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Indo American Journal of Pharmaceutical Research, 2017 ISSN NO: 2231-6876
PRELIMINARY QUALITATIVE SCREENING, PHYSICOCHEMICAL STANDARDIZATION AND
ANTIMICROBIAL ACTIVITY OF A HERBO MINERAL SIDDHA FORMULATION GANDHAGA
KARUPPU
D. Deepa Bai1,2, Ashima Joshi1,2, Geeta Tyagi1,2, V. Vijaya Kumar1, S. C. Verma*1, M. B. Shankar1
1Pharmacopoeia Commission for Indian Medicine and Homoeopathy, Kamala Nehru Nagar, Ghaziabad, 201002. Uttar Pradesh,
India.
2Central Council for Research in Siddha, Chennai-106.
*Corresponding author
Dr. S. C. Verma, *Dr. D. Deepa Bai
Principal Scientist
Pharmacopoeia Commission for Indian Medicine and Homoeopathy,
Kamala Nehru Nagar, Ghaziabad, 201002.
scvpharma@gmail.com
*Senior Research Fellow (Siddha)
deepasiddhaa@gmail.com
Copy right © 2017 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
ARTICLE INFO
ABSTRACT
Article history
Received 05/07/2017
Available online
02/08/2017
Keywords
Standardization,
Gandhaga Karuppu,
AYUSH,
Authenticate,
Traditional Siddha Medicine,
Antimicrobial Activity.
A recent resurgence of interest in the traditional systems like AYUSH (Ayurveda,Yoga,
Unani, Siddha & Homoeopathy) systems of Indian Medicine is due to the fact that these
systems are natural/ environmental friendly/ biodegradable. The safety based efficacy of these
medicines is also equally important because of the issues of adulteration and substitution and
contamination and for globalization too. Therefore, the development of pharmacopoeial
standards becomes mandatory as emphasized by WHO. Under the Law of drugs and cosmetic
act 1940, quality control and analytical standardization of ASU&H drugs in the light of
phytochemical aspects geared up in India. Scientist’s (silver bullet) method of extracting the
chemical to target the effect led to suggestion that most drugs are devoid of activity which
may due to the in-vitro methods and lack of solubility of the drug. This paves way to support
traditional formulations, where drug parts are added as such and processed and the finished
product administered with suitable adjuvants (Gunshot method to treat the individual as a
whole). The objective of the present study to evaluate the organoleptic characters, acid and
basic radicals, pH, phytochemicals, standardize physico chemically to authenticate the
sulphur based classical Siddha formulation Gandaka karuppu and also to evaluate the
antimicrobial activity. The Results revealed, the presence of sulphate, chloride, carbonate,
fluoride and iron and ammonium, acidic pH, no significant antimicrobial activity , higher acid
insoluble ash values which might be due to higher mineral content present in the finished
product. The Organoleptic characters and other physicochemical values signify the good
quality and purity of the herbo-mineral drug Gandhaga karuppu.
Please cite this article in press as Dr. D. Deepa Bai et al. Preliminary Qualitative Screening, Physicochemical Standardization
And Antimicrobial Activity Of A Herbo Mineral Siddha Formulation Gandhaga Karuppu. . Indo American Journal of
Pharmaceutical Research.2017:7(07).
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INTRODUCTION
The World Health Organization estimates that about 80% of the populations living in the developing countries rely almost
exclusively on traditional medicine for their primary health care needs [1]. Herbal medicines were in great demand in the developed as
well as in developing countries for primary health care because of their wide biological and medicinal activities, higher safety margin,
and lower costs [2]. The Siddha system is practiced in Tamil Nadu since antiquity. But the scientific evidence to prove the rationale
of using these formulations in health care is not well established. To widen the selection of traditional remedies globally and to make
the expectation of ‘well health’ of humanity more reasonable, all the drugs are to be standardized, pertaining to the Laws of regulation
by the AYUSH Ministry, Government of India. Since adulteration and substitution remain challenging, scientific documentation
becomes essential for the reference standards and so, the present study is to standardize the Siddha classical compound formulation
Gandaka karuppu (GK)[3] and correlate it clinically since solubility of the Sulphur remains challenging and discuss it with the anti
microbial activity.
