INTRODUCTION:
In Ayurveda, the five cow-derived products known as
panchgavya1 i.e. Cow Milk (Dugdha), Curd (Dahi),
Clarified Butter Fat (Ghee), Urine (Go-Mutra) and Dung (Gobar)
are the essential components for the treatment of many illnesses. Fig. 01
depicts the Panchagavya elements2,3.
Fig. 01: Five Elements of Panchgavya
Ayurveda remains one of the most ancient
healthcare system and yet a live tradition practiced widely in India, Sri Lanka
and other countries that has a sound philosophical and experiential basis4. Ayurveda has several formulations for management of
aging and skin care. Ayurvedic literature describes more than 200 herbs,
minerals and animal products for skin care. For this study, Ghee of indigenous
cow, leaves of Tulsi (Ocimum sanctum) and Neem (Azadiracta indica)
and rhizome of Haridra (Curcuma longa) are used, shown in Fig. 02. Skin
care properties of all the three herbs were reported5,6,7,8. Traditionally prepared cow ghee has two types of
properties Samshodhana (cleansing or detoxifying)9 and Samshana (palliative)10. Such ghee alone or in combination with honey11 is considered to be extremely useful for treating
wounds, inflammatory swellings and blisters for promotion of quick healing.
Ghee is used in many Ayurvedic traditional preparations and also as an ointment
base.
MATERIALS AND METHODS:
Materials:
The herbal material Turmeric, Neem and Tulsi powder
were obtained from a local authentic herbal distributor of Wardha, Maharashtra.
All other chemicals were purchased from lobachemie Pvt. Ltd. and Merck life
science Pvt. Ltd.
|
|
|
|
Clarified Butter Fat (Cow Ghee)
|
Ocimum sanctum (Tulsi)
|
|
|
|
|
Azadirachta indica (Neem)
|
Curcuma longa (Haridra/Haldi)
|
Fig. 02: Ingredients used in the Formulation
Methodology:
a) Preparation of shata-dhauta-ghrita by
niragni method:
The thoroughly cleaned copper vessels were rinsed with
distilled water. Then 100 g previously standardized cow ghee and 200 mL of
distilled water was added in to the copper vessel. With the help of manual
copper agitator, the mixture of cow ghee and water were triturated for 5–8
minutes. The contents in copper vessel were allowed to settle. Water was
decanted carefully, avoiding loss of ghee. The fresh slot of 200 mL distilled
water was added in previously washed cow ghee and similar procedure was repeated for Hundred times obtain SDG 12. It
results in butter like odourless white silky cream which has the incredible
property of penetrating all the seven layers of the skin without blocking the
pores 13. Dasha-Dhauta-Ghrita
(DDG) samples were collected
after 10 washings and Shata-Dhauta-Ghrita (SDG) samples were collected
after 100 washings of ghee, then both DDG and SDG were stored in plastic
containers at room temperature for further analysis. The physical properties of
finished product SDG based panchgavya polyherbal cream were analysed as per
Pharmacopoeial standards for oils and fats. Additionally, standardization of
Ghrita, Dasha-Dhauta-Ghrita and Shata-Dhauta-Ghrita including organoleptic
properties, moisture content, viscosity, pH, particle size and copper content
were performed14,15,16. Table 01 depicts the comparative data of
organoleptic properties of Ghrita and Shata-Dhauta-Ghrita. For
the measurement of Ghee and water during washing in certain intervals was
followed as shown in Fig 03.
Fig. 03: Measurement of Dhaut Ghrita and Mardan
Process Decanted Water
b) Herbal extract preparation:
The rhizome of Turmeric (Curcuma longa)17,18,19, leaves of Tulsi (Ocimum sanctum Linn)20,21 and leaves of Neem (Azadirachta indica)22,23 were cleaned properly with water, dried and powdered
using an electric mixer, sieved and stored in an air tight container for
further use. All the powdered herbal drugs (5g each) were soaked in 100mL
distilled water separately in 3 conical flasks (Fig. 04) for 24 hours
with constant shaking for first 8 hours and then filtered filter paper (ARYAN
BRAND Grade no KP1 Hand Made Filter Paper)24,25,26.
