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DOI: 10.2174/1874372202115010052, 2021, 15, 52-58
The Open Dermatology Journal
Content list available at: https://opendermatologyjournal.com
Development and Evaluation of Herbal Hair Serum: A traditional way to
Improve Hair Quality
Ruchi Tiwari1,* , Gaurav Tiwari1, Ajeet Yadav2 and Vadivelan Ramachandran3
1Department of Pharmacy, Pranveer Singh Institute of Technology, Kanpur, India
2Department of Pharmaceutical Sciences, J.S. University, Shikohabad, Firozabad-283135, India
3Department of Pharmacology, JSS College of Pharmacy, JSS Academy of Higher Education & Research, Ooty, Nilgiris, Tamil Nadu, India
In the mammalian system, the hair follicle is known to be the most significant organ that determines appearance, gender distinction, provides
intense temperature protection, and plays a role in self-defense. The younger generations have begun to suffer from extreme hair loss problems due
to many lifestyle-related changes such as fatigue, anxiety, intake of junk foods, use of different hairstyling/coloring methods, etc. The loss of hair
is not temporary in most cases, but it results in alopecia. Many people suffering from hair loss are in search of multiple treatments due to extreme
anxiety and tension, from mythology to traditional and therapeutic healing to the use of minoxidil and finasteride. To improve hair growth and to
prevent hair loss, hair root activation is required.
The present study was intended to use different herbs to formulate herbal hair serum for general purposes (hair application).
Crude herbs are obtained from nearby regions. In order to obtain the extract, the required part of herbs, such as Citrus sinensis peel, Zingiber
officinale roots, and Linum usitatissimum seeds, Nigella sativa, and Trigonella foenum-graecum were specifically weighed and dispensed in water,
boiled, allowed to cool, and then filtered. To the filtrate, castor oil and vitamin E were added. The formulated herbal hair serum was tested, and
different criteria were determined and recorded in this text, such as physical appearance, viscosity, pH, homogeneity, eye sensitivity (Draize eye
test), hair growth activity, hair weight, stability test, etc.
Prepared herbal hair serum was found to be pale brown in color with pseudoplastic behavior. The texture of the formulation was smooth and
lubricative, and the pH was within the acceptable limit. Herbal hair serum revealed good hair growth as well as hair weight after application with
minor sensitivity for the first time. The formulation was found to be stable for seven days.
Herbal cosmetics are still commonly used by average citizens because of fewer side effects and greater protection and safety profile. Current
research has revealed that herbal formulations are effective in enhancing hair consistency.
Keywords: Citrus Sinensis, flaxseed, Nigella sativa, Zingiber officinale, castor oil, Trigonella foenum-graceum.
Article History Received: March 1, 2021 Revised: April 2, 2021 Accepted: April 30, 2021
Hair growth is based on a dynamic and precisely regulated
mechanism that is not fully understood. It is a cyclical mecha-
nism involving hair shaft synthesis, elongation, and ultimately
* Address correspondence to this author at the Department of Pharmacy, Pranveer
Singh Institute of Technology, Kanpur, India; E-mail: email@example.com
shedding. Human hair usually comprises follicles of anagen,
catagen, and telogen phases. The hair follicle in the anagen
phase actively accumulates cytochrome and easily develops the
hair shaft. When the hair follicle matures into the anagen hair
follicle, the telogen hair follicle is not able to develop neonatal
hair shafts . Alopecia Areata (AA) is a recurrent immune-
mediated skin disorder with a non-scarring loss of hair. In the
Development and Evaluation of Herbal Hair Serum The Open Dermatology Journal, 2021, Volume 15 53
general population, it has an incidence range of 1.7 percent and
2.1 percent, with higher prevalence in young patients (21 40
years) and no significant difference in incidence between males
and females reported. This condition may have a detrimental
effect on the quality of life of patients in a manner close to that
reported for other skin diseases such as dermatitis and
psoriasis. AA's pathogenesis can be explained by the
inflammatory cascades involving a breakdown of the hair
follicle's immune privilege, associated with T lymphocyte
