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Shata – Dhauta – Ghrita – A Case Study

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  • Aimil Pharmaceuticals

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Shata-dhauta-ghrita (SDG) is an Ayurvedic preparation, commonly prescribed for treatment of wounds, burns, chicken pox, scars, herpes, leprosy and other skin diseases. It is prepared by washing clarified butterfat (cow ghee) one hundred times with water. Having potential as a topical product for the treatment of skin diseases, it needs evaluation in modern scientific parameters. The study was carried out to evaluate the changes taking place while washing cow ghee one hundred times which makes it an important topical product.
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Indian Journal of Traditional Knowledge
Vol. 8 (3), July 2009, pp. 387-391
Shata- dhauta- ghrita – A case study
Supriya Deshpande1*, Avinash Deshpande2, Suresh Tupkari & Anoop Agnihotri2
1Delhi Institute of Pharmaceutical Sciences and Research (DIPSAR), New Delhi, 110 017; 2Dr DY Patil Pratishthan’s, Padmashree Dr
DY Patil Institute of Pharmaceutical Sciences and Research, Pimpri, Pune 411 018, Maharashtra
E-mail: supriyadeshpande149@yahoo.co.in
Received 13 march 2008; revised 15 September 2008
Shata–dhauta–ghrita (SDG) is an Ayurvedic preparation, commonly prescribed for treatment of wounds, burns, chicken
pox, scars, herpes, leprosy and other skin diseases. It is prepared by washing clarified butterfat (cow ghee) one hundred
times with water. Having potential as a topical product for the treatment of skin diseases, it needs evaluation in modern
scientific parameters. The study was carried out to evaluate the changes taking place while washing cow ghee one hundred
times which makes it an important topical product.
Keywords: Cow ghee, Topical preparation, Skin hydration, Ointment base, GC/MS spectrum
IPC Int. Cl.8: A61K36/00, A61P17/00, A61P17/02, A61P17/08
Traditional system of medicine, evolved over the
ages, had been completely looking after the healthcare
of the world until the advent of allopathic system of
medicine. As this system used knowledge of modern
biology and chemistry, for both discovery and
treatment, it found fast acceptability and now
occupies significant space in the area of healthcare. In
spite of this, the contribution of the traditional
systems of medicine to healthcare continues to be
enormous. The acceptability of traditional
preparations, which are normally polyherbal, is
increasing because of the general impression that
these products are benign. The single molecule based
drugs used in allopathic system can have severe
adverse effects1. Panchagavya, i.e. 5 products
obtained from cow (cow milk, curd, ghee, urine and
dung) mentioned in Ayurveda have significant role in
disease management2. Cow ghee is having various
activities by itself and also by formulations developed
using cow ghee. For example, effect of Bramhi ghrita
on CNS, sedative and anticonvulsant activity of
Unmadnashak ghrita, wound healing activity of
Darvhi ghrita, wound healing activity of Hingvadya
ghrita, antiinflammatory activity of Jatyadi ghrita,
etc3-8. Shata-dhauta-ghrita (SDG) is 100 times
washed (shata = one hundred, dhauta = washed)
clarified butterfat. It is mentioned in traditional texts
for the management of conditions like burns, chicken
pox, scars, wounds, herpes, leprosy, and other skin
diseases and as a vehicle for drugs to be applied
externally9,10. In the study, the Ayurvedic preparation
has been evaluated for its physicochemical parameters
and changes occurring during washing were analyzed.
An attempt is made to find out the rationale behind
washing cow ghee 100 times with water.
Methodology
All the chemicals were purchased from E Merck,
SD Fine Chemicals, Mumbai, Universal Laboratories
and Qualigens Fine Chemicals. All the solvents were
distilled before use. Solvents used for UV, IR and
GC/MS studies were of spectroscopy grade. Cow
ghee was procured from Go Seva Anusandhan
Kendra, Kusumba, Jalgaon. The copper vessels were
purchased from local market. GC/MS was carried out
on Shimadzu QP–2010 at Shimadzu Service Centre,
Toshvin Analytical Laboratory, Mumbai. All the
results are obtained by repetition of the each
experiment at least 3 times. Cow ghee was observed
for its organoleptic properties and analyzed for
various physicochemical parameters (moisture
content, viscosity, acid value, saponification value,
iodine value, unsaponifiable matter, Rechert Meissl
value, Polenske value, etc.) prescribed for lipids in
Pharmacopoeias (IP, USP). Particle size and copper
content were also determined11–18. For GC/MS
analysis, Fatty Acid Methyl Esters (FAME) of the
samples was prepared. About 25 ml of 10% solution
______________
*Corresponding author
INDIAN J TRADITIONAL KNOWLEDGE,VOL 8, No.3, JULY 2009
388
of sample was prepared in n– hexane. In a suitable
flask, 19 ml of this solution was treated with 1.25 ml
of 3M methanolic potassium hydroxide and the flask
was shaken for 30 min, the mixture was placed in a
large centrifuge tube containing 10 ml of saturated
solution of sodium chloride. The flask was rinsed
several times with distilled water in to centrifuge tube
(Use of water keeps eventual methanolic peaks in
chromatograph to a minimum). Shaking for 30 min
and then centrifuging brought the hexane solution
containing the esters to the top of the tube. This
hexane solution was transferred to a labeled, stoppered
bottle. A GC/MS separation of the esters was
performed19. Conditions for GC/MS included
Model - Shimadzu QP 2010, Column - 30 m/0.32
mm id/0.25 micron BP 20, Column temperature -
50°C to 260°C at 5°C per minute, Injector
temperature - 250°C, Split ratio – 40, Ion source
temperature - 220°C, MS interference entioned in
Bhaishajya-kalpana-vidnayan9,10 .