MATERIALS AND METHODS
Formulation Composition
Gandhagam (Sulphur) and four Plant drugs mentioned in the Table below serve as ingredients of the Siddha formulation
Procurement and authentication of drugs
The raw drugs were procured from raw drug store from local market of Chennai, India. The identity and authenticity of the
drugs were confirmed by Dr. M. Allimuthu, Professor and HOD, Department of Gunapadam, Govt. Siddha Medical College,
Arumbakkam, Chennai.
‘Suddhi’ (Purification) of Crude drug ingredients of Gandaga karuppu
Table.1 Method of Purification of Crude drug ingredients of Gandaga karuppu.
S No
Crude drug ingredients
Part used
Method of Purification
1
Sulphur
The market crude drug
Melted with clarified butter in an iron pan and poured
through a clean muslin cloth into cow’s (Bos taurus
indicus) milk. The process is repeated thrice. The
obtained sulphur is then soaked in,
1. Fresh juice of Azhavanam Lawsonia inermis L.
(Fam. Lythraceae)
2. Katthazhai (Syn. Kariyabolam) Aloe barbadensis
Mill. Syn. Aloe vera Tourn. ex L. (Fam. Liliaceae) and
3. Sour skimmed milk of Cow (Bos taurus indicus)
2
Chukku (Dry Ginger) Zingiber
officinale Rosc. (Fam. Zingiberaceae)
Dried rhizome
Outer coat peeled off.
3
Milaku- Black Pepper Piper nigrum L.
(Fam. Piperaceae)
Fully mature dried fruit
Immature pin head sized fruits are thrown off. Roasted
slightly.
4
Thippili- Long pepper Piper longum
L. (Fam.Piperaceae)
Dried, immature, catkin-
like fruits with bracts
Immature pin head sized fruits are thrown off. Roasted
slightly.
5
Vasambu- Sweet flag Acorus calamus
Linn. (Fam. Araceae)
Dried rhizome
Washed and dried under shade.
Preparation of the test drug
100gms of purified Sulphur is taken. The ingredients of the chooranam are powdered separately and taken in the specified
ratio mentioned below and mixed well.
Dried ginger - 200gms
Pepper – 15gms
Long pepper- 15gms
Sweet flag- 15 gms
Half of the chooranam is taken in an earthern vessel above which the sulphur cake is placed. The remaining half of the
chooranam is put over and around the sulphur. The vessel is closed by another earthern vessel and made air tight by ‘Seelai seidhal’ in
which moist mud pasted ghada cloth (thick cotton cloth) is wrapped in 7 whorls at the rim of the two pots holding them tight. The
apparatus is subjected to ‘Pudam’ with 15 ‘varatties’ (cowdung cakes). Pudam is the process of heating in which the purified raw drug
or the pellet (for parpam or chenduram) or the apparatus which should undergo the heat (or burning) is placed in the centre and the
arrangement of cowdung cakes is done around it as if it is embedded within the specified fuel mentioned in that particular classical
text such that equal heat is rendered on all sides. The heating process is done in a pit. Care should be taken that the cakes should not
get burnt but should provide heat in a flame enough to provide constant heat and should not evolve much smoke too. The blackish
remnant obtained is collected, grinded to a fine powder and stored in air tight glass container. To be administered with Cow’s ghee.
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Analytical specification of Karuppu
Karuppu simulate centuram. So the analytical specfications are applicable for karuppu preparations and was done as per the
protocol for testing published by AYUSH[4].
Table.2 Analytical specifications for Karuppu.
S No
Analytical Specifications for Karuppu
1
Description, Colour, Odour
2
Identification
3
Particle size. Mesh size 200-300
4
Loss on drying at 105○ C
5
Total ash
6
Acid insoluble ash
7
Water soluble ash
8
Assay of element(s)
9
Assay of element(s)
10
Siddha specifications
Lustreless
Fine enough to enter the crevices of finger
Floats on water Smokeless
Tasteless
Irreversible
Organoleptic characteristics
Sensory parameters like colour, odour, taste, size of the particles were analyzed and recorded.
Physico Chemical Characterization
All the physicochemical parameters were carried out as per standard guidelines [5, 6]. Additionally pH was done at 25°C
(1:10 Ratio).
Determination of total ash
About 2 grams of the dried crude drug were weighed accurately in a tarred platinum or silica dish and was incinerated at a
temperature not exceeding 550oc until free from carbon. It was then cooled and weighed. The percentage of ash was calculated with
reference to air dried drug.
