RESEARCH ARTICLE

Herbal Shampoo of Zizyphus oenoplia (L) MILL from Ethanol

Extract of Leaves
Rajesh Shukla1*, Vikas Pandey1, Dilip Golhani1, Ajay Shukla2, Kaviraj Rai3
Abstract: Artificial preservatives and detergents from time to time have been the cause of side effects. A more radical
approach in dropping the synthetic ingredients is by incorporating herbal extract whose functionality is good in comparable
with other artificial ingredients. Selection of Zizyphus oenoplia (L) MILL leaves extract was based on its particular properties
such as detergent and antimicrobial actions. This work was designed to formulate a self preserving shampoo having little
concentration of the detergent using Zizyphus oenoplia (L) MILL leaves with emphasis on safety and efficacy of chemical
ingredients and avoid the risk. Formulated and prepared the three different herbal shampoos containing dissimilar amounts
of Zizyphus oenoplia (L) MILL extract, surfactant used was sodium laureth sulfate. Physicochemical, organoleptic evaluation
and tests were performed and compared with herbal marketed product of Himalaya company. The results showed clear
shampoos were formulated in F2 and F3 products and their average pH values were in the range of 5.59-6.25, which were
suitable to hold on to the acidic cover of scalp. Results showed surface tension reduction, stable foam, wetting effect and
good cleaning and pseudo plastic rheological actions. Besides the aesthetic attributes, for example clarity and lather were
comparable with the marketed shampoos. The foam volume was on a par and formulas showed higher detergency and
foaming effects then the commercial herbal one (p 0.05). Moreover formulation can be considered safe for children due to
absence of high chemical contents.

formulate and in-vitro evaluation for detergency, foam

volume and stability, surface tension, wetting properties,
pH, viscosity, conditioning effects and stability of herbal
shampoo from Zizyphus oenoplia (L) MILL leaves and
comparison to commercial brand (Himalaya Company)
with all parameters.

INTRODUCTION
The hair shampoo is widely used amongst the other hair
care products used in daily life. Shampoos are used mainly
for cleansing the hair and scalp. The leaves Zizyphus
oenoplia (L) MILL plant, locally called as Rhamnus oenoplia
L or jackal jujube belonging to family Rhamnacae, this plant
distributed throughout the tropical parts of India, Ceylontropical Asia and Australia. [1] The roots of Zizyphus
oenoplia herbs have various activities like antiulcer,
antioxidant, [1-2] anthelminthic. [3-4] Chemical investigation
of this plant has shown the presence of cyclopeptide
alkaloids such as Zizyphus oenoplia A, B, C, D, E, abyssinine
B and A in stem bark of the plant angiogenic potential, [5]
antidenaturation and antibacteial activity, [6] wound
healing activity [7] hepatoprotective potential against
antitubercular drugs induced hepatotoxicity [8] and the part
of stomach pain medicine (pills) with the Munda tribe. [9]
Leaves numerous, 2.5-6.5 by 2-2.5 cm, ovate, acute to
mentose tips, galabarous, densely silky with hair, petioles
6-8 mm long, flower 12-20 sub sessile calyx hairy outside,
petals obviate. Fruits are edible. The bark, fruits, leaves and
stem of plant is extensively used in the rural area for
stomachache, hypotensive, low diuretic activity, wound
healing, antibacterial, anti-fungal, anti-inflammatory,
analgesic etc. Hence the ideology of preparation of a self
preserved with low concentration of a detergent will be
reducing the risk of chemicals. Sodium lauryl sulfate based
detergents are the most common but the concentration will
vary substantial from brand to brand and even within a
manufacturers product range. Objective of work was to

MATERIALS AND METHODS

Plant Collection
Leaves of Zizyphus oenoplia (L) MILL Christi were obtained
from rural area of Ranjhi, Jabalpur. Authentication and a
herbarium was submitted to Department of Botany,
Jawaharlal Krishi Vishwavidyalaya Jabalpur, M.P. Animals
(albino rats) were collected from animal house of Guru
Ramdas Khalsa Institute of Science and Technology
(Pharmacy) Jabalpur, M.P. to perform the experiment. The
Registration No. of Animal-Ethical Approval is
1471/PO/a/11/CPCSEA, India.
Chemicals and Reagents
During preparation and evaluation of the herbal shampoo
the chemicals, oleic acid (CDH, New Delhi), paraffin wax
(Merk, Mumbai), cholesterol, sodium lauryl sulfate and
formalin (Himedia, Mumbai) coconut oil (Parachute
Coimbatore) and olive oil (Allin Exporters, Mumbai) were
purchased from local market.
Preparation of Plant Extract
The collected leaves were cleaned and removed unwanted
foreign materials and accurately weighed and dried well.
Then samples were grinded and extracted with ethanol
(70% v/v). Filtered the extracts and concentrated to
dryness under reduced pressure with rotator evaporators
(40-55C) to obtain solvent-free semisolid extracts
material. Washed, weighed and packed it into artificial
containers and stored in normal temperature for the
further studies. The percentage of yield was 16.40%.