invasion, and an autoimmune process that creates autoantigens
from proteins associated with melanogenesis. In addition,
different variables, including family backgrounds, genetic
bases, environmental factors, illnesses, medications, injuries,
potential emotional stress, and oxidative stress, can be
implicated in AA pathogenesis . Defective antioxidant
defense or exaggerated expression of reactive oxygen species
(ROS) has been reported to predispose to oxidative stress,
which is believed to play a significant role in many
inflammatory skin disorders such as AA. In this respect,
several studies have documented that AA has a detrimental
effect on the blood and skin tissue's oxidant-antioxidant
balance. AA's latest clinical treatments are centered primarily
on immune regulation using numerous forms of medications,
including topical and systemic steroids, phototherapy, and
immune suppressants such as cyclosporine and methotrexate;
meanwhile, certain vitamins and micronutrients are expected to
play a role in the treatment of AA and can alleviate oxidative
Citrus sinensis (orange peel) has shown remarkably
positive results on hair. Daily use can make the hair smoother
and shinier with these peels. They are filled with antioxidants
and help to treat the harm caused by pollutants as well. Daily
use of orange peels, filled with a lot of vitamin C, makes the
hair bouncy and shiny and reduces dryness and dullness of
hair. In addition, orange peels, a huge source of vitamin B12
and vitamin E, assist in hair development. They also delay the
pace at which the hair begins to turn gray [4, 5].
The richest plant source of the ω-3 fatty acid is flaxseed,
i.e., “α- linolenic acid” (ALA). Flaxseed oil is low (9%) in
saturated fatty acids, high (18%) in monosaturated fatty acids,
and rich in polyunsaturated fatty acids (73%) . The possible
health effects of flaxseed oil, fibers, and flax lignans include
minimizing coronary disease, atherosclerosis, asthma, cancer,
obesity, osteoporosis, autoimmune and neurological disorders.
In addition, flaxseed is full of fatty acids and antioxidants that
tend to remove pollutants and dead cells from the scalp.
Flaxseed gel can be added as a moisturizer to the scalp and
hair, which can help promote growth and boost the quality of
the current hair. Flaxseed gel is super hydrating, has
conditioning benefits, and, without the crunch, makes the hair
very fluffy [7, 8].
Nigella sativa (NS) is an annual flowering plant native to
the Mediterranean countries, Pakistan, and India, commonly
known as black cumin. The key pharmacologically active
components of NS are thymoquinone (TQ), dithymoquinone
(DTQ), thymohydroquinone (THQ), and thymol (THY) .
Inhibition of NF-kB activation has been reported to inhibit
inflammatory cell infiltration in the brain by TQ, which
constitutes the predominant portion (30% - 48%) of NS. By
inhibiting cyclooxygenase 2 (COX-2) expression and
prostaglandin D2 (PGD2) synthesis, TQ demonstrates anti-
inflammatory behavior, which will explain why NS has been
considered a natural treatment for many illnesses and
inflammatory disorders . In addition, recent studies have
shown that PGD2 has a significant inhibitory function in the
hair follicle. In particular, its binding to the GPR44 receptor
expressed in the hair follicle seems to cause follicle
miniaturization. This contributes to the growth and
preservation of vellus-like hair and is likely to inhibit stem
cells from maturing into progenitor cells and vellus-like hair
from maturing into natural, dense, and pigmented, terminal
hair. Consequently, NS inhibition of NF-kB and PDG2
activation and synthesis can contribute to the regulation of the
hair cycle. Nigella sativa, a flower native to Eastern Europe,
Southwest Asia, and the Middle East, contains black seeds.
Black caraway, black cumin, black onion seeds, and kalonji are
widely recognized. These seeds have been used in treatments
for allergy, asthma, diabetes, headache, weight loss, arthritis,
and intestinal worms for thousands of years. Nigella sativa seed
is an ideal ingredient for medicine and cosmetics, according to
a 2016 study. The research identifies the properties of black
seed oil as antibacterial, antifungal, anti-inflammatory, and
antioxidant. Advocates of black seed oil for hair claim that by
treating concerns such as dandruff while maintaining the scalp
moisturized, these properties can help in protecting the scalp.