Determination of water soluble ash
The total ash was boiled for five minutes with 25ml of water. The insoluble matter was collected on a Gooch crucible (or) an
ash less filter paper. It was washed with hot water and ignited for 15 minutes, at a temperature not exceeding 550oc. The weight of the
insoluble matter was subtracted from the weight of the ash, the difference in the weight of the ash represents the water soluble ash.
The percentage of water soluble ash was calculated with reference to the dried drug.
Determination of acid insoluble ash
The total ash was boiled with 25ml of 2 M HCL for 15 minutes. The insoluble matter was collected on a Gooch crucible (or)
an ash less filter paper. It was washed with hot water and ignited. It was then cooled in a desiccators and weighed. The percentage of
acid insoluble ash was calculated with reference to the dried drug.
Loss on drying
5gm of the drug is heated in a hot oven at 40 degree C to constant weight. The percentage of loss of weight was calculated.
Qualitative analysis for presence of acid and basic radicals (Raman 2006)[7].
Preparation of extract
5gm of Gandaka karuppu is weighed accurately and placed in a 250ml clean beaker and added with 50 ml of distilled water.
It is then boiled well for about 20 minutes. Then it is covered and filtered in a 100ml volumetric flask and made up to 100 ml with
distilled water.
The aqueous extract was used for the analysis of acid radicals, basic radicals.
Test for acid radicals
Test for sulphate
2 ml of the above prepared extract is taken in a test tube. To this, 2 ml of 4 % ammonium oxalate solution is added. Cloudy
appearance/ white precipitate indicate the presence of sulphate.
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2ml of Sodium carbonate extract is added with 2 ml of dilute hydrochloric acid until the effervescence ceases off. 2ml of
Barium chloride solution is then added. A white precipitate insoluble in concentrated Hydrochloric acid confirm the presence of
sulphate.
Test for Chloride
2 ml of Sodium carbonate extract is added with dilute nitric acid till the effervescence ceases. Then 2 ml of silver nitrate
solution is added. Appearance of cloudy white precipitate completely soluble in excess of Ammonium hydroxide solution indicates the
presence of chloride.
Test for Phosphate
2ml of the extract is treated with 2 ml of ammonium molybdate solution and 2 ml of concentrated Nitric acid. Appearance of
yellow precipitate / cloudy appearance with yellow colour indicates the presence of phosphate.
Test for carbonates
2 ml of the extract is treated with 2 ml of magnesium sulphate solution. Cloudy appearance/white precipitate indicates the
presence of carbonates.
Test for Sulphide
1 gm of the substance is treated with 2 ml of concentrated Hydrochloric acid. Colourless rotten egg smelling gas turning lead
acetate paper black on warming indicates the presence of sulphide.
Test for Nitrate
1 gm of the substance is heated with copper turning and concentrated sulphuric acid and the test tube is viewed vertically
down. Copious evolution of reddish brown gas indicates the presence of nitrate
Test for Fluoride and Oxalate
2 ml of the extract is added with 2 ml of dilute acetic acid and 2 ml of calcium chloride solution and heated. Cloudy
appearance/ White precipitate indicate the presence of fluoride/oxalate.
5 drops of clear solution is added with 2ml of dilute sulphuric acid (H 2 SO 4) and slightly warmed. To this, 1 ml of dilute
potassium permanganate solution (KMnO4) is added. Decolourisation of Potassium permanganate solution indicates the presence of
oxalate.
Test for Nitrate
3 drops of the extract is placed on a filter paper. On that, 2 drops of acetic acid and 2 drops of benzidine solution is placed.
Development of Yellowish red colour confirm the presence of Nitrate.
Test for basic radicals
Test for Lead
2 ml of the extract is added with 2 ml of Potassium Iodide solution. Yellow precipitate soluble in hot water and reappearing
as “Golden Yellow fumes” on cooling indicates the presence of lead.
Test for copper
One pinch of substance is made into paste with concentrated Hydrochloric acid in a watch glass and introduced into the non-
luminous part of the flame. Appearance of bluish green coloured flame indicates the presence of copper.
2 ml of the extract is added with excess of Ammonia solution. Bluish precipitate or deep blue coloured solution indicates the
presence of copper.
Test for Aluminium
To the 2 ml of the extract Sodium hydroxide solution is added in drops to excess. Appearance of white precipitate soluble in
excess of Sodium Hydroxide indicates the presence of aluminium.
