PREPARATION AND CHARACTERIZATION OF THE POLY HERBAL ANTI-AGEING CREAM

Presented by :-
Avinash V. More
B.tech.(Cosmetics)
Guided by :-
Prof. Dr. K. R. Biyani
M. Pharm., Ph.D., D.P.P.M.
Department of cosmetic technology
Anuradha college of pharmacy
Anuradha nagar, sakegaon road,chikhli. Dist- buldana (m.S.) 443201
PREPARATION AND CHARACTERIZATION OF
THE POLY HERBAL ANTI-AGING CREAM

INTRODUCTION
SKIN
 The skin is the largest organ of the body, with a total area of about 20 square
feet.
 Functions Of Skin
- The skin protects us from microbes and the elements.
- regulate body temperature.
- permits the sensations of touch, heat, and cold.
- Storage and synthesis
- Absorption
- Water resistance
- Excretion

STRUCTURE OF THE SKIN
The skin consists of three main layers:
-Epidermis (the outer layer)
-Dermis (the middle layer)
-Subcutaneous or Hypodermic

 LAYERS OF EPIDERMIS
i) Stratum Basale – It is the deepest layer of epidermis and has a single
layer of keratinocytes. It is also known as stratum germinativum.
ii) Stratum Spinosum – It is superficial to the stratum basale. It
provides both strength and flexibility to the skin. This layer consists
of 8‐10 layers of keratinocytes.
iii) Stratum Granulosum – It is middle layer of the epidermis. It
consists of protein called keratohyalin which converts tonofilaments
into keratin. This layer consists of 3‐5 layers of flattened
keratinocytes. Also present in the Keratinocytes are membrane
enclosed lamellar granules which release lipid rich secretion. This
secretion fills the space between cells of stratum granulosum, stratum
lucidum and stratum corneum. They act as a water repellent sealant
that helps retard loss of body fluids and entry of foreign materials.
iv) Stratum Lucidum – This layer is present only in those areas which
are prone to friction i.e. in thick skin. It consists of large amount of
keratin and thickened plasma membrane. The layer is made up of 3‐5
layers of flattened dead keratinocytes.

V) Stratum Corneum – This layer consists of 25‐30 layers of flattened
dead keratinocytes. These are continuously shed and replaced by cells
from the deeper strata. This layer consists of mostly keratin and lipid
secretions from lamellar granules which make this layer an effective
water repellent barrier. Multiple layers of dead cells protect the deeper
layers from injury and microbial invasion.
 DERMIS
Dermis is composed mainly of connective tissues containing
collagen and elastic fibers. Cells present in dermis include:‐
i) Fibroblast – They provide the structural framework for many tissues and play
a critical role in wound healing. They are also responsible for synthesizing the
dermal proteins.
ii) Macrophages – They are also called as big eaters as their role is to
phagocytose cellular debris and pathogens.
iii) Adipocytes – They are cells specialized in storing energy as fat.

 Based upon the tissue structure dermis is divided into 2 regions
i) Papillary region – It is a superficial part of the dermis. The surface area
of the papillary region is greatly increased by small fingerlike projection
called dermal papillae. Some dermal papillae contain tactile receptors called
corpuscles of touch or Meissner corpuscles these are nerve endings that are
sensitive to touch. Also present in dermal papillae are free nerve endings
which initiate signal which gives rise to sensation of warmth, coolness,
pain, tickling and itching.
ii) Reticular region – It is the deeper part of dermis. In this region bundles
of collagen fibers are interlaced in a net like manner. Adipose cells, hair
follicles, nerves, sebaceous glands, sweat glands occupy the space between
the fibers. Combination of collagen and elastic fibers in reticular region is
responsible for providing the skin with strength, extensibility and elasticity.

 HYPODERMIS
- It is also called as subcutanous tissue.
- Deepest layer of the skin located under the dermis.
- Consisting a network of collagen and fat cells
- Acts as a protecting underlying tissues from cold & trauma.
- Made up of loose connective tissue and fat.
- Play important in the regulation of temperature of skin and body.

 SKIN AGEING
 Skin ageing is the result of continual deterioration process because of
damage of cellular DNA and protein.
 Ageing process is classified into two distinct type, i.e. “sequential skin
ageing” and “photo-ageing”.
 sequential skin ageing is universal and predictable process characterized by
physiological alteration in skin function. In the ageing process
keratinocytes are unable to form a functional stratum corneum and rate of
formation from neutral lipids slows down, resulting in dry pale skin with
wrinkle.
 In contrast, photo ageing is caused by over exposure to uv rays from
sunlight. It is characterized by dry, pale and shallow skin, displaying fine
wrinkles as well as deep furrows caused by the disorganization of epidermal
and dermal components associated with elastosis and heliodermatitis6.
 Skin ageing is influenced by many factors including ultraviolet radiation
(UV), excess alcohol consumption, tobacco abuse and environmental
pollution. Combined, these factors lead to cumulative deterioration in skin
appearance and function.

 Ageing of the skin is primarily related to reductions in the levels of Type I
collagen, which is the principal component of skin dermis.
 Type I collagen is the main structural component of the extracellular matrix
(ECM), which is known to perform a pivotal function in the maintenance of
the structure of the skin dermis.
 Several molecules have been reported to increase type I collagen synthesis,
namely, transforming growth factor-β (TGF-β) and fibroblasts growth factor
2 (FGF2) to prevent the skin from ageing or wrinkle.
 Plant extracts rich in phytochemicals, like flavonoids, phenolic acids,
saponins and alkaloids, having collagen synthesis activity, are being widely
used for the development of anti-skin-ageing topical cosmetic products.