The copper vessels were cleaned thoroughly and
rinsed with distilled water. 2.5 kg of previously
standardized cow ghee was taken in copper vessel.
1.5 L of distilled water was added to it. 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 1.5 L distilled water was
added in previously washed cow ghee and similar
procedure was repeated. This operation was carried
out one hundred times to obtain SDG. Samples
were collected after each washing and stored in
plastic containers at room temperature for analysis.
Analysis of SDG as per Pharmacopoeial standards
for oils and fats was carried out based upon
physicochemical parameters. Being a fatty product,
the tests applied for fats and oils were carried out to
know the physical and chemical properties of
finished product (SDG). Tests carried out for
standardization of raw material (organoleptic
properties, moisture content, viscosity, acid value,
saponification value, iodine value, unsaponifiable
matter, Rechert Meissl value, Polenske value
particle size and copper content, etc.) were
performed for evaluation of SDG11-18. GC/MS
analysis was carried out to find out the changes
occurred in fatty acid composition. FAME were
prepared and subjected for GC/MS analysis using
method similar to that used for raw material.
Results and discussion
A comparison of physicochemical parameters of
cow ghee and SDG has been done (Table 1, Fig. 1).
GC/MS spectra for FAME of cow ghee and SDG
were obtained (Figs. 2&3) and comparison of the area
per cent of major peaks was done (Table 2, Fig. 4).
From the study carried out, some facts behind
Table 1— Comparison of physicochemical parameters of cow ghee and SDG
Parameter Cow ghee SDG Change
Organoleptic properties
Colour Golden yellow White
Odour Characteristic, Pleasant Odorless
Taste Characteristic Tasteless
Texture Granular, oily Smooth, non - oily
Homogeneous mass
Physical parameters
Moisture. content (%) 0.27 0.8 2.98 times
Ph 4.6 5.9 1.28 times
Particle size (µ) 179.02 59.99 2.98 times
Viscosity (cp) at 20 rpm for 30 sec 8000 9770 1.22 times
Copper content (ppm) 0.13 1.2 8.86 times
Chemical parameters
Acid value 0.84 0.11 7.59 times
Saponification value 234.26 25.96 9.02 times
Iodine value 35.59 2.82 12.62 times
Unsaponifiable matter 0.78 0.04 18.72 times
RM value 21.67 0.22 98.50 times
P value 1.1 0.1 11 times
Note : increase, : Decrease
DESHPANDE et al.: EVALUATION OF AYURVEDIC DRUG SHATA- DHAUTA- GHRITA
389
preparation of SDG can be illuminated. Initially, the
pure lipid phase, i. e. cow ghee comes in contact with
aqueous phase. Due to trituration, it results in
formation of w/o type of emulsion as lipid phase (cow
ghee) is a major phase. As the washing continues, due
to pressure applied during trituration, particle size of
fat granules gets reduced (as evident from decrease in
particle size). Eventually with successive washings,
aqueous phase becomes predominant over lipid phase
(evident from increase in moisture content).
This results in the phenomenon of phase inversion
resulting in o/w type of emulsion. Chances are there
that it may result in complex system like w/o/w
emulsion. The characteristic odor and granular, oily
consistency in cow ghee remains no longer in SDG
making it homogeneous, smooth, non-oily product,
which is easier to apply, thus improves patient
compliance.
Increase in moisture content may be useful for skin
hydration and cooling effect, which can justify its use
for the treatment of burns20. pH change from acidic to
neutral makes it beneficial to prevent skin irritation.
Fig. 1— Physicochemical parameters of cow ghee and SDG
INDIAN J TRADITIONAL KNOWLEDGE,VOL 8, No.3, JULY 2009
390
So, the preparation can be applied on open wounds.