Test for Iron
To the 2ml of extract 2ml of Ammonium thiocyanate solution is added. Appearance of blood red colour indicates the
presence of Ferric iron.
To the 2 ml of extract 2 ml of Ammonium thiocyanate solution and 2 ml of concentrated Nitric Acid is added. Appearance of
blood red colour indicates the presence of Ferric iron.
Test for Zinc
To the 2 ml of extract, sodium hydroxide solution is added in drop of excess. Appearance of white precipitate solution in
excess of sodium hydroxide solution indicates the presence of zinc.
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Test for Calcium
2 ml of the extract is added with 2 ml of 4 % Ammonium oxalate solution. Cloudy appearance/white precipitate indicates the
presence of calcium.
Test for Magnesium
To the 2 ml of the extract sodium hydroxide solution is added in drops to excess. Appearance of white precipitate, insoluble
in excess of sodium hydroxide solution indicates the presence of magnesium.
Test for Ammonium
To 2 ml of extract few ml of Nessler’s reagent and excess of sodium hydroxide solution are added. Appearance of reddish
brown precipitate indicates the presence of ammonium.
Test for Potassium
A pinch of substance is treated with 2 ml of sodium nitrite solution, treated with 2 ml of cobalt nitrite in 30% glacial acetic
acid. Appearance of yellowish precipitate indicates the presence of potassium.
Test for Sodium
2 pinches of the substance is made into paste by using Hydrochloric acid and introduced into the blue flame. Appearance of
yellow colour flame indicates the presence of sodium.
Test for Mercury
2 ml of the extract is treated with 2 ml of Sodium hydroxide solution. Appearance of yellow precipitate indicates the presence
of Mercury.
Test for Arsenic
2 ml of extract is treated with 2 ml of silver nitrate solution. Yellow precipitate / brownish precipitate indicates the presence
of Arsenic.
Anti microbial study of Gandaka karuppu
Media used
Nutrient Agar per litre/gram (NA).
Peptone
-5.0
g
Yeast extract
-2.0
g
NaCl
-5.0
g
Agar
-18.0
g
Distilled water
-1000
ml
pH
– 7.0
Culture collection and maintenance
The organisms used for the study namely, E.Coli, Staphylococcus aureus, Salmonella typhii, Vibrio cholera, Pseudomonas
spp were obtained from CAS in Botany and Department of Biochemistry University of Madras, Guindy Campus, Chennai 600 025.
Preparation of culture for antibacterial studies
Twelve hours old bacterial suspension was adjusted to 0.5 OD and 1.0ml from the above was inoculated into 50 ml of
nutrient broth and incubated at 37 0 C in an orbital shaker at 150 rpm. To determine the growth rate, culture was removed at 4h
interval and the growth was monitored by measuring the optical density at 540nm in spectrophotometer. The growth curve was drawn
by plotting OD value against the incubation time.
Antibacterial susceptibility test[8]
Brain heart infusion (BHI) agar was used for susceptibility testing. Disks of 6mm in diameter were punched from a sheet of
Whatman filter paper, sterilized, and impregnated with 25 micro L each of 0.2 g/ml extract or solvent alone and dried at 30–35 degree
C for 12–24 h. The bacterial inoculums were prepared from subcultures of bacteria as follows: four to five colonies of the isolates
were emulsified in sterile distilled water and the turbidity adjusted to 1.5×108 CFU/ml (corresponding to 0.5 McFarland standards). A
sterile cotton swab was dipped into the standardized bacterial suspension and used to evenly inoculate the BHI agar plates. The plates
were allowed to dry for 3-5 min. Thereafter, all disks were placed on the plates and pressed gently to ensure complete contact with
agar. A distance of at least 15mm was maintained from the edges of the plates to prevent overlapping of inhibition zones. Fifteen
minutes following placement of the disks, the plates were incubated at 37◦C for 2–5 days. They were then examined and the diameter
of the zone of inhibition measured.
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Well-in agar method[9].
Anti-bacterial activity of aqueous extract of Gandaka karuppu was tested by a modified well-in agar method. The inoculum
suspension was spread uniformly over the agar plates using sterile cotton swab spreader, to get a uniform distribution of bacteria.