Since it is anti-inflammatory, when diluted with a carrier oil to
soothe scalp conditions that cause inflammation, flakiness, and
discomfort, the black seed oil is perfect . It also destroys
fungal infections and viruses, so it can help people with lice.
Black seed oil is an adjuvant, like lavender, lemon, and
rosemary essential oils, ensuring it combines well with other
ingredients to exhibit medicinal effects. Black seed oil is anti-
rheumatic, and it is very helpful in alleviating joint injuries that
arise with age and athleticism. Furthermore, black seed oil can
help to regrow hair in thinning regions on the scalp. Nigella
sativa oil has commonly been used for thousands of years for
the treatment of hair loss .
Fenugreek seed extract contains a dietary supplement that
also contains micronutrients such as B-vitamins, antioxidants,
and trace elements that are also present in the hair. Fenugreek
(Trigonella foenum-graceum L.) is a leguminous herb.
Fenugreek is native to Southern Europe, Western Asia, and the
Mediterranean region. Fenugreek seeds contain a wide range of
active ingredients such as saponins, especially diosgenin,
yamogenin, and gitogenin derivatives, trigonelline alkaloids,
flavonoids, galactomannan vitamins, and fiber, etc . The
seeds have a good fragrance and a sour flavor. As both a
culinary and medicinal plant of the ancient world, it has a long
tradition. Because of their therapeutic ability, including their
usage as hypoglycemic, antiulcerogenic, hypo-cholesterolemic,
and antihypertensive agents, the beneficial properties of
fenugreek seeds have drawn significant interest in the recent
past . There are known beneficial effects of fenugreek on
hair growth, but the mechanism is not characterized. It is
addressed that fenugreek interacts in a physiological manner
with the synthesis of DHT (dihydrotestosterone) by enhancing
the blood supply of hair follicles and steroid saponins. The
54 The Open Dermatology Journal, 2021, Volume 15 Tiwari et al.
impact of DHT on genetically predisposed hair follicles is
suspected to be one cause for male and female variant hair loss.
Binding DHT to the hair follicle results in the hair's progressive
miniaturization and subsequent hair loss [15, 16].
Ginger, one of the most commonly used spices worldwide,
is the root of Zingiber officinale (L.) Rosc. It contains several
active ingredients, including gingerol, zingerone, shogaol, and
beta bisabolene. It has been used in various diseases such as
rheumatoid arthritis, neurodegenerative diseases, inflammation,
and asthma as folk medicine. In addition to high antioxidant
activity and reduced expression of certain pro-inflammatory
biomarkers, many of its active constituents have anti-diabetes,
anti-cancer, and anti-inflammatory functions. In addition,
ginger is helpful in the treatment of dandruff and irritated, itchy
scalp. It has natural anti-inflammatory and antiseptic properties
that tend to keep the skin clean and healthy. Overall, ginger
stimulates the growth of hair, prevents hair from thinning, and
renders hair to be glossy and smooth [17, 18].
Castor oil, also known as Ricinus communis, is a vegetable
oil that is rich in nutrients and comes from castor beans. While
it originates from tropical East Africa, it is commonly used in
India and the West Indies for cosmetic and medicinal purposes.
Castor oil, which is known as black castor oil, is typically
transparent or pale yellow unless it is derived from roasting or
boiling. Castor oil is a long-chained fatty acid at the molecular
level. “Fatty acids give hair follicles essential proteins and
nutrients and prevent inflammation of hair follicles,” as
according to Bridgette Hill. Castor oil has a few other
noteworthy effects in addition to being anti-inflammatory.
“Increased gloss or shine of the hair, increased hair shaft
flexibility by coating the hair shaft, and antimicrobial
properties are some of the benefits of using castor oil.” A total
of 90% of castor oil is made up of ricinoleic acid fatty acid. It
is believed to activate the Prostaglandin E2 receptor, which can
contribute to the dilation of the blood vessels. Increased blood
vessel dilation increases the supply of oxygen-rich and
nutrient-rich blood to the follicle's powerhouse, dermal papilla,
or hair root .