Reduction in particle size makes the product non-
granular, non-sticky, homogeneous, which makes it
easy to apply on skin and may result in increased rate
of absorption through skin. Viscosity was found to be
increased. Washing results in homogeneous mass of
oil in water emulsion with better consistency and
viscosity which is helpful in its topical application.
Increase in copper content of the preparation makes it
significant in removal of scars and as an
antiinflammatory agent10,21. Decrease in iodine value
indicates decrease in degree of unsaturation, which
eventually reduces chances of rancidity thus increase
stability of the product16,17. Where as decrease in RM
and P value indicate decrease in steam volatile water
soluble and water insoluble fatty acids, respectively.
These mainly include short chain fatty acids18.
Decrease in unsaponifiable matter is the indicator of
removal of matter other than fatty acids15.
GC/MS spectrum of cow ghee (Fig. 2) showed its
composition which lies in the standard values
prescribed2. In GC/MS spectrum of SDG (Fig. 3),
percentage of almost all fatty acids was found to be
decreased. Decrease in degree of unsaturation can be
supported by absence of unsaturated fatty acids, viz.
oleic acid and linolenic acid in SDG. Preparation of
SDG can be correlated with the phenomenon of fat
splitting. Fat splitting is the process in which, fat is
hydrolyzed in the presence of water to yield free fatty
acids and glycerols (Fig. 5)22. The reaction proceeds
step by step and is reversible. If reactants and
products are not removed from the sphere of the
reaction, an equilibrium depending upon the
concentrations of the former will eventually be
reached. In practical fat splitting, a high degree of
hydrolysis is ensured by using a large excess of water
and repeatedly withdrawing the glycerol rich aqueous
phase and replacing it with fresh water. High
Fig. 2— GC/MS spectrum of FAME of cow ghee
Fig. 3— GC/MS spectrum of FAME of SDG
Table 2— Fatty acid composition of cow ghee and SDG
(Area % of GC/MS peaks)
Fatty acid Cow ghee SDG
Butyric 0.65 38
Caproic 1.13 0.66
Caprylic 0.77 0.41
Capric 1.68 0.84
Lauric 2.03 0.16
Myrestic 7.17 0.31
Palmetic 18.74 0.24
Stearic 11.8 0.18
Oleic 14.3 0
Linoleic 1.7 0.63
Linolenic 0.43 0
Fig. 4— Fatty acid composition of cow ghee and SDG
Fig. 5— Fat splitting process
DESHPANDE et al.: EVALUATION OF AYURVEDIC DRUG SHATA- DHAUTA- GHRITA
391
temperature and pressure accelerates aqueous
hydrolysis. In preparing SDG, temperature is not
increased but there is repeated and prolonged tritutation
of the fat and water mixture. Thus, the pressure factor
may be contributing for fat splitting in this case. After
each trituration, aqueous phase is withdrawn and
replaced by fresh slot of the same in case of fat
splitting. It has also been found that splitting is
accelerated by presence of mineral acids, certain metal
oxides and sulphonic acids. SDG has been mentioned
to be prepared in copper vessels. Thus, incorporation of
copper metal in the mixture during trituration was
indicated by increase in copper content in SDG.
Copper can act as a catalyst to promote fat splitting.
The above hypothesis of fat splitting by using large
excess of aqueous phase can be correlated with the idea
of washing cow ghee 100 times with water. It can be
pointed out that by washing cow ghee 100 times with
water, the triglycerides are splitted into glycerol and
free fatty acids. The decrease in acid value can be
correlated with the fact of fat splitting in to glycerol
and fatty acids, which are removed along with the
aqueous phase. From the present work, it can be
concluded that changes taking place in cow ghee while
washing it with water 100 times to prepare Shata-
dhauta-ghrita, makes it an elegant and suitable product
for topical application. However, there is a need to
carry out further study on its stability, pharmacological
evaluation as an effective topical product and its
formulation development.
Acknowledgement
Authors owe a deep sense of gratitude to
Mr R Girijan, Toshvin Analytical Laboratory,
Shimadzu Service Center, Mumbai, for making
GC/MS facility available and also helping in
interpretation of data.
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... In our earlier studies, we have evidenced the effect of 'Murcchana' process to ensure maximum acceptability, stability and better shelf life of ghrita preparation [11]. 'Shata-Dhauta' is a process involves washing of purified ghee one hundred times with water which increases stability of ghrita and makes it elegant and suitable product for topical application [12]. ...
... In present research work, attempt was made to prepare cow ghee based Polyherbal Ghrita formulation as per Ayurvedic texts. Based on therapeutic (wound healing) potential, present herbs of Western Ghats, India were selected to prepare herbal extracts (Kalka) [12,13]. The name 'Bhallatakadi' was assigned from the synonymous name of S. anacardium. ...