Fig 04: Herbal Extracts of Haldi, Neem and Tulsi in
water
c) Preparation of oil phase:
6 g myristic acid was taken in a 250mL beaker and it
was kept on hot water bath at 70˚C, after complete dissolution of myristic
acid, 5g of Shata-Dhauta-Ghrita was added to it and mixed by continuous
stirring, 3g of glycerol monostearate was added with continuous stirring, and
then 3g of cetyl alcohol was added and stirred continuously to avoid lumps
formation.
Fig 05: Preparation of Oil Phase
d) Preparation of aqueous phase
23.3mL of each of the extracts (Neem, Turmeric and
Tulsi) were added to 10g of glycerine in a beaker and placed on hot water bath
at 70˚C. 2.5g triethanolamine was added to it with continuous stirring,
then 0.2g of Methyl paraben was added to it with gentle stirring.
e) Addition of aqueous phase to oil phase:
The aqueous phase was poured in to the oil phase
slowly with continuous stirring at 70 ˚C. The mixture was allowed to cool
to room temperature with continuous stirring. Later 2 mL of perfume was added
to the mixture, and then transferred the cream to container sterilized with
99.5% ethanol. The cream was stored in cool and dry place. Final products are
made in five prototypes F1, F2, F3, F4 and F5 are shown in Fig 06.
Fig 06: Prepared Shata-Dhauta-Ghrita based
creams.
Table 03: Finalized composition of SDG based Panchgavya
polyherbal cream
|
S. No.
|
Ingredients
|
Quantity
|
|
1.
|
Myristic acid
|
6 g
|
|
2.
|
Shata-Dhauta-Ghrita
|
5 g
|
|
3.
|
Glycerol
monostearate
|
3 g
|
|
4.
|
Cetyl alcohol
|
3 g
|
|
5.
|
Distilled water
based herbal extract
|
70 mL
|
|
Neem
|
23.3 mL
|
|
Turmeric
|
23.3 mL
|
|
Tulsi
|
23.3 mL
|
|
6.
|
Glycerine
|
10 mL
|
|
7.
|
Triethanolamine
|
2.5 g
|
|
8.
|
Methyl paraben
|
0.2 g
|
f) Evaluation of cow ghee based poly herbal
panchgavya cream for physical parameters27, 28, 29
· Appearance: The appearance of the Cream was found by observing its
colour, texture etc.
· pH: The pH of Shata-Dhauta-Ghrita based poly herbal
panchgavya cream was determined with the help of Auto digital pH meter at 25şC
by preparing 20% 100 mL solution of cream; the pH meter was calibrated and
measured.
· Determination of Homogeneity: The prepared formulation was tested for the
homogeneity by visual appearance and by touch.
· Spread ability test: 500 mg of the cream was sandwiched in
between two glass slides. Weight of 100 g was placed on upper slide of glass
slide. The weight was removed and extra cream was scrapped off. The lower slide
was fixed on board of apparatus and upper slide was fixed with non-flexible
string on which 20 g load was applied. Noted down the time taken by upper slide
to slip off.
S = m × 1/t
Where, S- spread ability; m- Weight tied to upper
glass slide; l- Length moved on a glass slide; t- Time taken
The Spread ability test was carried out in triplicate
and the average of the readings was recorded.
· Smear type: The test was conducted after the application of Shata-Dhauta-Ghrita
based poly herbal panchgavya cream on the skin
· Emolliency test: Emolliency of cream was checked by
application of fixed amount of cream on skin.
· Dilution test: This test can be conducted by adding water or oil to
the emulsion. If water is added, and
it gets mixed with the emulsion, then the emulsion is of the oil-in-water (o/w)
type. If water does not mix with the emulsion, then it is known as the
water-in-oil (w/o) type.
· Dye solubility test: In the dye solubility test emulsion was
mixed with the water soluble dye (amaranth) and observed under microscope. If the emulsion becomes red, it means that
it is the oil-in-water type and vice-versa.