 Anti-ageing
 Antiageing substances are intended to prevent or limit the process
of becoming old. There are two main groups of agents that can be used
as anti-ageing cream components, the antioxidants and the cell
regulators.
 The antioxidants, such as vitamins, polyphenols and flavonoids, reduce
collagen.
 The cell regulators, such as retinols, peptides and growth factors (GF),
have direct effects on collagen metabolism and influence collagen
production.

COSMETIC FOR SKIN
 Cosmetics, also known as make-up, are substances or products used to
enhance the appearance or fragrance of the body. Many cosmetics are
designed for use of applying to the face and hair.
 In the 21st century, women generally use more cosmetics than men. They
are generally mixtures of chemical compounds; some being derived from
natural sources (such as coconut oil), and some being synthetics. Common
cosmetics include lipstick, mascara, eye shadow, foundation, rouge, skin
cleansers and skin lotions, shampoo, hairstyling products (gel, hair spray,
etc.), perfume and cologne.
 In the U.S., the Food and Drug Administration (FDA), which regulates
cosmetics, defines cosmetics as "intended to be applied to the human body
for cleansing, beautifying, promoting attractiveness, or altering the
appearance without affecting the body's structure or functions". This broad
definition includes any material intended for use as a component of a
cosmetic product. The FDA specifically excludes soap from this category.

HERBAL COSMETICS
 Herbal cosmetics also known as natural cosmetics or Ayurvedic cosmetics .
With the beginning of the civilization, mankind had the magnetic dip
towards impressing others with their looks.
 At the time, there were no fancy fairness creams or any cosmetic surgeries.
The only thing they had was the knowledge of nature, compiled in the
Ayurveda.
 With the science of Ayurveda, several herbs and floras were used to make
Ayurvedic cosmetics that really worked.
 Ayurvedic cosmetics not only beautified the skin but acted as the shield
against any kind of external affects for the body.

 The best thing of the herbal cosmetics is that
- it is purely made by the herbs and shrubs.
- The natural content in the herbs does not have any side effects on the human
body; instead enrich the body with nutrients and other useful minerals.
- Cosmetics are the products in which herbs are used in crude or extract form.
The basic idea of skin care cosmetic lies deep in the Rigveda, Yajurveda,
Ayurveda, Unani, Homeopathic system of medicine.
- In this modern era, knowledge of usage of herbs is being blend with
advanced cosmetic technology to develop a safe, elegant beauty product,
which has wider range of people acceptability.
- Basically it is beauty invented by nature & perfected via technology
- Polyherbal formulations are now a day’s one of the new delivery system in
India. Using this concept I will be going to prepared the polyherbal
formulation for anti ageing cream.

BENEFIT
 Improves complexion
 Smoothens wrinkles
 Moisturizes and hydrates ageing and sun-damaged skin
 Neutralizes environmental damage
 Anti Aging Cream keeps wrinkles at bay and adds a natural glow to your
skin, and reduces fine lines
 Rich with natural oils,herbs and fruit extracts, the cream works gently on
the skin.
 The cream visibly reduces wrinkles and neutralizes environmental damages
to give back that youthful suppleness.

REVIEW OF LITERATURE
 Y. H. Kim, K. H. Kim; et al (2010)
Skin aging is characterized by a progressive deterioration of the
skin'sfunctional properties, linked to alterations of dermal connective tissue
due to the changes at the cell, gene and protein levels. Skin aging can be
divided into two basic processes: intrinsic aging and photoaging .
 F. Perin ; et al (2016)
Cutaneous ageing cab be defined as the result of two different and
cumulated processes: intrinsic and extrinsic ageing (also known as
photoageing). While intrinsic ageing is natural and mainly due to the
passage of time (influence of genetic factors, oxidative stress, cellular
senescence etc.) and its consequences, photoageing is mainly linked with
the detrimental effects of solar exposure on the skin, although pollution, diet
and smoking are also contributing factors.

 S. Rao, F. Muia; et al (2013)
Skin barrier function, principally the stratum corneum, is the primary
line of defence against extrinsic stress such as UV‐induced photo‐damage,
microbial infections and physical deterioration resulting from ageing and
environmental exposure. Scientific evidence suggests that both intrinsec and
invornmental factors contribute to „compromised” skin barrier function.
The stratum corneum functions as an effective barrier and is critical for
controlling and preventing water loss.
 P.Boonme ; et al (2011)
Application of cosmetic products containing oils with antioxidant
activity is widely acceptable to benefit healthy skin.
 N. J.Lowe; et al (2013)
Aging affects all levels of the skin. From the stratum corneum
downwords aging creates corneocyte dysfunction, epidermal atrophy,
dysplasia and abnormal pigmentation.

 D. Whitby, J. Allen; et al (2012)
Aging occurs in all organs of the body; however, the skin appearance,
such as wrinkles and furrows, is markedly observed for aging notices. It is a
challenging work for cosmetic scientists to find the means for reducing the
changes on the skin appearance due to aging. Wrinkles appear over time due
to changes in the support structures of the skin from chronological ageing,
but photoageing speeds the process considerably leading to quickly formed,
deep wrinkles.
 M. Paye, A. Barel; et al (2001)
Skincare products that affect wrinkles are a reality and are well
established in consumer, practitioner and corporate perspectives. In the
broadest definition, ‘‘products’’ range from classic and simple cosmetic
preparations through vitamins, antioxidants, topical and oral cosmeceutical
and pharmaceutical preparations, and even to surgical and laser
interventions.

