Journal of Biotechnology and Biosafety ...

Journal of Biotechnology and Biosafety !

"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!

*778!(9((2565: J o u rna l o f B io techn o lo gy an d B iosa fety

!

www.jobb.co.in International, Peer reviewed, Open access, Bimonthly Online Journal

!

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:

[email protected]

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.


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

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)


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

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


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

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


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

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.


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

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:

!"#$%&"!!"!!"#$%$&#!!!! !!""!!!!!!! !!!!

!

!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!""!!!!"!!"#!! !!"!!"#!

!!!"#

!"#$%&"!!"!!"#$%$&#! ! ! !!!"#!

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:

!"#$%&!!!! !!!!!!!!!!!"#$"!!!""

!!!!!!

Where,

B.R. – Burette reading

N – Normality of NaOH

W – Weight sample taken

!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!"!!!!!!!"#!!!!"#$"!!!""

!!!!!!!!""

!! !!!!!!!!!!!!!!"##

!!!"##

!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!"#$%&! ! ! !!!!!"!


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

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


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

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 3: 1% Active Solution


Zones of Inhibition of Active against Malassezia furfur:







"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!

Zones of Inhibition of Active against Pityrosporum ovale:






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


"#$%&'!()!*++%'!,)!-./%.0123'40%.01!(5,6)!(,29,!


!


!


Family Welfare. Indian Pharmacopeia, 2010. Vol.

1.The Indian Pharmacopeia Commission,



Family Welfare. Indian Pharmacopeia, 2010. Vol.




Family Welfare. Indian Pharmacopeia, 2010; Vol.



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


Cosmeticology. 7th




Cosmeticology. 7th




Cosmeticology. 7th




Cosmeticology. 7th



Woods G. L., Gutierrez Y. (1993). Diagnostic

Pathology of Infectious Diseases; 1st Edition. Pg.

No. 418

http://tinyurl.com/cfbhs22

Source of Support: Nil Conflict of Interest: None Declared

Date post:	28-Jan-2022
Category:	Documents
Upload:	others
View:	3 times
Download:	0 times

Journal of Biotechnology and Biosafety ...

Documents