1Department

of Pharmaceutical Science, Suresh Gyan Vihar University,

Mahal, Jagatpura, Jaipur-302001, Rajasthan, India.
E-mail: rajeshshukla2628@gmail.com
* Corresponding author

2Department

of Pharmaceutical Science, Mohan Lal Sukhadiya University,

Udaipur-313001, Rajasthan, India.

3Guru

Ramdas Khalsa Institute of Science and Technology (Pharmacy),

Jabalpur-482001, Madhya Pradesh, India.

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Preparation of Herbal Shampoo

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RESEARCH ARTICLE
Table 1: Composition of the Prepared Zizyphus oenoplia (L) MILL Shampoo Formulations
Ingredients (% w/w)
Ziziphus extract
Sodium lauryl sulfate
Glycerin
EDTA
Distilled water (ml) q.s.

F0
15.00
1
0.15
100.00

F1
5
10.00
1
0.15
100.00

F2
10.00
5.00
1
0.15
100.00

F3
20
0.00
1
0.15
100.00

Table 2: Physical Appearance of the Prepared Zizyphus oenoplia (L) MILL Shampoo Formulation
Formulations
F0
F1
F2
F3
Reference

Physical Appearance
Transparent
Dark green
Dark green
Light green
Light green

Foam Producing Ability

Mild foaming
Good foaming
Good foaming
Good foaming
Good foaming

Clear four types of shampoo formulations designated as F0, F-1, F-2 and F-3 were prepared as shown in Table 1. F1,
F2 and F3 were prepared by assimilation of 5, 10 and 20%
w/w extract of Zizyphus oenoplia (L) MILL in 10%, 5% and
0% w/w of sodium lauryl sulfate. The volume was making
up to 100 ml with distilled water. As control sample, F-0
was taken (without plant extract).

5. Wetting Time
The canvas was cut into 1-inch diameter discs which having
an average weight of 0.44 g. The disc was floated on the
surface of prepared herbal shampoo solution 1% w/v and
started the stopwatch. The time necessary for the disc to
begin to sink was measured accurately and noted as the
wetting time.

Evaluation of Herbal Shampoos

To evaluate the prepared herbal shampoo formulations, as
per all the evaluation parameters like as organoleptic and
physicochemical characterization pH, solid contents and
viscosity were performed. As well to ensure the quality of
the herbal shampoo products, with surface tension, foam
volume and eye irritation, detergency, foam stability, skin
sensitization tests and preliminary stability study were
performed. The effect was compared with commonly used
marketed herbal shampoo (Himalaya Company) that was
considered as reference.

6. Rheological Property
The viscosity of the organized herbal formulations was
measured at room temperature using a digital Rheometer
(Brookfield DV-III ultra, Brookfield Engineering Lab. Inc.,
USA) fitted the spindle No. S 17 at dissimilar spindle
speeds, the best approach is optimized for measurements.
[11] Three times performed all measurements, at room
temperature and the viscosity profile of the herbal
shampoo was measured.
7. Dirt Dispersion
In a large test tube, two drops of herbal shampoo was
added which have 10 ml of distilled water and add one
drop of Indian ink. Shack the test tube for ten times. The
amount of ink in the foam was estimated like as none, light,
moderate, or heavy etc.

1. Physical Appearance/Visual Inspection

All formulated samples were observed for their physical
appearance/visual inspection. The prepared herbal
shampoo formulations were evaluated in terms of clarity,
foam producing capability and fluidity etc. [9]

8. Foam Volume and Stability

The foam volume test was performed by mixing the
prepared herbal formulas with distilled water, standard
soil and hard water (Ca++ 150 ppm). The foam was formed
by mixing in a kitchen blender. For detergency estimation
(0.25 ml of sebum 10% in hexane for 4 gm of the shampoo)
synthetic sebum was used. 40 milliliters of herbal shampoo
solution 10% w/v in distilled water was blended for 5 sec
in a kitchen blender. The height of the foam was measured
without delay and after 3 min; the test was repeated three
times in the presence of artificial sebum with standard soil
and hard water. On the other hand Ross miles foam column
method was used to evaluate the foaming stability of the
prepared herbal formulations. Briefly, 0.25% and 0.5% of
aqueous solution of each formulation were prepared and in
burette of 100 ml capacity at the room temperature. After
that independently pour from a height of 50 cm into a
measures ring cylinder and the height of produced foam