a)
Comparative
evaluation of anti-bacterial activity of prepared shata-dhauta-ghrita based
polyherbal panchgavya cream and marketed samples30
To test the antibacterial activity of newly formulated
Shata-Dhauta-Ghrita based Polyherbal Panchgavya cream, the gram-positive
bacteria Staphylococcus aureus was selected. Before starting the test,
the bacterial strain was cultured in nutrient agar plates and incubated in
incubator for 18 to 24 hrs at 37şC temperature to obtain bacterial
colonies. After overnight incubation, colonies on the plates were selected with
autoclaved sterile inoculating loop of inoculating needle and transferred it to
a glass test tube of sterile physiological saline which was prepared for test
and mixed it thoroughly. The growth of bacteria is measured by the turbidity of
saline solution. The turbidity of bacterial suspension was compared with 0.5 Mc
Farland standard solution which produces turbidity equivalent to
1.5×108 CFU/mL (Colony forming units). Antimicrobial susceptibility of
Formulated vanishing cream was tested by using a well-diffusion method
(National Committee for Clinical Laboratory Standards). The formulations were
tested on Mueller Hinton plates to detect the presence of bacterial growth. All
plates were inoculated with the test bacteria i.e. Staphylococcus aureus
which have been previously adjusted to 0.5 Mc Farland standard solution
turbidity using pour plate method. The inoculated plates were dried for around
5 minutes to remove excess moisture if any in laminar air flow chamber by
placing the plates near to burner to avoid any contamination. 50ul aliquots of
each test and standard marketed cream with different concentrations (0.3
mg/10mL, 0.6mg/10mL and 0.9 mg/10mL) were added into each well after the
inoculation of each plate with bacteria. The well was made on agar by using
sterilized cork borer. The plates were sealed, labelled and placed in an
incubator at temperature of 37şC for 24 hours. After24h of incubation, each
plate was observed for inhibition zones. Inhibition zone was measured by using
ruler (Scale). The zones were measured in millimetres and the reading was noted
down. The results of the anti-bacterial activity of prepared formulation of Shata-Dhauta-Ghrita
based Polyherbal Panchgavya Cream and standard i.e. marketed formulations
Pond’s and Vicco against S. aureus at 3 different concentrations are
captured in Fig 09, Fig 10 and Fig 11 respectively.
b)
Comparative
evaluation of anti-fungal activity of prepared shata-dhauta-ghrita based
polyherbal panchgavya cream formulation and marketed samples31
The fungal organism Saccharomyces cerevisiae
was selected to study the anti-fungal activity of prepared vanishing cream. The
fungal organism was cultured in Potato dextrose agar medium at 27 şC room
temperature for 3-5 days. After that the colonies of S. cerevisiae were
observed after 5days and they were diluted further and compared with 0.5 Mc
Farland which were prepared earlier, which is equal to 1.5×108 CFU/mL.
The same procedure of antibacterial activity test were followed for antifungal
activity testing of newly prepared vanishing cream only the organism and agar
were changed. All plates were inoculated with the test fungi i.e. S.
cerevisiae which have been previously adjusted to the 0.5 Mc Farland
standard solution by pour plate method in PDA medium. Cream anti-fungal
susceptibility was tested by using well-diffusion method (National Committee
for Clinical Laboratory Standards). 50 µl aliquots of each test and standard
marketed cream formulations with different concentrations (0.3 mg/10mL,
0.6mg/10mL, and 0.9mg/10mL) were added into each well after the inoculation of
the plates with fungi. The well was made on agar by using sterilized cork
borer. The plates were sealed, labelled and placed in an incubator set at 22°C
for 4-5 days. After 5 days of incubation, each plate was observed for
inhibition zones. Inhibition zone was measured by using ruler (Scale). The
zones were measured in millimetres and the reading was noted down. The results
of the anti-bacterial activity of prepared formulation of Shata-Dhauta-Ghrita
based Polyherbal Panchgavya Cream and standard i.e. marketed formulations Pond’s
and Vicco against S. cerevisiae at 3 different concentrations are
captured in Fig 09, Fig 10 and Fig 11 respectively.
RESULTS:
Ocimum sanctum, Curcuma longa and Azadirachta indica are well
known for their medicinal and cosmeceutical value in Indian traditional system
of medicine. In the present work, it was decided to formulate Panchgavya
Polyherbal Multipurpose Cream composed with all these three herbal extract
incorporated in Shata-Dhauta-Ghrita as cream base. While making Shata-Dhauta-Ghrita,
after each washing of Cow ghee, the weight of ghee, volume and time required
for washing were recorded. (See Supplementary Material, Table 07)
a)
Evaluation of Shata-Dhauta-Ghrita
based Polyherbal Panchgavya Cream for physical parameters:
Assessment of Shata-Dhauta-Ghrita based poly
herbal panchgavya cream was done for various physical parameters as per
official guidelines. Evaluation of physical parameters of F1, F2, F3, F4 and F5
is recorded in Table 04 and comparative physical parameters study between best
batch and standard is recorded in Table 05.