The present study intended to use extracts of different
herbs to formulate herbal hair serum for the improvement of
2. MATERIALS AND METHODS
2.1. Collection of Plant Part
For the preparation of herbal hair oil, various plant
materials were collected viz., Citrus sinensis, Linum
usitatissimum, Nigella sativa, Zingiber officinale, and
Trigonella foenum-graecum from the Medicinal Plant Garden
of Pranveer Singh Institute of Technology, Kanpur, Uttar
Pradesh, India, and were properly authenticated in the
Department of Pharmacognosy.
2.2. Formulation of Herbal Hair Serum
Table 1 presents the different ingredients used in the
formulation of herbal hair serum. All fresh herbs, such as
Citrus sinensis peel, Zingiber officinale roots, Linum
usitatissimum seeds, and Nigella sativa, Trigonella foenum-
graecum were specifically weighed and dispensed in 500 ml of
water. The contents referred to above were boiled for 15 min.
They were permitted to cool after 15 minutes of boiling and
then filtered. Castor oil and vitamin E were added to the
filtrate. Afterward, the Prepared serum was kept in a spray
2.3. Evaluation of The Hair Serum
2.3.1. Physical Appearance
The physical appearance, color, and feel of the prepared
herbal hair serum are visually tested. Table 2 reflects the
2.4. Homogeneity Test
A clean and dry object glass was smeared with the hair
serum, and a cover glass was sealed. The appearance under the
light of some coarse particle/homogeneity was investigated.
Herbal hair serum was tested by visual examination for
homogeneity and tested for some lumps, flocculates, or
2.5. pH Test
The pH meter was calibrated using pH 4 and pH 7 buffer
solutions. Then, the electrode was soaked in the hair serum and
left until the pH normalized after a few minutes .
The viscosity measurement was performed with spindle
number 6 on a Brookfield viscometer (RVDV-II+PRO). In the
beaker, 50 ml of hair serum was placed, and the viscosity was
measured at various rpm, i.e., 10, 20, 50, 100 .
Spreadability was measured by a parallel plate process
typically used to assess and measure the spreadability of semi-
solid preparations. One gram hair serum was pressed between
two horizontal plates of dimension 20× 20 cm, the upper of
which weighed 125 g. The spread diameter was measured after
1 min. Spreadability was calculated using the following
S = M × L / T
Where, S= Spreadability, M= Weight in the pan (tied to the
upper slide), L= Length moved by the glass slide, and T =
Time (in sec) taken to separate the slides completely .
Table 1. Ingredients used in the preparation of herbal hair
Ingredients Plant part Quantity (%)
Peel of fruit
Development and Evaluation of Herbal Hair Serum The Open Dermatology Journal, 2021, Volume 15 55
Table 2. Evaluation parameters of herbal hair serum.
Physical appearance Yellowish-brown
Viscosity (cps) * 10 RPM 20 RPM 50 RPM 100 RPM
6800±0.011 3950±0.023 1760±0.009 1110±0.052
Draize eye test Mild irritation
Hair length (mm)*
(Herbal hair serum)
7th Day 14th Day 21st Day
Weight of the hair (mg) on 3rd week of the treatment 224.98
*the value is expressed as mean ±SD, n=3.
Table 3. Stability study on herbal hair serum.
pH* Viscosity at 100 rpm (centipoise) *
Initial Final Initial Final
6.7±0.022 6.6±0.012 1110±0.002 1124±0.117
*the value is expressed as mean ±SD, n=3.
2.8. Test the Sensitivity of the Eye (Draize Eye Test)
Eye sensitivity was tested on three rabbits for getting the
average result of sensitivity. One drop of hair serum was
injected into the left eyes of the rabbits in physiological NaCl
(as the control is the right eye) and then observed for 30
minutes, 60 minutes, 120 minutes, 240 minutes, 1 day, 2 days,
3 days, and 4 days. The scores of the cornea, iris, and
conjunctiva were determined .