 P. Boonme; et al (2011)
There are a lot of treatments boasting the capability to improve
wrinkles: pharmaceutical, surgical and cosmetic solutions. These treatments
are intended to change the nature of ageing collagen, stretch the skin, fill in
the depressions of the skin or paralyse the muscles that cause the wrinkle.
Retinoid products, for example, act by inhibiting enzymes from breaking
down collagen, but they may produce redness, burning and general
discomfort .
 L. Baumann; et al (2016)
Regardless of the etiology of skin aging, there are important
characteristics of aged skin that must be considered. These changes occur
throughout the epidermis, dermis, and subcutaneous tissue and can result in
wide‐ranging alterations in the topography of the skin.

 Sparavigna, B. Tenconi; et al (2013)
alpha‐hydroxy acids penetrate into the top layer of the skin,
producing only subtle improvement, though, and causing a mild and
temporary irritation, increasing the skin’s sensitivity to the sun and
particularly increasing the possibility of sunburn .
 C. Rona, F. Vailati; et al (2004)
Wrinkle cosmetic treatment is exerted by a variety of active
functional ingredients: modern anti‐ageing cosmetics go well beyond the
simple moisturizing function of traditional anti‐wrinkle creams, by exerting
a more complex function in protecting the skin from external injuries,
nourishing it, and removing its superficial layers. That is the reason why the
term “cosmetceuticals” is increasingly used: there are many cosmetic
products that fit into this category defined in the regulatory systems of some
countries .

 Aburjai T, Natsheh M.; et al (2003)
Prunus amygdalus, commercially known as almonds is a natural
product whose seeds are rich in polyphenolic compounds especially
flavonoids and phenolic acids. Almond oil has excellent emollient properties
help the skin to balance water loss and absorption of moisture, helps relieve
irritation,inflammation and itching, and is greatly lubricating.
 Sharma SK; et al. (2006)
Reported that Oxygen free radicals induce damage due to
peroxidation to bio membranes and also to DNA, which causes tissue
damage. Antioxidants neutralise the effect of free radicals through different
ways and may prevent the body from various diseases. Synthetic
antioxidants such as butylated hydroxytoluene (BHT) and butylated
hydroxyanisole (BHA) have recently been reported to be dangerous for
human health. Thus, the search for effective, non-toxic natural compounds
with antioxidative activity has been intensified in recent years. So they
included a brief account of research reports on plants with antioxidant
potential.

 Priya OS; et al (2013)
Plants are the main source of natural antioxidants in the form of
phenolic compounds (phenolic acids, flavonoids and polyphenols). Most of
the anti-inflammatory, digestive, antinecrotic, neuroprotective and
hepatoprotective drugs derived from natural origin have been reported to
have antioxidant/radical scavenging mechanism as part of their activity.
 M.S. Ashawat ; et al (2009)
The plant parts used in cosmetic preparation should have varieties of
properties like antioxidant, anti-inflammatory, antiseptic, emollient,
antiaging, antiseborrhatic, antikerolytic activity and antibacterial etc. Herbal
products claim to have less side effects, commonly seen with products
containing synthetic agents. The market research shows upward trend in the
herbal trade with the herbal cosmetic industry playing a major role in
fueling this worldwide demand for herbals.

AIM & OBJECTIVE
 Aim
Preparation and Characterization of the Poly Herbal Anti-ageing
Cream.
 Objective
- Formulation of the poly herbal anti-ageing cream using the extracts.
- Preparation of extract with different solvent of leaves of two different
identified plants.
- Priliminary phytochemical investigation of different extract of the plants.
- To prevent aging of skin
- To reduce the rate of premature ageing
- To nourish and beautify the skin .

NEED OF WORK
 Natural remedies are more acceptable in the belief that they are safer with
fewer side effects than the synthetic ones.
 Herbal formulations have growing demand in the world market.
 To prepare and evaluate the polyherbal cosmetic cream comprising extracts
of natural products such as aloe, orange peel extract, punica extract and
green tea.
 The development and evaluation of the poly herbal cream containing hydro-
alcoholic extract of aloe (Aloe vera) and citrus aurintum extract Although
various topical herbal extracts of Green tea (Camelia sinensis) and
pomegranate (Punica granatum) The plants have been reported good anti-
microbial, anti-oxidant and also reported as a good antiageing herbs.

PLAN OF WORK
 literature survey
 Procurement of Raw materials and Active
 Selection of base formulation
 Incorporation of active into base formulation & their optimization
 Development and optimization of final formulation.
 Evaluation of cream
 Stability study

Sr No Material Name Supplier
1 Stearic acid Anuradha college of pharmacy
2 Cetyl alcohol Anuradha college of pharmacy
3 Almond oil Anuradha college of pharmacy
4 Triethanolamine Anuradha college of pharmacy
5 Potassium Hydroxide Anuradha college of pharmacy
6 Glycerol Anuradha college of pharmacy
7 Methyl paraban Anuradha college of pharmacy
8 Propyl paraben Anuradha college of pharmacy
9 Water qs. 100 Anuradha college of pharmacy
10 Camellia sinesis extract Prepared in laboratory of Anuradha
college of pharmacy
11 Punica extract Prepared in laboratory of Anuradha
college of pharmacy
12 Citrus aurantium extract Prepared in laboratory of Anuradha
college of pharmacy
13 Aloe extract Prepared in laboratory of Anuradha
college of pharmacy
MATERIALS AND METHODS
Materials:-

Methods
- Heating
- Mixing
- Emulsification
- Packaging
- labeling

INGREDIENTS PROFILE
 Active profile
- Punica Granatum
- Orange Extract
- Aloe Vera
- Camelila Sinesis
- Almond Oil
 Excipients Profile
- Stearic Acid
- Cetyl Alcohol
- Glycerin
- Triethanolamine
- Methyl Paraben
- Propyl Paraben
- Distilled Water

 ACTIVE PROFILE
 Punica Granatum
Punica granatum Linn , Punicaceae family
 Synonyms - Granatum punica St Lag, Punica florida Salisb
 Chemical Constituents –
- Alkaloids: Punicalin, punicalagine, granatine β, gallagyldilaclene casuarine,
pedunculagine and tellimagrandine1 were isolated from the pericarp the fruits also
contain punicalagine, punicaline and grantine B. The bark contains isopelletierine,
pseudo- pelletierine, methyl isopelletierine and pelletierine.
- Tannins: Gallic acid, granatine A, corilagine and ellagic acid have been isolated
from the pericarp the fruit contains an ellagitanin and ellagic acid.
- Triterpenes and phytosterole: Sitosterol, friedelin, ursolic acid, mastinic acid asiatic
acid are present.
 Uses – polyphenols (such as catechins, gallic and ellagic acids) and punicalins and
punicalagins act as an wonderful antioxidant and free radical neutralizing
ingredients.