... The possible decomposition of fatty acids during storage can lead to bad odour and rancidity which affects stability and shelf life of ghrita therefore prepared polyherbal ghrita formulation was processed with ancient Ayurvedic procedures viz. 'Murcchana' and 'Shata-Dhauta' samskara [12,36,37]. ...
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Modern formulation science requires all excipients to be inert and to conform to Pharmacopoeial standards. However, Ayurveda, the ancient science of health from India believes that all substances possess therapeutic actions. Modern formulators have a choice of choosing from the modern formulation bases (e.g.; vanishing cream base) or Ayurvedic bases (e.g.; cow ghee) while formulating an herbal product. Nonetheless, all such bases must conform to specifications of physico-chemical properties that have the potential to affect the efficacy/safety/stability of the product. In this study, we have compared the physical characteristics of a modern base (vanishing cream) with two Ayurvedic bases (clarified butter fat -cow ghee and Shata-dhauta-ghrita). Shata-dhautaghrita (SDG) is prepared by washing cow ghee 100 times with water and is prescribed for treatment of wounds, burns etc. All three bases were evaluated for their pH, viscosity, spreadability, organoleptic properties, globule size, acid value, saponification value, peroxide value, ester value, iodine value and free fatty acids. It was found that while cow ghee and vanishing cream base had comparable characteristics, SDG exhibits a much less degree of unsaturation (suggesting better physico-chemical stability) and better consistency (and hence suitability for topical applications). The paper also suggests the possible mechanism for improvement of these characteristics in the process of conversion of cow ghee to SDG. It further suggests that SDG might be a potential candidate as a base for topical preparations, esp. for wounds and burns.
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The aim of present investigation was to study the therapeutic potential of Honey, Daruhaldi and Shatdhaut ghrut and honey singly and in combination on experimental excision wound healing activity in rats. Excision wounds of about 500 mm2 and 2 mm depth were used for the study. Parameters studied were period of epithelization, rate of wound contraction and time required for 50% wound closure (CT-50). Rats treated with Honey + Daruhaldi combination showed better wound healing activity as compared to other test drugs. Effect of this combination on wound area was less as compared to povidone iodine. However effect of Honey + Daruhaldi & Povidone iodine were comparable in % wound contracture or % wound closure. In this Excision wound model shatdhaut Ghrut alone or in combination with Honey showed less wound healing activity as compared to povidone iodine. The improved wound healing potential and synergistic effect of Honey & Daruhaldi can be attributed to additive antioxidant potential of Honey and flavonoid moities in Daruhaldi combination showed better wound healing potential than Honey & Daruhaldi alone.
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Darvhi ghrita is a herbal formulation containing cow's ghee and Berberis aristata as its main constituents. It is being tested in present study for its topical wound healing activity. The activity was studied in male wistar rats. In this study two wound models were studied viz. Incision wounds (for tensile strength) and Excision wound (To quantify wound contraction with histopathological evaluation). Treatment with Darvhi ghrita alone promoted wound contraction and reduced the time for closure showing good healing potential compared to marketed Framycetin sulphate cream (FSC) 1% w/w. The Histopathological studies reveal complete healing with test formulation showing better epithelization, fibrosis and angiogenesis. The present study demonstrates wound healing potential of test formulation.
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The present study aims to evaluate the wound healing activity of a polyherbal formulation Hingvadya ghrita in rats. Incision wounds for tensile strength and excision wounds for wound contraction along with the histopathological examination of the regenerated tissue were employed to investigate the wound healing potential. Topical application of the test formulation alone promoted the tensile strength (incision wounds) and wound contraction (excision wounds) showing healing potential comparable to Framycetin sulphate cream (1% w/w). Histological examination reveal good keratinization, epithelization, fibrosis and collagenation indicative of the wound healing potential of Hingvadya ghrita. The present study thus offers a valuable insight into the claimed wound healing potential of the test formulation.
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Rev. ed. of: Vogel's textbook of quantitative inorganic analysis. 4th. ed. 1978 Incluye bibliografía e índice
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OBJECTIVE: To eavaluate the CNS activity of Bramhi Ghrita, a polyherbal formulation containing Bacopa monneri , Evolvulus alsinoids , Acorus calamus , Saussurea lappa and cow′s ghee. MATERIALS AND METHODS: The effect of Bramhi Ghrita on motor coordination, behavior, sleep, convulsions, locomotion and analgesia was evaluated in mice using standard procedures. RESULTS: The formulation exhibited reduced alertness, spontaneous locomotor activity and reactivity. It also antagonized the behavioral effects of d-amphetamine, potentiated the pentobarbitone-induced sleep and increased the pain threshold. Bramhi Ghrita protected mice from maximum electroshock and pentylene tetrazole-induced convulsions. CONCLUSION: The Bramhi Ghrita was found to be a CNS depressant with anticonvulsant. activity