2.9. Hair Growth Activity Test
This test was conducted on 3 areas of a single rabbit on
either foot, right and left of the shaved backs of the rabbits,
each 4x4 cm in size. Then, after 3-5 minutes, depilatory cream
(Veet ® cream) was applied, and the area was rinsed with
water until the area was clean with hair. As an antiseptic, 70
percent ethanol was then added. Before any action evaluation,
the rabbit was left behind for 24 hours. As no intervention was
performed, treatment 1 was the standard control, and treatment
2 was the positive control where the hair serum with the test
material was added. Again, as no intervention was undertaken,
treatment 1 was the standard control, and treatment 2 was the
positive control where hair serum with the test material was
added. For 3 weeks, 0.1 ml of each drug was then administered
twice a day to the rabbit. The first day of use of the hair serum
was called day 0 [5, 6].
2.10. Qualitative Assessment of Hair Growth
Qualitative measurement of hair growth analysis was
carried out by visual inspection of 2 criteria, initial hair growth
time (minimum time needed for hair growth on the shaved
area, measured from the darkening of the skin color showing
initial hair growth) and completion time for hair growth
(minimum time required for the entire shaved area to be
covered with new hair)  on three groups (Group I: Normal
control; Group II: Negative control; Group III: Positive control
(Herbal hair serum)). Each group consists of three rabbits.
2.11. Observations of the Growth of Hair
It was done by taking 10 random hair strands on each box
on days 7, 14, and 21. The hair had been drawn out,
straightened, and stuck with the tape, and measured using
Mitutoyo Digimatic wireless calipers. The average length
collected was analyzed to see if the difference between the
research region and the control was statistically significant [4,
2.12. Hair Weight Measurements
To evaluate the weight of each box, the hair was removed
and weighed on day 21 and then statistically measured [3, 20].
2.13. Microbial Contamination
Microbial contamination of hair serum was determined via
spreading a thin loopful of the serum on a nutrient and
Sabouraud agars and incubating for 48 hours at 37°C. In order
to examine the degree of contamination, one gram of material
was used to be dispersed in 4 ml of sterile Ringer solution,
containing 0.25% Tween 80. In the same dispersion vehicle,
sufficient dilutions were made, and 0.5 ml was mounted on the
appropriate solid medium using the viable surface procedure.
Following required incubation, emergent colonies were
counted [23, 24].
The herbal hair serum was kept for three months at two
separate temperatures of 4± 2°C and 30± 2°C, with 65% RH.
Compared with the original pH and viscosity, the pH and
viscosity of the herbal hair serum were determined after three
3. RESULTS AND DISCUSSION
3.1. Physical Appearance
It was observed that the color of all the herbal hair serum
was pale brown with a translucent look, which on the
application was found to be smooth.
56 The Open Dermatology Journal, 2021, Volume 15 Tiwari et al.
By visual examination of the appearance and presence of
any lumps, flocculates, or aggregates, the produced herbal hair
serum was checked for homogeneity. The homogeneity of
prepared serum has been shown to be fine.
3.3. Rheological Study
The rheological examination of prepared herbal hair serum
was conducted, and it was observed that viscosity was reduced
as the shear rate increased (Fig. 1). The pseudoplastic flow was
also described, which is a beneficial attribute for topical herbal
hair serum since it guarantees optimum area coverage upon
application. The mechanistic explanation for the flow
behaviour found is as follows: the long-chain molecules of the
polymers are spontaneously arranged in dispersion under
standard storage conditions. These molecules tend to organize
their long axes in the direction of the force exerted when
applying shear stress. This stress-induced orientation reduces
the internal resistance of the material. In addition, the solvent
molecules earlier associated with the polymer molecules will
also be released. Thus, the effective concentration and size of
the molecules are low. The material allows a progressive
increase in the shearing stress .
3.4. pH Determination
The pH of the whole herbal hair serum was 7.3, which was
sufficient for the hair, suggesting that the herbal hair serum
was suitable for the hair .