 Orange Extract
 Synonym – orange cortex
 Biological source :- The orange peels are dried or fresh outer part of
pericarp of ripe or nearly ripe fruits of Citrus aurantium Linn Family –
Rutaceae
 Description:-
Colour - Dark orange red
Odour - Aromatic
Taste - Aromatic and bitter
 Chemical constituents :- Bitter orange peels contain about 2.5 % of
volatile oil. It also contain several other components like Hesperidin , iso-
hesperidin , neo-hesperidin , vitamin C and Pectin. The bitter substance
present in the drug are glycosidal compounds known as aurantiamarin and
aurantimaric acid. The volatile of drug contain about 90% limonene, chiefly
contain citral and citronerol.

 Uses in Cosmetics :-
- The extract of orange act as an Absorbent and Binder.
- It also act as in Emulsion stabilizer and Flavoring agent in the formulation.
- It also use for the purpose of Viscosity controlling agent.
- It Acts as an emollient lubricating the skin’s surface, which gives the skin a
soft and smooth appearance and fights wrinkles.
- Packed with vitamin C, orange extract is another powerful antioxidant
youth molecule that promotes natural collagen production, even skin
complexion, and skin immunity.

 Aloe Vera
 Synonym – Aloe
 Biological source – It is dried juice Aloe Barbadenis , Family- Liliaceae
 Chemical constituents – Major substances of Aloe vera.
Anthraquinones: aloe-emodinaloetic acid,
aloin, anthranol, barbaloin, isoberbaloin , emodinester of cinnamic acid.
- Saccharides: cellulose ,glucose ,mannose, aldopentose, glucomannan
acetylated glycomannan, galactogalacturan , glucogalactomannan
galactoglucoarabinomannan.
- Vitamins : B1,B2,B6, vit. C, carotene,choline,folic acid
Enzymes:amylase, carboxypeptidase, catalase, cyclooxidase, lipase, oxidase
-Low molecular weight substances : arachidonic acid, cholesterol, gibberellin
lectin-like substances , lignins, salicylic acid, sitosterol, steroids,
triglyceridesuric acid

 Uses –
- Polysaccharides act as moisturizers, hydrating the skin.
- Aloe is absorbed into the skin and stimulates the fibroblasts to replicate
themselves faster and it is these cells that produce the collagen and elastin
fibres, so the skin becomes more elastic and less wrinkled.
- Aloe also makes the surface of the skin smoother because of its cohesive
effect on the superficial flaking epidermal cells by sticking them together.
- It also possesses the ability to interfere with the enzyme that produces
melanin deposits in the skin, preventing the formation of 'liver spots' which
tend to form in ageing34.

 Camelila Sinesis
 Synonym : Green Tea
 Biological source- Dried leaves of the plant Camellia Sinesis.
Family- Theaceae
 An extract of the leaves of the plant Camellia Sinesis. Commonly referred
to as green tea extract.
 The functions of these ingredients include: antifungal agent; antimicrobial
agent; antioxidant; cosmetic astringent; skin protectant; skin-conditioning
agent – emollient; skin-conditioning agent, anti-ageing agent.
 Chemical constituents :-
The camellia sinesis leaf extract consist of Flavonols and flavonol
,glycosides, Polyphenolic acids and depsides Other polyphenols. It also
consist of Caffeine Theobromine, Amino acids ,Organic acids,
Monosaccharides , Polysaccharides Cellulose, Protein ,Lignin ,Lipids
,Chlorophyll and other pigments.

 Uses:-
- The extract of the leaves of C. sinensis. Use as Antifungal agent;
antimicrobial agent; antioxidant; anti ageing agent emollient.
- Research has established that topical application of green tea leaves or
extracts have many benefits for skin, including anti-aging properties. The
polyphenols in green tea possess potent antioxidant and skin-soothing
properties, and show significant promise for improving the appearance of
sun-damaged skin.

 Almond Oil
 Name: Almond Oil
 INCI Name: Prunus Amygdalus Dulcis (Sweet Almond) Oil.
 Botanical name: Prunus amygdalus dulcis
 Chemical constituents:
Almond oil is high in mono and polyunsaturated fatty acids, minerals
and glycosides. Fatty acids are necessary along with glycerol for the cell to
function normally. The oil also contains vitamins A, B1, B2, B6 with small
amounts of Vitamin E and D. Due to the presence of Vitamin E the oil has
antioxidant capability. Antioxidants protect vital cell structures by
neutralizing free radicals.