2. Determination of pH
Determination of pH of herbal shampoo formulation (10%
w/v) was done in distilled water at room temperature [10]
using digital pH meter (Mettler Teledo USA).
3. Percentage of Solid Contents
4 grams of the prepared herbal shampoo was placed in a
well clean dry evaporating dish. The liquid portion of the
shampoo was evaporated by placing on a hot plate
instrument, after that weight of the shampoo and total
solid contents after complete drying was measured.
4. Surface Tension Measurement
The surface tension measurement of the formulated
herbal shampoo (10% w/v in distilled water) was
passed out at 20C by using Nuoy tensiometer (Lauda
Germany).

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RESEARCH ARTICLE
Table 3: In-vitro Evaluation of the Prepared Zizyphus oenoplia (L) MILL Shampoo Formulations
Formula

pH

F0
F1
F2
F3
Reference

5.510.07
5.530.03
5.810.04
7.630.02
6.910.01

Solid Contents
(%)
19.110.02
22.510.02
25.110.02
28.110.02
29.110.02

Surface Tension
(dynes cm-1)
34.90.32
37.70.02
32.80.12
34.60.62
33.10.42

Wetting Time
(Sec)
2270.22
1710.19
1760.74
1590.79
1850.81

Detergency (%)

Cleaning (%)

61.100.32
76.710.12
67.670.49
63.570.26
80.120.90

18.200.02
24.500.08
32.200.05
33.610.09
34.200.02

Results are given as mean S.D. (n=5)

was measured. The foam height after 10 to 20 minutes was

also estimated. [12-13]

Reaction like as irritation test (eye), swelling of the eyelid,

ulceration, inflammation of the iris, hemorrhaging
(bleeding) and blindness etc. [14]

9. Detergency Evaluation
Thompson method [23] was used to evaluate the detergency
of the prepared herbal shampoo with minor adjustment
given that our results were evaluated gravimetrically. Hair
tresses of Asian (Indian) origin were obtained from the
market. The tresses prewashed with 5% sodium lauryl
sulfate solution and dried and cut into 10 inch, 3 gm
swatches. The sebum composition was selected to include a
variety of functional groups similar to that in actual
sebum.
The genuine formula for the artificial sebum was used in
our study, olive oil 2%, coconut oil 15% and oleic acid 15%,
paraffin wax 15% and cholesterol 20%. The hair swatch (3
gm) was floating in 20 ml of a 10% sebum solution in
hexane for 15 min with intermittent shaking, the swatch
was removed. For the determination of sebum load the
solvent was evaporated at room temperature and the hair
swatch weighted. Fifteen swatches were treated as same
and the soil levels were found in range from 9.97 to 11.06
%. Each swatch was then split into two equal samples of 1.6
gm each; one for the herbal shampoo action and the other
to act as internal control to conquer the tress-to-tress
variation in soil levels. Left untreated for control swatch.
The test swatch was washed with 0.1 ml of herbal shampoo
solution 10% using the finger method. [14] Dry the herbal
shampoo solution 10% using a hair dryer and further dried
in an oven at 60 degrees for 4 hours to ensure uniform
moisture content.
The remaining solution in the test swatch after herbal
shampoo and extracted the unwashed control swatch by
using 20 ml hexane in a stopper flask for 30 minutes on a
rotary shaker. Evaporate the hexane solution to dryness
and the sebum extracted from the test and control
swatches were weighed. According to the following
equation percentage of detergency was evaluated as of
sebum removed after shampooing:

11. Skin Sensitization Test

Animals (wistar rat) were divided into six groups (n=3).
Prior the experiment hairs of the rat were removed from
the back region. Animals of group I was served as negative
control (without any treatment). Animals group II, III and
IV were applied with herbal shampoo formulation F1, F2,
F3, respectively. While animals group V was considered as
a reference (applied with a marketed product of Himalaya
Company). Herbal shampoo was applied onto nude skin of
animal groups. A 0.8% v/v of formalin aqueous solution
was applied as a standard irritant on rats. Graded the
application sites according to a visual scoring scale, always
by the same investigate. The erythematic scale was as
follows: 0 - none, 1 - slight, 2 - well defined, 3 - moderate
and 4 - scar formation severe.
12. Preliminary Stability Study [15-17]
Preliminary Stability and acceptability of organoleptic
properties (odor and color) of herbal shampoo
formulations during storage indicate physical and chemical
stability of the prepared herbal shampoo formulations. This
study was conducted only of prepared herbal formulation
(F2). Samples were placed in dark glass test tubes and
stored in closed chamber at 45C and 75% relative
humidity. Their appearance and physical stability was
inspected and conducted for a period of 3 months at
interval of one month. To determine their mechanical
stability, every sample was centrifuged at 2400 rpm for 3
minutes and followed by its structural stability was check.
Statistical Analysis
The results were given as meanS.D. (n=5). One-way
investigation of variance (ANOVA) comparison test was
used to compare characteristics of different herbal
formulations with the commercial product Himalaya
Company. p value of 0.05 was considered to be significant.