Appearance:
The appearance of the Cream was found by observing its
colour, texture and found to be smooth.
pH: The
pH of Shata-Dhauta-Ghrita based poly herbal panchgavya cream was
determined with the help of Auto digital pH meter at 25şC by preparing 20% 100
mL solution of cream, the pH meter was calibrated and measured pH was found to
be 7.14 as shown in Fig 07.
Fig. 07: pH estimation of Panchgavya Polyherbal
Shata-Dhauta-Ghrita Cream
Determination of Homogeneity:
The prepared formulation was tested for the
homogeneity by visual appearance and by touch the cream was found to be
graceful.
Spreadability test: 500 mg of the cream was sandwiched in between two
glass slides. Weight of 100g was placed on upper slide of glass slide. The
weight was removed and extra cream was scrapped off. The lower slide was fixed
on board of apparatus and upper slide was fixed with non-flexible string on
which 20 g load was applied. Noted down the time taken by upper slide to slip
off. The experimental observation is shown in Fig 08.
Fig 08: Spreadability estimation of Panchgavya
Polyherbal Shata-Dhauta-Ghrita Cream
Smear type: The test was conducted after the application of Shata-Dhauta-Ghrita
based poly herbal panchgavya cream on the skin and smear formed was
non-greasy.
Emolliency test: Emolliency of cream was checked by application of
fixed amount of cream on skin and found that no residue left after application.
Dilution test: In the dilution test cream was diluted with water and
the emulsion was found to be o/w type as it was diluted with water.
Dye solubility test: In the dye solubility test emulsion was mixed with the
water soluble dye (amaranth) and observed under microscope. The Scattered
globules appear red and continuous phase was colourless, so according to this
observation it is identified as w/o type emulsion.
Fig. 09: Evaluation of Antibacterial and Antifungal
activity of SDG based Panchgavya Polyherbal Cream
Fig. 10: Evaluation of Antibacterial and Antifungal
activity of Vicco marketed cream
Fig. 11: Evaluation of Antibacterial and Antifungal
activity of Ponds marketed cream
Antibacterial and Antifungal study:
The herbal formulation prepared was tested for
antimicrobial activity against selected strains and compared with standards.
Formulation shown best result which was almost equal to creams taken as
standards. Results obtained from the antimicrobial study showed that the
developed formulation have an inhibitory effect on the S. aureus
and S. cerevisiae. The in vitro antimicrobial activities were
calculated in term of the zone of inhibition diameter (mm) the result recorded
in Table 06.
Stability study:
The developed cream was subjected to the accelerated
stability study for 6 months. Results reveal that the evaluated parameters such
as colour, consistency, fungal growth, smoothness, viscosity, spreadability,
and pH did not change considerably when kept at 25 ± 2°C and at 40 ± 2°C. There
is no microbial growth or phase separation in any of the conditions. The
stability study is in progress for 2 years.
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. In results, the formulations show no redness, edema, inflammation and
irritation during irritancy studies.
DISCUSSION:
A Shata-Dhauta-Ghrita (SDG) infused Panchgavya Polyherbal Cream
was developed during this study using combination of traditional ayurvedic
techniques along with the scientifically validated ingredients. Total 5
variants F1, F2, F3, F4 and F5 were developed, amongst all of them batch F5
exhibited favourable physicochemical properties smooth texture, optimal pH,
enhanced viscosity, and non-greasy smear making it more ideal as compared to
plain ghee, SDG and other 4 formulation. Antibacterial and antifungal tests
showed that the SDG-based cream performed comparably, and in some cases better,
than marketed creams like Vicco and Ponds. This is attributed to the
synergistic action of herbal extracts (Tulsi, Neem, Haldi)
and SDG with its deep skin-penetrating, hydrating properties and its natural,
non-irritant base, improved spreadability with additional benefits from copper
content and panchgavya integration that makes our formulation superior than
previous studies and commercial products related to SDG. Unlike synthetic
bases, SDG acts as both a carrier and an active therapeutic agent, enhancing
skin health while ensuring safety and user comfort. These results validate SDG
as an effective base for multifunctional herbal creams. Further studies on
stability and clinical effectiveness are recommended to advance its use in
modern skincare.