 Uses:-
- Almond Oil is a rich source of essential fatty acids (oleic, linoleic),
triglycerides, proteins, vitamins A, D and E, having a good spreadability
almond oil has a geriatric activity on wrinkled skin, reducing fine lines and
wrinkle.
- The most beneficial qualities of Oil for skin rejuvenation include increased
levels of collagen production deriving from the fact that it can penetrate the
skin more deeply, relief of damaged skin caused by sun exposure, treatment
of dry skin.
- Almond oil is gives excellent emollient properties, soothing and nourishing
the skin with a good spreadability and without leaving any greasy feeling

 EXCIPIENTS PROFILE
 Stearic Acid
 Ingredient name: stearic acid
 INCI name : stearic acid, octadecanoic acid, stearophanic acid, N-
octadecamoic acid
 Description:-
Sr. No. Characteristics Specification
1 Colour Whitish crystal or powder
2 Odour Mild
3 solubility Insoluble in water
4 Melting point 68.8 C
5 Boiling point 383
6 Flash point 196

 Application
- It provides binding and thickening properties on the products that makes the
products stick smoothly to the skin and have extended shelf life.
- stabilizer
- Opaciying agent
- Emulsifying agent
- Whitening agent
- Texure modifier

 Cetyl Alcohol
 Ingredient name: Cetyl alcohol
 INCI name : Cetyl alcohol
 Molecular formula :
 Molecular waight 242.44 g/mol
 Description:- Sr. No. Characteristics Specification
1 Colour Whitish to pale yellow
2 Odour Charecteristics
3 solubility Insoluble in water
4 Melting point 49.56 C
5 Acid value <1
6 Hydroxyl value 200-228
7 Saponification value <2
Applications: - Non gelling thickener
- Viscosity and consistency enhancer
- Emollient
- Moisturizer
- Foam booster

 Glycerin
 Ingredient name: glycerol, glycerin, glycerine, 1,2,3- propanetriol, glycyl
alcohol
 INCI name : glycerin( cosmetics grade)
 Chemical formula : C3H8O3
 Molecular waight : 92.094 g/mol
1 Colour Colourless transparent liquid
3 Specific gravity 1.2613 at 25 C
4 Residue on
ignition
0.001<0.005
Applications:- -Emollient
-Solvent
-Humactant
-Moisturizer

 Triethanolamine (TEA)
 Ingredient name: Triethanolamine (TEA)
 INCI name : Triethanolamine (TEA)
 Chemical formula : C6H15NO3
1 Colour Colourless transparent
liquid
3 Boiling point 335.40 °C
4 Solubility in water 149 g L−1 (at 20 °C)
Properties - pH adjuster, stabilizes emulsions, fragrances & preservatives,
foam stabilizer (has detergent properties and stabilizes other surfactants),
improves efficacy of preservatives by stabilizing the pH value.

 Methyl Paraben
 Ingredient name: methyl paraben , methyl 4-hydroxybenzoate
 INCI name : methyl paraben
 Molecular waight 152.15 g/mol
 Description :- Sr. No. Characteristics Specification
1 Colour White
3 Solubility Slightly soluble in water, freely soluble in
alcohol
4 Melting point 125-128 C
5 Boiling point 149 C
6 Residue on
ignition
Not more than 0.1% w/w
7 Purity assay 99.96 % W/w
Application:- Anti-microbial agent.,
Preservative.

 Propyl Paraben
 Ingredient name: propyl paraben , propyl 4-hydroxybenzoate
 INCI name : propyl paraben
1 Colour White
3 Solubility Slightly soluble in wate, freely
soluble in alcohol
4 Melting point 96 to 99 °C
6 Residue on ignition Not more than 0.1% w/w
7 Purity assay 99.96 % W/w
Application:- Anti-microbial agent, Preservative.

 Distilled Water
 Ingredient name: Water
 INCI name : Distilled water
 Chemical formula : H2O
 Description:-
Sr. No. Characteristics Specification
1 Appearance Clear colourless,
odourless liquid
2 Density 1 gm/ml
3 Ph 7.0 – 8.0
4 Boiling point 100 C
Application:- Solvent, Solubilizer , Volume maker

EXPERIMENTAL WORK
 PREPARATION OF EXTRACTS.
 Preparation of Punica leaf and Camelia sinensis extract
- Punica granatum leaves and camellia tea were collected locally from
surrounding area.
- Air dried leaves were grinded to a fine powder in a suitable grinder mixture.
Shade dried powder was extracted using soxhlet extractor with distilled
water, alcohol, hexane separately to get semisolid extract.
- The organic solvents were recovered by steam distillation.
- The extracts were then concentrated to dryness under reduced pressure and
controlled temperature, respectively and they were preserved in a
refrigerator.

 Preparation of Aloe vera extracts
- Hydro alcoholic mixture was prepared by mixing one liter each of analytical
grade Ethanol and distilled water.
- One kg whole leaves, unpeeled, of Aloe Vera was cut into thin slices by
knife and was put into a glass beaker.
- Hydro alcoholic mixture was added to it and macerated for 48 hours. Glass
beaker was sealed with aluminium foil and kept in the laboratory.
- The beaker was shaken for 10 minutes after every12 hours. Finally the
macerated material of plant was filtered through several layers of muslin
cloth for coarse filtration.
- The coarse filtrate was then filtered through a Whatman # 01 filter paper.
The filtrate so obtained was evaporated under reduced pressure at 40 ºC in a
Rotary vacuum evaporator.
- The reddish brown colored extract so obtained was collected in Stoppard
glass tubes and stored in freezer at 0°C.

 Preparation of Orange Peel Extracts
- Orange fruits were washed by distilled water then peeled and their edible
portions were carefully separated.
- The peels were air dried in a ventilated oven at 40°C for 48 h and ground to
a fine powder and passed through a 24-mesh sieve according to the method
described by Van-Acker et al..
- 100g powdered sample was extracted with either 800ml ethanol or methanol
or dichloromethane or acetone or hexane or ethyl acetate at room
temperature by Soxhelt extraction method for 6 h.
- The mixture filtered through a Whatman No. 2 filter paper for removal of
peel particles. The residue was re-extracted twice under the same condition
to ensure complete extraction.
- The extracts were filtered and evaporated to dryness under reduced pressure
at 60°C by a rotary evaporator.
- The extracts were placed in dark bottles and stored in refrigerator at 4°C
until use.

