce the vitality of the
organism, but it is not clear whether this is their primary
function.
Types of Dandruff: (Virtue’s Family Physician, 1981)
Dandruff can be classified as disorders of the sebaceous
gland or skin scaling disorders. Dandruff can be of two
types,
Oily dandruff
Dry dandruff
Dry dandruff again can be mild or severe.
Oily dandruff (Pityriasis Steatoides):
On the scalp, waxy, greasy, yellowish, thick scales crusts
are present. Beneath the crusts, the scalp is red or pale but
dry. The hair may be dull and flat without shine. There may
be slight itching. If irritated, eczematization complicates the
condition to produce seborrhoeic dermatitis. Patients with
pityriasis steatoides usually develop thing and later loss of
hair. (Puri I, 1979).
Dry dandruff (Pityriasis sicca):
The scales are fine, thin, furfuraceous, white or grayish and
dry or only slightly greasy. The hair is dry and lusterless.
There is mild to moderate itching. The scales fall freely on
the shoulders. This type of dandruff is more common in
winter than in summer. It signifies exaggeration of normal
exfoliation of the horny layer of the epidermis. It usually
affects people with dry integument and scalp. In nutritional
disorders, scaliness of the scalp is exaggerated. (Behl P. N,
1982).
Etiology of dandruff:
The causation of dandruff is still debatable and it seems
undoubtedly that it is the sum total of many factors working
simultaneously.
The discussion about the cause of dandruff revolves around
the relative roles of physiological traumatic and infective
factors. (Wilkinson J. B et al., 1990, p-418 and Mitsui T,
1997)
Some of the causes of dandruff are:
• Abnormal keratinization of the epidermal
tissue
• Excessive lipid secretion
• Abnormal proliferation of scalp bacteria
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The yeast like organism, Pityrosporum ovale and
Pityrosporum orbicular are common members of the scalp
flora. Pityrosporum ovale was first described in 1874 by
Malassez, who believed it to be the role cause of dandruff.
(Wilkinson J. B et al., 1990, p-158)
Pityrosporum or Melassezia: (Woods G. L., Gutierrez Y,
1993)
The genus Melassezia contains at least two species of
globose to ellipsoid, unipolar, budding yeasts i.e.
Pityrosporum ovale and Pityrosporum orbicular. The
organisms bud repeatedly forms a broad base from some
location, forming a ‘collarette’ from which younger bud
emerge.
The pathogenesis of infections caused by Malassezia furfur
is not completely understood.
Other causes of dandruff: (Butler H, 2000)
! External factors:
Biochemical changes of the cutaneous scalp.
Increase in the number and activity of bacteria and fungi.
Localized inflammatory reaction following the use of
topical medications and cosmetics.
! Internal factors:
Hormonal imbalance.
Impaired metabolic nutrition.
Dietary factors.
Nervous tension.
SHAMPOO
Originally, shampoos were made up of soap or mixtures of
soaps; today synthetic detergents are used in the majority of
commercial products. (Balsam M. S., Sagarin E, 2008, p-
73).
Definition
Shampoo is defined as “a preparation of a surfactant (i.e.
surface-active material) in suitable form-liquid, solid, or
powder- which when used under the condition specified will
remove surface grease, dirt, and skin debris from the hair
shaft and scalp without affecting adversely the hair, scalp,
or health of user. (Balsam M. S., Sagarin E, 2008,p-74)
In earlier, a shampoo was defined as ‘a suitable detergent
for the washing of hair, packaged in a form convenient for
use’. The Good housekeeping consumer panel found that
women wanted a shampoo ‘to clean and also to rinse out
easily, impart gloss to the hair and leave it manageable and
non-drying’. (Wilkinson J. B., Moore R. J, 1990, p-398)
Ideal properties of shampoo (Sharma P. P, 1994)
• It should remove soil, sebum and residues
of hair setting lotions or dressing from
hair and scalp.
• It should produce the foam of the degree
that will satisfy the user.
• It should be easy to remove shampoo by
rinsing leaving the hair soft, lustrous and
manageable.
• It should impart a pleasant fragrance
during use, masking the odour of wet hair.
• Ease of combing of the wet hair. This
evaluates the roughness of the hair
immediately after treatment with the
detergent, and under conditions where this
roughness is most apparent. The consumer
associates this property with the cleansing
action of the shampoo.