3.5. Draize Eye Test
To evaluate the protection of the herbal hair serum, a
sensitivity assessment test was performed. To determine the
ocular sensitivity index, the test was conducted on 3 rabbits.
Tests on the left eye were conducted with controls on the right
eye. In physiological saline, 3 drops of sterile 2.5 percent
herbal extract were added to the left eye of the rabbit, and
measurements were taken at 30 minutes, 60 minutes, 120
minutes, 240 minutes, 1 day, 2 days, 3 days, 4 days. There was
no image of opacity in the left eye, normal conjunctiva, no
swollen eyelid, and the iris image appeared normal. It was seen
that the eyes created tears. The index of ocular annoyance was
2 on the first day and 0 on the second, third, and fourth days
(maximum ocular irritation index of 110). Thus, it can be
inferred that the herbal serum possesses a moderate
inflammatory effect .
Fig. (1). 7KHUKHRORJLFDOH[DPLQDWLRQRISUHSDUHGKHUEDOKDLUVHUXP.
Development and Evaluation of Herbal Hair Serum The Open Dermatology Journal, 2021, Volume 15 57
3.6. Hair growth activity test
Table 2 reveals that normal control showed a very close
outcome with negative control for the first week, while positive
control showed little difference. However, in areas treated with
positive regulation, there is a substantial hair length difference
between both normal control and negative control hair.
However, a major difference was reported by comparing the
growth behavior displayed by normal control, negative control,
and positive control. The discrepancy between the groups was
found to be important (p<0.05). Thus, it was found that,
although using a single formula, the hair serum used in this
study displayed a more important hair growth activity of 6.98
mm. In Table 2, it can be seen that in the third week, herbal
hair serum induced a greater weight of the hair [21, 22].
3.7. Microbial Contamination
At the end of seven days, the microbial contamination of
the herbal hair serum after 24 hours was observed to be 1.89
CFU for fungi .
3.8. Stability Studies
Table 3 reveals that the herbal hair serum was stable during
the research time, as these serums showed no physical
instability, and there was no noticeable difference in the pH
before and after the study [23, 24].
Results have shown that herbal hair serum provides various
essential nutrients needed to preserve the proper function of the
sebaceous glands and support the growth of natural hair. In the
personal hygiene and health care system, the use of herbal
cosmetics has changed by several folds. Therefore, the herbal
cosmeceutical individual care or personal health care industry,
which is actually concentrating and paying extra care on the
production of herbal-based cosmetics, has a considerable
clamor. As nowadays, in the coming years, it is a fast-
developing market with a mammoth scope. In cosmetic
formulations, the use of bioactive ingredients has a valuable
impact on body characteristics and offers nutrients that are
important for preserving good and beautiful hair. It can be
inferred that prepared herbal hair serum has a beneficial effect
on the mechanism of hair growth and increased consistency.
Medicinal plants have been used for the treatment of hair
diseases since antiquity because of fewer side effects and
hypersensitivity reactions. It is time to dump the chemical-
laden hair care products in favour of natural alternatives. The
traditional system of medicine in India acclaims a number of
herbal drugs for hair growth promotion. The best part is that
herbal extracts will provide microprotein supplements to hair
and provide enough nourishment, resulting in safe and sound
hair. Herbal cosmetics have become increasingly common in
the personal care industry, and there is a high demand for them
in everyday life due to their lack of parabens and sulphates.
The global herbal industry is projected to be worth more than
US$10 billion dollars, and it is increasing at a rate of three to
four percent per year due to increased demand. In terms of
production and demand of herbal products, Europe is the
largest region, followed by Asia.
ETHICS APPROVAL AND CONSENT TO
HUMAN AND ANIMAL RIGHTS
No animals/humans were used in the studies that are basis
of this research.
CONSENT FOR PUBLICATION
AVAILABILITY OF DATA AND MATERIALS:
The data that support the findings of this study are
available from the corresponding author [R.T] upon a
CONFLICT OF INTEREST
The authors declare no conflict of interest, financial or
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© 2021 Tiwari et al.
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