 PRELIMANARY PHYTOCHEMICAL INVESTIGATIONS OF
DIFFERENT EXTRACT
 Detection Of Alkaloids
 Detection Of Aminoacids
 Detection Of Terpenoids
 Detection Of Phenolic Compounds And Tannins
 Test For Saponin Glycosides
 Test For Flavonoids

 PREPARATION OF CREAM BASE
Sr. No. Ingredients BF1 BF2 BF3 BF4
Phase A:- Oil Phase
1 Stearic acid 20 18 13 15
2 Cetyl alcohol 0.50 4 2 4
4 Almond oil 3 3 3 4
Phase B:- Aqueous phase
3 Triethanolamine 1.20 1.1 0.8 1.0
5 Potassium Hydroxide 0.36 0.36 0.36 0.50
6 Glycerine 8 8 5 6
7 Methyl paraban 0.2 0.2 0.2 0.2
8 Propyl paraben 0.2 0.2 0.2 0.2
9 Water qs. 100 Upto 100 ml Upto 100
ml
Upto 100
ml
Upto 100
ml

Procedure
- Oil in water (O/W) emulsion-based cream (semisolid formulation) was
formulated.
- The emulsifier (stearic acid) and other oil soluble components (Cetyl
alcohol, almond oil) were dissolved in the oil phase (Part A) and heated to
75°C.
- The preservatives and other water soluble components (Methyl paraban,
Propyl paraban, Triethanolamine, Propylene glycol) were dissolved in the
aqueous phase (Part B) and heated to 75° C.
- After heating, the aqueous phase was added in portions to the oil phase with
continuous stirring until cooling of emulsifier took place.

OPTIMIZATION OF BASE FORMULATION
 Determination of physical parameters of prepared base
 Determination of pH
 Determination of viscosity
 Spreadability test
 Thermal Stability Study

 OPTIMIZATION OF BASE FORMULATION
Sr. No. Parameters BF1 BF2 BF3 BF4
1 Appearance ++ + ++ +++
2 Consistency ++ + +++ +++
3 Odour ++ + ++ ++
Abbreviations: + Poor, ++ Good , +++ Better
Discussion:- From the above table it was found that formulation, BF4 does
not occur any changes in an appearance, consistency and Odour.
 Determination of physical parameters of prepared base
Procedure: Physical parameters like appearance , consistency , odour of each base
formulation was observed and note down as + Poor, ++ Good , +++ Better for to
choose most stable base formulation.

 Determination of pH
Apperatus : beaker , pH meter, stirrer, wash bottle.
Procedure: The pH meter was calibrated using a standard buffer sol. About
0.5 g of the cream was weighted and dissolved in 50 ml of distilled water
and its pH was measured at 27oC
Sr No Days
Formulation
BF1 BF2 BF3 BF4
1 Initial Day 6.4 6.2 6.2 6.3
2 7 days 6.2 6.3 6.5 6.2
3 15 days 6.3 6.4 6.5 6.2
4 21 days 6.3 6.3 6.5 6.3
5 30 days 6.2 6.4 6.5 6.2
Discussion:-From the observation the formulation BF1, BF2, BF3, BF4,
and all are shown desired pH (desired pH should be 5.5 to 7.5)

 Determination of viscosity
Appararus :-Brookfield viscometer ,beaker,thermometer and wash bottle
Procedure :- A 100 gm of each formulation was weighed and transpired to
beaker and the viscosity of formulation was determined with the help of
Brookfield Viscometer (DV II+ Pro model) using spindle number S64 at a
20 rpm at a temperature of 25o C.
Sr No days
Formulation
BF1 BF2 BF3 BF4
1 Initial day 20130 cps 22500 cps 23520 cps 25590 cps
2 7 days 21520 cps 23580 cps 22580 cps 25582 cps
Discussion: From the above observation formulation BF4 had most stable and
appropriate viscosity, than BF1,BF2,BF3.

 Spreadability test
Parameters
Formulation
BF1 BF2 BF3 BF4
Spreadability 26.33 ± 0.3 24.47 ± 0.4 22.35 ± 0.5 21.83 ± 0.6
Discussion
From the above observation, the formulation BF4 showed desired Spreadability
than BF1, BF2, and BF3.
 Determination of Thermal Stability:
Parameter Base Formulation
BF1 BF2 BF3 BF4
Thermal
Stability
No Oil Separation Oil Separation No Oil
Separation
No Oil
Separation
Discussion:
From above the formulation BF1, BF3, BF4, showed No oil separation but BF2
show oil separation.

 Stability Study
Formulation
Physical Characteristics
Colour Sudden Viscosity
Change
Feel
Initial Day
BF1 White No Change Smooth
BF2 White Change Smooth
After 1 Week
BF1 White No Change Tacki
After 2 Weeks
After 3 Weeks

 From the above observation, the formulation BF4 stable at room
temperature. Formulation BF1, BF2, BF3 showed changes feel.
 From the above observation , it was found that Formulation No. BF4 of
cream base obeys all the specification given in indian standard for cream.
Hence the trial no. BF4 is selected as base for the further study.

 INCORPORATION OF ACTIVE EXTRACT IN THE BASE
FORMULATION
Procedure :- Oil in water (O/W) emulsion-based cream (semisolid
formulation) was formulated. -
- The emulsifier (stearic acid) and other oil soluble components (Cetyl
alcohol, almond oil) were dissolved in the oil phase (Part A) and heated to
75° C.
- The preservatives and other water soluble components (Methyl paraban,
Propyl paraban, Triethanolamine, Propylene glycol, Herbal extract) were
dissolved in the aqueous phase (Part B) and heated to 75° C.
- After heating, the aqueous phase was added in portions to the oil phase with
continuous stirring until cooling of emulsifier took place.