Subsequently, using a sterile borer, well of 0.5 cm diameter was made in the inoculated media. Different concentration drug 25, 50, 75
and 100 µg/ml of each extract was aseptically filled into the well. Later the plates were placed at room temperature for an hour to
allow diffusion of extract into the agar. Then the plates were incubated for 24 h at 37◦C. The results were recorded by measuring the
diameter of inhibition zone at the end of 24 h - 48 h.
RESULTS
Description
Fine powder and is similar to Centuram but differs from it, in being black in colour.
Table.3. Organoleptic Characters of Ingredients of Gandhaga Karuppu.
Ingredients
Characters of Crude drugs that complied with
SPI, API and others*
Characters of Obtained
Powdered ingredients
Sulphur
Yellow crystalline lumps with yellowish white
streaks, Resinous lusture, brittle, Transluscent
Yellow color, Characteristic Odor,
Bitter taste
Chukku
Laterally compressed Rhizome, branched on upper
side, each with a depressed scar at the apex
Cream color, and aromatic,
agreeable and pungent taste
Black
Pepper
Greyish-black to black, hard and wrinkled
Blackish-grey color, aromatic,
pungent taste
Thippili
Blackish brown or black, sessile, with small
protuberances of minute fruits that are evenly
arranged arranged around an axis; surface rough and
composite; Hard and brittle, fractured surface not
even but granular, shows a central axis and 6 to 12
fruitlets arranged around an axis.
Deep moss green color, pungent
taste producing numbness on the
tongue; odour aromatic.
Vasambu
Brownish-red or grayish-yellow externally, slightly
flattened, rough, branched, upper side marked with
alternately arranged, large,
Puff color, pungent and bitter taste.
broadly, triangular, transverse leaf scars which
almost encircle the rhizome, lower side shows
elevated tubercular spots of root scars, buff coloured
intemally, Hard and brittle, odour, aromatic; taste,
pungent and bitter.
SPI – Siddha Pharmacopoeia of India
API – Ayurvedic Pharmacopoeia of India
* Chinese Medicinal Material data base
Note – No significant difference is observed in the characteristic odour and taste of these crude drugs and respective powders after
purification and so these specific characters mentioned in the powder is also applicable to crude drugs.
Organoleptic characters of the finished product Gandhaga karuppu
Black in colour, characteristic odour is due to the ash of sulphur and very slight bitter taste. Complied with the Siddha
specifications for centuram. [Table.2.]
Physico chemical characterization
Table.3 Physico chemical analysis of Gandaga karuppu.
S. No
Physico chemical Parameters
Results
1
pH at 25°C (1:10 Ratio)
5.68
2
Ash Value @ 550°C (%)
16.40
3
Water soluble (%)
4.20
4
Alkalinity as CaCO3 in water soluble ash (%)
0.06
5
Acid Insoluble Ash, (%)
55.10
6
Loss of drying @ 105°C (%)
0.52
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Table .4 Qualitative chemical analysis of Gandaga karuppu.
S No
Acid radicals
Basic radicals
1
Sulphate
Iron
2
Chloride
Ammonium
3
Carbonate
4
Fluoride.
Anti microbial study of Gandaka Karuppu
The antibacterial activity of crude ethanol extract of Gandaka Karuppu was tested against various bacterial pathogens and
inhibition was determined by disc diffusion method.
Gandaka Karuppu showed no significant inhibition against the test pathogens. This may be due to the problem in the
solubility of the drug.
DISCUSSION
Karuppu in Siddha system of medicine is a ‘kajjali’(Ayurvedic dosage form) [10, 11] like black coloured powder. But in
Gandaka karuppu the black final product is obtained by the thermal reaction of sulphur with herbal ingredients without mercury. Other
karuppu medicines are Sivanar amirtham, Kasthuri karuppu, Pattu karuppu etc. Herbal Karukku medicines are Vasambu karukku,
Amai Odu Karukkuk Kudineer, Uthamani karukku etc. Gowri chindamani is a black coloured formulation of Siddha medicine made
of mercury and sulphur [12].
The organoleptic characters are the simple and quick evaluatory means for the establishment of identity and the quality and to
judge the material in its crude and powder form [13]. The results revealed conceals the fact that the crude drugs used for preparation of
formulation lie within the limit which signifies their good quality and purity. The shelf life and stability of a drug depend on the
deterioration time and in turn on the amount of water present in the formulation. The higher moisture content deteriorate the
formulation due to fungus. Insufficient drying favors the spoilage by molds and bacteria and makes possible the enzymatic destruction
of active principles. Apart from the dryness of the drug, the moisture removal rate is also equally important together with the removal
condition. If the rate is too slow, much spoilage may occur before the drying process is completed [14]. The loss on drying 0.52 % in
the final product signify that the drugs are properly dried and stored. The shelf life for Karuppu is generally 1 year as per Siddha
formulary [15] but recently notified to be 2 and 5 yrs based on the sources of ingredients. Since Gandaga karuppu is a herbo mineral
drug, the shelf life is 5 yrs amendment [16].