• It goes without saying that the shampoo
detergent must be safe for use on the
scalp, and no irritation reddening or other
discomfort should be caused by its use.
INTRODUCTION: Piroctone olamine (O’Neil M. J et al., 2006)
Piroctone olamine acts as anti-microbial agent. It is used as anti-microbial against various microorganisms including bacteria,
fungi, mould, yeast etc. With reference to literature survey, in this present project work, an attempt has been made to evaluate the
anti-fungal property of Piroctone olamine against Pityrosporum ovale, Candida albicans and Malassezia furfur. This is the main
reason to take up this study
INCI Name : Piroctone olamine
Common Name: Piroctone olamine
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Chemical Name: 1-Hydroxy-4-methyl-6-(2, 4, 4-trimethylpentyl)
2-(1H) pyridinone, 2-aminoethanol salt
Empirical formula:
Emp. Formula : C14H23NO2 C2H7NO
Mol. Weight : 317.7 g/mol
Structural Formula:
Physical-chemical properties:
Appearance : White or slightly yellow crystalline powder.
Solubility : Freely soluble in 10% ethanol in water; soluble in
solution containing Surfactant in water or in 1%- 10%
ethanol; slightly soluble in water and in oil.
PH : 8.5-10.0 (1% suspension in water, 20°C)
Melting range : 130-135°C
EXPERIMENTAL ANALYSIS OF PIROCTONE
OLAMINE (ACTIVE):
Physical And chemical analysis:
The physical and chemical analysis of Piroctone olamine
was carried out as directed in analytical report of Piroctone
olamine.
1) pH (The Indian Pharmacopeia,2010, p-146):
Apparatus: Beaker, pH meter, stirrer, wash bottle.
Procedure: The pH meter was calibrated by using
buffer solutions. 1% solution was prepared in distilled
water and pH was determined by using pH meter.
2) Melting Point (The Indian Pharmacopeia,2010,
p-140):
Apparatus: Beaker, water bath, capillary tube,
thermometer, theils tube
Procedure: As per mentioned in Indian Pharmacopeia,
procedure was followed and melting point was
determined.
3) Solubility (The Indian Pharmacopeia,2010, p-
147):
Apparatus: Test tube, beaker
Procedure: As per mentioned in Indian Pharmacopeia,
procedure was followed and solubility was determined.
4) Residue on Ignition (The Indian Pharmacopeia, 2010, p-87):
Apparatus: Silica crucible, sulfuric acid, muffle furnace, weighing balance. Procedure: As per mentioned in Indian
Pharmacopeia, procedure was followed and residue on ignition was determined.
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Observation:
Table no. 1- Observation for Residue on Ignition:
Sr. No. Materials to be weighed Wt. (in gm) Wt. of residue after each heating (in gm)
1 Wt. of empty silica crucible 13.534 14.040
2 Wt. of silica crucible + sample 16.608 13.682
3 Wt. of sample taken 3.074 13.536
Calculation:
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5) % Purity: (as mentioned in standard testing procedure of Yasham Bio-science, Mumbai)
Apparatus: Beaker, conical flask, volumetric flask, burette, stirrer.
Procedure: 0.3 g of Piroctone olamine was weighed and dissolved in 50 ml of ethanol and 20 ml water then titrated with 0.1
M sodium hydroxide, by using phenol red solution as indicator, until a reddish violet color was obtained.
Then by using “1 ml of 0.1 M NaOH is equivalent to 0.01381 g of Piroctone olamine” this factor, the percentage purity was
determined.
Observation:
Table no. 2- Observation for Percentage Purity:
Sr. No. Materials to be weighed Wt. (in gm)
1 Wt. of butter paper 0.759
2 Wt. of butter paper + sample 1.084
3 Wt. of sample taken 0.311
Burette reading (B.R.) = 23.1 ml
Calculation:
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Where,
B.R. – Burette reading
N – Normality of NaOH
W – Weight sample taken
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ANTIMICROBIAL ANALYSIS OF PIROCTONE
OLAMINE: (Dubey R. C., Maheshwari D. K.; Practical
Microbiology, 2005, p-154, 155)
The active ingredient Piroctone olamine was subjected to in-
vitro testing by using agar-diffusion method to evaluate its
anti-fungal activity against Candida albicans, Malassezia
furfur and Pityrosporum ovale.
The cultures of above organisms were procured from Rajiv
Gandhi Institute of Bio-Technology, Nagpur. The
cultures were sub-cultured and grown in suitable growth
media.