Sr.No Ingredients AF1 AF2 AF3 AF4 AF5
Phase A:- Oil Phase
1 Stearic acid 15 15 15 15 15
2 Cetyl alcohol 4 4 4 4 4
4 Almond oil 4 4 4 4 4
Phase B:- Aqueous phase
3 Triethanolamine 1.0 1.0 1.0 1.0 1.0
5 Potassium Hydroxide 0.50 0.50 0.50 0.50 0.50
6 Glycerine 6 6 6 6 6
7 Methyl paraban 0.2 0.2 0.2 0.2 0.2
8 Propyl paraben 0.2 0.2 0.2 0.2 0.2
9 Water qs. 100 Qs.upto
100 ml
Qs.upto
100 ml
Qs.upto
100 ml
Qs.upto
100 ml
Qs.upto
100 ml
Herbal Extract
10 Camelia sinesis Extract 0.75 o.53 0.35 0.35 0.30
11 Punica extract 0.75 0.53 0.35 0.35 0.30
12 Aloe extract 0.75 0.53 0.40 0.35 0.35
13 Citrus aurintum 0.20 0.25 0.40 0.40 0.35

EVALUATION OF ACTIVE BASE FORMULATION
 pH of the Cream
 Viscosity
 Acid value
 Saponification value
 Irritancy test
 Accelerated stability testing
 Spreadability test
 Microbial growth test
 Total Antioxidants Capacity

 EVALUATION OF ACTIVE BASE FORMULATION
 pH of the Cream
The pH meter was calibrated using standard buffer solution. About
0.5 g of the cream was weighed and dissolved in 50.0 ml of distilled water
and its pH was measured.
 Viscosity
A 100 gm of each formulation was weighed and transpired to beaker
and the viscosity of formulation was determined with the help of Brookfield
Viscometer (DV II+ Pro model) using spindle number S64 at a 20 rpm at a
temperature of 25o C. The determinations were carried out in triplicate and
the average of three readings was recorded. Viscosity of formulation was
determine using the formula-
Viscosity (cp) = Digital reading* Factor

 Acid value
Take 10 gm of substance dissolved in accurately weighed, in 50 ml
mixture of equal volume of alcohol and solvent ether, the flask was
connected to reflux condenser and slowly heated, until sample was
dissolved completely, to this 1 ml of phenolphthalein added and titrated with
0.1N NaOH, until faintly pink color appears after shaking for 30 seconds.
Acid value = n*5.61/w
n = the number of ml of NaOH required.
w = the weight of substance.
 Saponification value
Introduce about 2 gm of substance refluxed with 25 ml of 0.5 N
alcoholic KOH for 30 minutes, to this 1 ml of phenolphthalein added and
titrated immediately, with 0.5 N HCL. Saponification value = (b-a)*28.05/w
The volume in ml of titrant = a The volume in ml of titrate = b The weight
of substance in gm = w

 Irritancy test
Mark an area (1sq.cm) on the left hand dorsal surface. The cream was
applied to the specified area and time was noted. Irritancy, erythema,
edema, was checked if any for regular intervals up to 24 hrs and reported.
 Accelerated stability testing
Accelerated stability testing of prepared formulation was conducted at
room temperature, studied for 7 days. And then the formulation studied at
45ºC ± 1ºC for 20 days. The formulations was kept both at room and
elevated temperature and observed on 0th, 5th, 10th, 15th and 20th day for
the all Evaluation parameters.

Sample was applied between two glass slides and was compressed to
uniform thickness by placing 100gm weight for 5minutes. Weight was
added to the pan. The time required to separate the two slides, i.e. the time
in which the upper glass slide moved over the lower slide was taken as
measure of spreadability. It was Calculated using the formula:
Spreadability =m*l/t
m = Weight tide to upper slide
l = length moved on the glass slide
t = time taken.
 Microbial growth test
The formulated cream was inoculated on the plates of Muller Hinton
agar media by streak plate method and a control was prepared by omitting
the cream. The plates were placed in to the incubator and are incubated at
37ºC for 24 hours. After the incubation period, plates were taken out and
check the microbial growth by comparing it with the control.

 Total Antioxidants Capacity
Total antioxidant activity was estimated by phosphomolybdenum assay
Preparation of Molybdate Reagent Solution
1 ml each of 0.6 M sulfuric acid, 28 mM sodium phosphate and 4 mM
ammonium molybdate were added in 20 ml of distilled water and made up
volume to 50 ml by adding distilled water.
Procedure:-
Hydroalcoholic extract of sample formulation in different
concentration ranging from 100 µl to 500 µl were added to each test tube
individually containing 3 ml of distilled water and 1 ml of Molybdate
reagent solution. These tubes were kept incubated at 95 ∘C for 90 min. After
incubation, these tubes were normalized to room temperature for 20-30 min
and the absorbance of the reaction mixture was measured at 695 nm.
Ascorbic acid was used as positive reference standard.

 RESULT AND DISCUSSION
 Determination of pH:
Sr No Days
Formulation
AF1 AF2 AF3 AF4 AF5
1 Initial
Day
6.2 6.3 6.4 6.4 6.5
2 7 days 6.2 6.4 6.5 6.5 6.4
3 15 days 6.3 6.4 6.5 6.4 6.5
4 21 days 6.3 6.3 6.3 6.4 6.4
5 30 days 6.5 6.5 6.5 6.5 6.6
Discussion:- The pH test was performed for base formulation for 30
days. The pH of the cream was found to be in range of 5.6 to 6.8 which is
good for skin pH. All the formulations of cream were shown pH nearer to
skin required ,But pH of formulation F1, F2,F3,F5 shows variation in pH
when stored for long period of time. Formulation F4 shows stable pH.