Detergency = 100-(T 100/C)

RESULTS AND DISCUSSION

Evaluation of Herbal Shampoos
1. Physical Appearance
The herbal formulations had good characteristics with
respect to emergence of foaming. The pH of herbal
shampoo formulation has been shown to be important for
enhancing the qualities of hair, reducing irritation to the
eyes and preserve the scalp biological balance. The pH of

Where, T is the test swatch sebum weight and C is the

weight of sebum in control swatch.
10. Eye Irritation Test
Herbal shampoo solution 1% was dripped into the eyes of
six albino rats with their eyes held open with clips at the
lids. The progressive injure to the rats eyes was record at
specific interval over an average period of five minutes.

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RESEARCH ARTICLE
Table 4: Foam Volume of Shampoo Formulations

Formulas
F0
F1
F2
F3
Reference

DW
0 Min
168
153
158
175
166

3 Min
109
116
131
138
135

Foam Volume (ml)

Hard Water
0 Min
3 Min
158
105
153
101
144
123
162
114
153
114

DW + Soil
0 Min
63
57
136
92
142

3 Min
59
53
90
90
98

Table 5: Stability Study of Zizyphus oenoplia (L) MILL Shampoo (F2)

Parameters
Physical appearance/visual inspection
pH
Solids contents (%)
Surface tension (dyne. cm-1)
Viscosity (eps) at 20 rpm
Detergency ability (%)
Foaming volume (ml)

1 Month
Clear, dark green
5.510.02
22.510.02
33.220.12
67.001.89
65.120.12
160

3 Month
Clear, dark green
5.610.82
26.511.02
35.200.72
66.195.89
54.110.52
158

skeins, anchors and weight. Hence, we used the canvas disc

method for our work. Different types of canvas were used
for the test and one that gave an effective balance
between time saving and testing efficiency was selected
for the test.
The wetting ability of a substance is a function of its
concentration. By comparing the results in Table 3 with
those of detergency % in the next column it seems fair to
conclude that shampoo F3 contains the maximum
detergency and minimum wetting time and shampoos F1,
F2 and reference showed intermediate detergency and
wetting times.

herbal shampoo solution 10% in distilled water was

determined at 25C and the results are shown in Table 3.
In modern trend of formulation of herbal shampoo with
low pH, lower pH of herbal shampoo reduces damage to the
hair. Mild acidity prevents inflammation and promotes
strength of the scales, hence inducing seen. As presented in
Table 3, only shampoo F3 formulations shows a slightly
higher pH nearly the same as the marketable one, while all
the other shampoos are acid balanced and were range pH
5.6 to 5.7.
2. Solids Contents
If the herbal shampoo has too many solids it will be hard to
work on the hair or too hard to wash out. The solid
contents of the prepared herbal formulas were found 2229%. Thus they can be used, easy to wash out shampoos
(Table 3). No significant difference (p 0.05) among the
prepared formulas and the commercial brand products of
Himalaya Company (reference) was found.

5. Detergency Evaluation
The principal aim of evaluation of shampoo is removing
and cleaning of soil/sebum, experimental detergency has
been difficult to standardize, since there is no genuine
agreement on a standard soil, a reproducible soiling
procedure or the quantity of soil a shampoo should ideally
remove.
By means of greasy wool yarn by Barnette and Powers,
detergency evaluation was done in prior study. For
evaluation of detergency using 10 mg of hair clippings
obtained from salons. [18-20] Thompson et al., [14] used hair
tresses soiled with artificial sebum for evaluating
detergency. [21]
Conversely our formulas were evaluated gravimetrically
and results indicated a significant difference (p < 0.05) in
the quantity of sebum removed by different shampoos.
Shampoo F0, being a frequent-use cleanser, was expected
to have the smallest amount detergency. Shampoos F2 and
F3 also show moderate detergency and may be regarded as
mild cleansers. However shampoo F1 and reference are
active cleanser and are more appropriate for oily hair then
normal hair (Table 3).