The pH conventionally represents the acidity and alkalinity [17]. The pH value is influences the quality of medicine [18] and
it controls many chemical and microbiological reactions [19]. Under low pH value (presence of acidic substances), the bacterial count
could be low, but at neutral or higher pH, contamination of the herbal preparations could observed to be higher. This suggests that a
neutral or alkaline pH favoured high contamination levels of the herbal preparations [20]. The pH obtained 5.7 indicates that the drug
is acidic. This pH from mineral or phytochemicals is emphazised to be evaluated so that clarity regarding the consideration of sulphur
alone in the final product or inclusion of the charred churanam part too in the final product, be achieved. The results obtained in this
standardization study included the churanam part. The potency is also to be considered regarding fixing up of the final product. Fresh
ash leachates of volcanic ash which is high in sulphur (comparison is justified below) are found readily soluble and pH varied highly.
However, the ash from the later phase of the eruption was more acidic, with a pH of approximately 5.1. The contribution to the pH
obtained in the drug and contribution to the water soluble extractive from the percentage of minerals other than the primary ingredient
sulphur and from the phytochemicals need to be explored and compared. If the contributing factor is mineral the shelf life would be as
expected and there would be meagre chances of immediate deterioration. Otherwise, if from phytochemicals (oxygenated compounds)
then chances of contamination is comparatively higher. The importance of extent of care during storage of Gandaka karuppu is dealt
here based on pH.
Ashing involves an oxidation of the components of the product. A high ash value is indicative of contamination, substitution,
adulteration or carelessness in preparing the crude drug for marketing [17, 21]. The total ash usually consists of carbonates,
phosphates, silicates and silica which include both physiological ash (plant tissue) and non-physiological ash (environmental
contamination such as sand and soil) [17, 22, and 23]. The total ash observed in the siddha drug is 16.40%. Physiological ash
interferes with the illustration of total ash in reflecting the quality of herbal medicines e.g. Calcium oxalate. Thus the Acid insoluble
ash also establish the quality of Siddha herbal formulation. The high Acid insoluble ash might be due to the addition of Sulphur and
the other minerals in other ingredients of the formulation. The water-soluble extractive value denotes the presence of phytochemicals
such as sugar & acids and inorganic compounds. The water soluble extractive value in the present Siddha compound formulation
found be 4.2% indicated some water soluble components in the formulation. Clinically the drug showed better improvement for the
treatment of the Skin disease (unpublished data and in experience). The acid radicals present were sulphate, chloride, carbonate,
fluoride. This result is in agreement with all the major species of acid radicals detected in fresh ash leachates of volcanic ash (Witham
et al., 2006) [24] and carbonates from phyto components of the drug. The basic radicals present were iron and ammonium. The
volcanic ash is high in sulphur content and is of traditional and pharmaceutical importance that Volcanic Earth Healing Centre Spa
(http://www.volcanicearth.com) emerged and uses the ash extensively and quotes that the Sulphur in Volcanic Ash is a proven, safe,
age-old remedy that has been used to treat every conceivable skin irritation and infection.
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Transformation of sulphur to biosulphur during the heat processing of drug with other herbal ingredients(pudam) to render
the elemental sulphur to be partly hydrophilic need to be analyzed in forthcoming studies. This hydrophilic nature might be
responsible for the efficacious activity of the drug clinically as indicated in the classical text for skin diseases. More than the direct
solubility, the rate of solution, dissolution and the respective pH would provide predictive results in future to know the fate of purified
and processed sulphur through Siddha traditional methods, in the drug in case of preliminary tests.
Standardization ensure the safety of drugs through limit tests for heavy metals. Though this study does not include the
parameter, some sulphur based literary siddha research evidences support the safety of the drug GK and is proven to be low toxic [12].