 Determination of Viscosity:
Sr No days
Formulation
AF1 AF2 AF3 AF4 AF5
1 Initial day 21530 cps 22500 cps 23550 cps 25590 cps 25534 cps
2 7 days 23520 cps 25580 cps 25580 cps 27050 cps 27059 cps
0
5000
10000
15000
20000
25000
30000
35000
AF1 AF2 AF3 AF4 AF5
Initial day
7 days
15 days
21 days
30 days

Discussion:-Viscosity test were performed for Active Base Formulation for 30
Days. From the observation formulation AF1, AF2 and AF3 had slightly
high viscosity, it due to concentration of active, AF4 and AF5 had
appropriate viscosity like cream.
 Acid Value and saponification value
AF1 AF2 AF3 AF4 AF5
Acid value 6.1 5.7 5.3 6.2 5.4
Saponification
value
27.1 27.3 26.3 25.8 26.8
Discussion :- From the above observation, the formulation AF4 AF3 and AF4 shows
desired acid value and saponification value.

 Irritancy test
Formulation Irritant effect Erythema Edema
AF1 NIL NIL NIL
AF2 NIL NIL NIL
AF3 NIL NIL NIL
AF4 NIL NIL NIL
AF5 NIL NIL NIL
Discussion:- All formulation shows no redness, edema, inflammation and
irritation during irritancy studies. These formulations are safe to use for skin.

 Accelerated stability testing
Sr. No. Evaluation
parameters
AF4 AF4
Room temperature Accelerated
conditions
(45oC)
1 Appearance Good Good
2 Color Light brown Light brown
4 pH 6.4 6.3
5 Consistency Semisolid and soft Semisolid and
soft
6 Viscosity 25590cps 25595cps
7 Spreadability Excellent Excellent
8 Washability Excellent Excellent
9 Irritancy test No irritation was
observed
No irritation was
observed

Discussion:- From the above observation, the formulation AF4 stable at room
temperature and accelerated condition.
Parameters
Formulation
BF1 BF2 BF3 BF4
Spreadability 26.33 ± 0.3 24.47 ± 0.4 22.35 ± 0.5 21.83 ± 0.6
 Microbial growth test
Discussion:- There were no signs of microbial growth after incubation period of
24 hours at 37ºC and it was comparable with the control.
Discussion:- From the observation, the formulation BF3 and BF4 shows
desirable spreadability.

 Total Antioxidants Capacity
PM Assay Absorbance at
695nm
Volume(l) AF4 AF3
100 0.362±0.02 0.143±0.01
200 0.753±0.05 0.333±0.04
300 1.124±0.03 0.529±0.02
400 1.451±0.03 0.871±0.02
500 1.764±0.07 1.135±0.01
Discussion- From the above observation, the active formulation 4 shows
good antioxidant capacity in the formulation

DISCUSSION
 Extract of aloe (Aloe vera) and Citrus aurintum(orange peel) Green tea
(Camelia sinensis) and pomegranate (Punica granatum)are well known for
its medicinal value in Indian traditional system of medicine and in
ayurvedic preparation.
 The herbal face cream is O/W type emulsion, hence can be easily washed
with plane water, that gives better customer compliance.
 There is a growing demand for herbal cosmetics in the world market and
they are invaluable gift of nature. Therefore, we tried to make an polyherbal
face cream containing the extract of extract of aloe (Aloe vera) and Citrus
aurintum(orange peel) Green tea (Camelia sinensis) and pomegranate
Punica granatum) in different concentration. Our study indicate that the
formulation AF4 found to be more stable while remaining formulations
were not stable and resulted in breakdown of the emulsion when stored for
longtime.
 The formulation F4 had almost constant pH, homogenous, emollient, non
greasy and easily removed after the application.

 The stable formulations were safe in respect to skin irritation and allergic
sensitization. The extracts of Green tea (Camelia sinensis) and
pomegranate (Punica granatum) provides a wealth of wonderful antioxidant
and free radical neutralizing ingredients, for example, ellagic acid, gallic
acid, punicalins and punicalagins.
 These antioxidants contain vitamin E which is required to maintain healthy
skin as well as healing and soothing to irritated skin.
 It has reported that aloesin are present in Aloe vera extract as bioactive
compound. Furthermore Aloe vera has been reported to have a protective
effect against damage to skin from ultra violet radiation due to its
antioxidant activity.
 Aloe vera contains mucopolysaccharides help in binding moisture into the
skin.
 Aloe stimulates fibroblast which produces the collagen and elastin fibers
making the skin more elastic and less wrinkled.

 CONCLUSION
- From above discussion it is concluded that the prepared formulation
showed good spreadability, no evidence of phase separation and good
consistency during the study period. From the above study it can be
concluded that it is possible to develop creams with herbal extracts.
- The ethanolic extract of Punica granatum, and Camelia sinensis exhibited
strong antioxidant activity. The results of different tests of cream showed
that the formation could be used topically in order to protect skin against
damage and prevent cream form ageing.
- Aloe’s benefits can be attributed at least partly to its nutrients, since it
contains proteins, carbohydrates (including mucopolysaccharides), vitamins
(including B1, B2, B3, B6, C, and folic acid) and minerals. These nutrients,
although beneficial individually, may work synergistically to soothe, heal,
moisturize and regenerate the skin.

Market products
A) Saundarya anti aging cream B) swati khadi Herbal anti aging cream

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