Preparation of media:
For Candida albicans and Pityrosporum ovale:
The culture of Candida albicans and Pityrosporum ovale
were cultivated and maintained on a growth medium
described below. The incubation was done at room
temperature for 48 hours.
Potato dextrose agar (PDA):
39 gm of PDA and 5 gm agar-agar were weighed and
transferred to 1000 ml borosil beaker to this 1000 ml of
distilled water was added and was stirred on water bath for
20 minutes until frothing appears during heating. Then
sterilization was done in autoclave at 121 ˚C and 1.05
kgf/cm2 pressure for 20 minutes. After sterilization it was
kept in refrigerator.
For Malassezia furfur:
The culture of Malassezia furfur was cultivated and
maintained on a growth medium described below. The
incubation was done at room temperature for 7 days.
Sabouraud dextrose agar (SDA) with corn oil:
30 gm of SDA and 5 gm agar-agar were weighed and
transferred to 1000 ml borosil beaker to this 1000 ml of
distilled water was added and was stirred on water bath for
20 minutes until frothing appears during heating.
Preparation of solution of Active ingredient:
1 gm of sample which is to be analyzed was weighed to
make 1% solution in sterile distilled water.
Media used for screening:
For Candida albicans and Pityrospum ovale, Potato
Dextrose Agar was used. For Malassezia furfur Sabouraud
Dextrose Agar with corn oil was used
Procedure:
1. All the apparatus and media were sterilized in an
autoclave at 121 ˚C and 1.05 kg/cm2 pressure for 20
minutes.
2. The sterile and melted media were cooled at 45
˚C, then poured in labeled Petri plates and allowed to
solidify.
3. After solidification of media, the pure culture of
Candida albicans, Malassezia furfur and Pityrosporum
ovale were spread on the solidified media for respective
species.
4. The wells were made in each labeled plate with
the help of sterile cork borer.
5. Previously prepared 1% sample solution of
active was poured in different labeled Petri plates. For each
species one plate was kept as positive control, in which only
distilled water was poured.
6. Then all plates were incubated at 37 ˚C for 48
hours
7. After incubation, the zones of inhibition were
measured in mm of each species.
RESULT AND DISCUSSION: Common results of analysis Piroctone olamine:
Table no. 3- Analytical test observation for Piroctone olamine:
Sr.
No. Test Specification Results Inference
1 Colour White to Creamish Complies Passes test
2 Odour To match Std. Complies Passes test
3 pH (1% Solution) 8.5 - 10.0 9.48 Passes test
4 Melting Point 130 - 136 ˚C 133.7 ˚C Passes test
5 Solubility Freely soluble in 10% ethanol, Slightly soluble in
water, completely soluble in surfactant solution Clear Solution Passes test
6 Residue on
Ignition (%) Max. 0.1 0.065% Passes test
7 Purity (%) 98.0 – 101.5% 99.49% Passes test
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Anti microbial activity observation:
The zone of inhibition of Piroctone olamine against Candida albicans, Malassezia furfur and Pityrosporum ovale were measured
and are given in table no.4
Table no. 4- Antimicrobial test observation for zone of inhibition (in mm):
Sr.
No. Microorganism Concentration of Active
0% (Blank) 1%
1 Candida albicans (Fig 1,2,3,4) 00 mm 27 mm
2 Malassezia furfur 00 mm 20 mm
3 Pityrosporum ovale 00 mm 24 mm
Zones of Inhibition of Active against Candida albicans:
Before Incubation After Incubation
Fig 1: Distilled Water (Blank)
Fig 2: Distilled Water (Blank)
Fig 3: 1% Active Solution
Fig 4: 1% Active Solution
Zones of Inhibition of Active against Malassezia furfur:
Before Incubation After Incubation
Fig 5: Distilled Water (Blank)
Fig 6: Distilled Water (Blank)
Fig 7: 1% Active Solution
Fig 8: 1% Active Solution
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Zones of Inhibition of Active against Pityrosporum ovale:
Before Incubation After Incubation
Fig 9: Distilled Water (Blank)
Fig 10: Distilled Water (Blank)
Fig 11: 1% Active Solution
Fig 12: 1% Active Solution
CONCLUSION: Every individual wishes healthy hair on the scalp which plays a very important role in the social and personal
life. From this study we could understand the structure of hair, the types of dandruff and the types of shampoo. It is very
important to understand the type of hair one possess and respectively the choosing of the shampoo. Here is a small attempt to
understand the same which is helpful for everyone to maintain the hairs on the scalp in a better way. From above observation it
was concluded that, Piroctone olamine has excellent anti-dandruff activity against above dandruff causing species which can be
used in the anti dandruff shampoo formulation.