3. Surface Tension
All prepared herbal shampoo formulations observed with
similar reduction in surface tension (p less than 0.05), as
presented in Table 3. Surface tension reduction is one of
the mechanisms of detergency. The decrease in surface
tension of water from 72.8 dynes cm-1 to 32-37 dynes cm-1
by the shampoos reflects of their good properties of
detergent action.
4. Wetting Action
The rate of wetting or the wetting ability of surface-active
agents is normally used to find out their comparative
efficacies. Even though the Draves test [19] has been
adopted as the official test, the canvas disc wetting test [20]
offers a number of advantages over it. The canvas disc
technique is more accurate and time reducing then the
official test. For the canvas test also, the different materials
and apparatus required which is easily available in the
laboratory, unlike the Dreves test, [19] which requires cotton

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2 Month
Clear, dark green
5.531.02
24.110.92
32.520.32
65.280.99
67.100.10
160

6. Cleaning Action
Cleaning action of herbal shampoo was evaluated on wool
yarn in grease. Though cleaning or soil/sebum removed is

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RESEARCH ARTICLE
the main aim of a shampoo, experimental detergency
evaluation has been difficult to standardize, as there is no
genuine concord on a standard soil, a reproducible soiling
process or the amount of soil a shampoo should preferably
remove. As given in the results, there is a major difference
in the amount of sebum removed by the different shampoo.
The final herbal formulation results of detergency showed
that has significantly similar detergency ability, when
compared to the marketed preparations and it was found in
between 19-33 % (Table 3).

The all herbal shampoo formulation showed no skin

sensitization including marketed shampoo except F1
formulation. Other additives are the second most common
cause of skin reactions besides fragrances. In the majority
cases, these were only soft or transient such as stinging
sensation. In few cases, reaction may be more severe with
redness, edema.
12. Preliminary Stability Study
Study for the suitability of organoleptic properties (odor
and color) of the selected F2 during the storeroom period
indicated complete chemical and physical stability of the
prepared herbal shampoo formulation. The stability of the
formulated herbal shampoo formulation is shown in Table
5.

7. Dirt Dispersion
Herbal Shampoo that causes link to concentrate in the foam
is considered as poor quality product, the dirt should not
stay in the foam because it will be difficult to rinse away. All
three formulated herbal shampoos (F1, F2 and F3) showed
similar results as compared the marketed one, except the
control (F0). These results illustrate that no dirt would
stays in the foam, so prepared formulations were
satisfactory. [22-23]

CONCLUSION
The current study intended to prepare a stable, self
preserved herbal shampoo formulation which contains less
concentration of detergent to reduce the risk of skin
infection. No significant difference between both F2 and F3
in comparison with the reference marketed shampoo was
seen regarding the physicochemical properties and in-vivo
evaluation.

8. Viscosity
Product of viscosity plays a vital role in physical properties
like as shelf life stability and product aesthetics such as
clarity ease of flow on removal from packing a spreading on
application to hair and product consistency in the package.
The flow properties of non-newtonian formulations are
usually not measured with a single data point, since their
viscosity is reliant on the shear rate. The best way is to take
multipoint measurements determined to those of the
process being modeled. [24-25]
Shampoo F0 and F1 indicates low viscosity products,
whose viscosity remains almost unchanged over the range
of r.p.m. used. Shampoo F2 and F3 illustrate pseudoplastic
behavior same as the commercial one (reference), which is
a pleasing quality in a shampoo preparation. On raising the
shear, the viscosity drops, which would allow ease of
distribution on the hair.

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9. Foam Volume and Stability

All the prepared herbal shampoo showed similar foaming
characteristics in distilled water as well as hard water. Only
shampoo F2 and F3 foamed well as the commercial one in
the presence of soil (Table 4). In a practical point of view,
shampoos F0 and F1 would not foam well with first hair
wash, once the sebum has been removed all five shampoos
would show comparable foaming properties. A point to be
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detergency and foaming with cleaning property, which only
confirms the fact that a shampoo that foams well need not
clean well.
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There was eye irritation test on application of herbal
preparation. The animal recovered well in 3 days same as
the case with the reference. No eyes irritation, dryness or
scaling caused by the tested formulations.
11. Skin Sensitization Test

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Acknowledgment
Authors are thankful to Guru Ramdas Khalsa Institute of
Science and Technology, Jabalpur M.P. to provide the
laboratory for performing the tests and computer labs for
technical support.
Cite this article as: Rajesh Shukla, Vikas Pandey, Dilip
Golhani et al. Herbal Shampoo of Zizyphus oenoplia (L)
MILL from Ethanol Extract of Leaves. Inventi Impact:
Cosmeceuticals, 2016(1):50-55, 2016.

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