This study sheds light on the underlying processes
involved in the preparation of SDG. The core ingredient cow ghee which is a
pure lipid phase firstly interacts with the aqueous phase and on trituration it
produces water-in-oil (w/o) emulsion. As washing proceeds the pressure is
applied during trituration that reduces particle size of fat granules and the
aqueous phase progressively overtakes the lipid phase with each washing, as
evidenced by an increase in moisture content. Phase inversion from this change
produces an oil-in-water (o/w) emulsion. Additionally, a complex w/o/w emulsion
system could develop. SDG produces a homogenous, smooth, and non-oily product
that eliminates the distinctive smell and grainy, oily texture of cow ghee.
This increased uniformity improves patient compliance and ease of use. Its use
in burn treatment is supported by the possibility that the higher moisture
content will cool the skin and help hydrate it. The product also benefits from
a pH shift from acidic to neutral, which lowers the possibility of skin
irritation and allows it to be applied to open wounds. The product is easier to
apply and may improve skin absorption because of the reduced particle size,
which also makes it non-granular, non-sticky, and consistent. Increasing
viscosity during the washing process results in a more uniform oil-in-water
emulsion with better consistency, which is beneficial for topical application.
SDG's increased copper content is noteworthy since it has both
anti-inflammatory and scar-removal properties, which make it a potent cream
basis in natural cosmetics.
CONCLUSION:
The present study successfully demonstrates the formulation and
evaluation of a Shata-Dhauta-Ghrita (SDG) infused Panchgavya Polyherbal
Cream, enriched with standardized extracts of Ocimum sanctum, Curcuma
longa, and Azadirachta indica. The integration of these three
potent Ayurvedic herbs with SDG (traditional lipid base) known for its deep
dermal absorption has resulted in a stable, non-greasy, cosmetically elegant
formulation with multifunctional therapeutic potential. Among the five
prototypes developed, Batch F5 exhibited the most favourable physicochemical
properties, including optimal pH, enhanced viscosity, superior spreadability,
and smooth, non-sticky texture. The formulation also showed significant
antibacterial and antifungal activity, comparable to or exceeding that of
marketed creams such as Vicco and Ponds. The synergistic combination of herbal
extracts amplified the individual properties of each plant, delivering enhanced
moisturizing, wound healing, anti-aging, anti-wrinkle, UV-protective, and
antimicrobial benefits. The use of SDG as a natural cream base not only
improved skin compatibility but also contributed to the overall efficacy and
aesthetic quality of the product. Comparative evaluation with commercial
formulations validated the safety, stability, and performance of our cream.
These findings highlight the potential of SDG-based polyherbal creams as
effective, natural alternatives to conventional cosmeceuticals. Further
research, including long-term stability testing, dermal absorption profiling,
and clinical trials, is recommended to establish its broader applications in
dermatology and personal care.
ACKNOWLEDGEMENTS:
The Authors are grateful to Mahatma Gandhi
Institute for Rural Industrialization, Wardha, Maharashtra and Ministry of
MSME, Government of India, New Delhi for constant support and providing
research facilities.
FUNDING:
This research study was funded by Mahatma
Gandhi Institute for Rural Industrialization (MGIRI), Wardha-442001,
Maharashtra, Ministry of MSME, Government of India.
COMPETING INTERESTS:
The authors declare that they have no known
competing financial interests or personal relationships that could have
appeared to influence the work reported in this paper.
Author Information:
Authors and Affiliations: Bio-Processing
and Herbal Division, Mahatma Gandhi Institute for Rural Industrialization,
Wardha, Maharashtra, India; Jaykishor A. Chhangani, Soumya G. Katre, Payal S.
Kathale, Shamli R. Jamane, Adarsh Kumar Agnihotri and Ashutosh A. Murkute.
Corresponding Author:
J. A. Chhangani, Bio-Processing and Herbal
Division, Mahatma Gandhi Institute for Rural Industrialization, Wardha,
Maharashtra, India.
CONTRIBUTIONS:
Conceptualization, literature review and
study design: J.A.C.; Original draft writing, schematic representation, Data
collection, referencing, citations and editing: S.G.K.; Data collection,
Practical Procedures, Analysis, figures and tables: P.S.K.; Anti-microbial
analysis: S.R.J.; Review and editing: A.K.A. and A.A.M., All authors read and
approved the final manuscript.
DATA AVAILABILITY:
The datasets used or analysed during the
current study are available from the corresponding author on reasonable
request.
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