REFERENCES:
Balsam M. S., Sagarin E.(2008). Cosmetic Science
and Technology. 2nd
Edition. 2:73
Balsam M. S., Sagarin E.(2008). Cosmetic Science
and Technology. 2nd
Edition. 2:74
Behl P. N. (1982). Practice of Dermatology. 5th
Edition. Pg. No. 419
Butler H., Poucher’s (2000). Perfume, Cosmetics
& Soap; 10th
Edition. Kluwer Academic
Publishers, London; Pg. No.580
Dubey R. C., Maheshwari D. K. (2005) Practical
Microbiology. 1st edition. S. Chand and Company
Ltd.; Pg. No. 154,155
Government of India, Ministry of Health and
Family Welfare. Indian Pharmacopeia, 2010; Vol.
1. The Indian Pharmacopeia Commission,
Ghaziabad; Pg. No. 82
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www.jobb.co.in International, Peer reviewed, Open access, Bimonthly Online Journal
!
Government of India, Ministry of Health and
Family Welfare. Indian Pharmacopeia, 2010. Vol.
1.The Indian Pharmacopeia Commission,
Ghaziabad; Pg. No. 140
Government of India, Ministry of Health and
Family Welfare. Indian Pharmacopeia, 2010. Vol.
1. The Indian Pharmacopeia Commission,
Ghaziabad; Pg. No. 146
Government of India, Ministry of Health and
Family Welfare. Indian Pharmacopeia, 2010; Vol.
1. The Indian Pharmacopeia Commission,
Ghaziabad; Pg. No. 147
Mitsui T. (1997). New Cosmetic Science. 1st
Edition. Elseiver Publication. Pg. No. 158
O’Neil M. J., Heckelman P. E., Koch C. B., Roman
K. J. (2006). The Merck Index. 14th
edition. Merck
& Co., Inc., Whitehouse Station, NJ, USA.
Pg. No. 1293
Phate R. P. (1994). Book of Anatomy Physiology
and Health Education. 3rd
Edition.Career
Publication. Pg. No. 244
Phate R. P. (1994). Book of Anatomy Physiology
and Health Education. 3rd
Edition.Career
Publication. Pg. No. 246
Puri I. (1979). Beauty and Skin Care; 1st Edition.
Birla Publishing Pvt. Ltd. Pg. No. 50
Sharma P. P. (1994). Cosmetic Formulation,
Manufacturing & Quality Control. 4th Edition.
Vandana Publications Pvt. Ltd., Delhi; Pg. No. 303
Tortora G. J., Derrickson B. H. (2009). Principles
of Anatomy and Physiology; 12th
Edition.
Universal Publishing Corporation.1:55
Tortora G. J., Derrickson B. H. (2009). Principles
of Anatomy and Physiology.12th
Edition. Universal
Publishing corporation. 1:157
Virtue’s Family Physician (1981). 1st edition.
Published by Virtue and Company limited. 3:116
Wilkinson J. B., Moore R. J. (1990). Harry’s
Cosmeticology. 7th
Edition. Chemical Publishing
Company INC. New York; Pg. No. 17
Wilkinson J. B., Moore R. J. (1990). Harry’s
Cosmeticology. 7th
Edition. Chemical Publishing
Company INC. New York; Pg. No. 158
Wilkinson J. B., Moore R. J. (1990). Harry’s
Cosmeticology. 7th
Edition. Chemical Publishing
Company INC. New York; Pg. No. 367
Wilkinson J. B., Moore R. J. (1990). Harry’s
Cosmeticology. 7th
Edition. Chemical Publishing
Company INC. New York; Pg. No. 378
Wilkinson J. B., Moore R. J. (1990). Harry’s
Cosmeticology. 7th
Edition. Chemical Publishing
Company INC. New York; Pg. No. 418
Woods G. L., Gutierrez Y. (1993). Diagnostic
Pathology of Infectious Diseases; 1st Edition. Pg.
No. 418
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Source of Support: Nil Conflict of Interest: None Declared
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