Poorṇa Cantirotaya Centuram is prepared with gold, mercury, and sulphur (major ingredient). The drug is reported that it can be
classified as Category 5 with low acute toxicity hazard. In both acute and repeated dose toxicity studies, the drug showed no
significant adverse effects. The research evidences regarding the proven safety of the Siddha formulation Rasagandhi Mezhugu
(RGM), with sulphur as one of the ingredients is documented [25]. These sulphur based safety of non- karuppu dosage forms support
the safety of Gandhaka karuppu too. In specific acute toxicity of Gandaga karuppu has proved to be non toxic (D. Deepa Bai., et al –
to be published soon). Vasambu karukku is a herbal karuppu paediatric dosage form[26]. This karukku is the purified form of the drug
[27].
Inspite of improvements in science, identification of the potentcy is still based on traditional methods [28]. The indication of
the drug GK, recommended to be administered with the melted cow’s ghee is either to disperse the supposed nano particulate drug by
hot melt method [29] or improve the miscibility to enhance the acceptability and efficacy of the drug. The cow’s ghee which serve
both as a vehicle and an adjuvant guide to expect that the constituents in the final product to be thermo-stable in the melted form of
ghee. The role of supercritical CO2 and the necessary conditions that fit in the processing of Gandaka Karuppu, are discussed as the
solubility enhancer [30] and efficacy enhancer in the drug (Deepa Bai et al- to be published soon). The sustained availability of
genuine raw materials improves the quality and effectiveness of traditional medicines in the present world of cost effectiveness,
adulterated, substituted and controversial drugs [31]. The ingredients of GK Siddha medicine are commonly available at reliable cost.
In specific, a comparative TLC of the drug with that of the ‘Thin Layer Chromatographic Atlas of Ayurvedic Pharmacopoeial
Drugs’ for piper nigrum[32] and comparative TLC of other herbal ingredients from other pharmacopoeias would be an updated study
and an immediate requirement to complete the preliminary standardization of Gandaka karuppu.
CONCLUSION
The physico-chemical standardization authenticate the finished Siddha herbo mineral product to be of acceptable and
standard quality and is devoid of adulterants. The results of this study may be used as the reference standard in further research
undertakings of its kind and thus fulfilling the scope of Knowledge based medicinal products (Dr. Handa’s statement)[33].
Future research
Standardization and quality control with integrated traditional knowledge and modern scientific techniques is important [34].
Since this is the first documentation on the drug Gandhaga karuppu for its authenticity, advanced analytical techniques like 1. SEM
(Scanning Electron Microscopy), AFM (Atomic Force Microscopy) studies are needed to validate the fact involved in solubility/
adsorptive capability of sulphur 2. Preliminary sieving tests and particle size analyzation and compare the grinding time of karuppu
and confirm the micronisation, 3. X- ray Diffraction studies to identify the crystalline phases in the processed sulphur and functional
group identification techniques like IR and NMR [35] with an expectation for the functional groups to cleave the disulphide bonds of
hyperkeratinised cells and improve this Siddha medicine as scientifically validated potent drug for the treating the skin diseases and
for other indications mentioned in the classical text.
The processed and supposed bio-sulphur may be tried to use it commercially as biofuel, bio-fertilizer. AAS (Atomic
absorption studies) are also essential for the confirmation of percentage of minerals. Phytochemical studies, assay of active ingredients
in Gandhaga karuppu, quantification through HPLC and HPTLC [36] are further required.
ACKNOWLEDGEMENTS
I am highly privileged to acknowledge with thanks to Prof. Dr. Ramaswamy, DG, CCRS Chennai-106 for their
encouragement. I express my thanks to my Supervisor Dr. M. Allimuthu M.D (Siddha), former Head of the Department of
Gunapadam Branch, for his valuable guidance throughout the study. I also thank to Dr. Jayanthi Athikkal (PSO-Phg.), PCIM&H,
Ghaziabad and Dr. Sathyarajeswaran parameswaran Director, i/c, SCRI, DR. Shyamala Raj kumar(RO),CCRS, for their extended kind
support. I also thank Mr. Rajneesh Kumar, Dr. Suda Revathy Sendilkumar and Mr. Sendilkumar for their timely help. I thank all the
staff members of both the institutes for their cooperation.
I gratefully acknowledge my sincere thanks to The TN Dr. MGR Medical University, Guindy, Chennai - 600032 and our
former principal Dr. Khadhar, Govt Siddha Medical College, Chennai-106.
Declaration
There is no conflict of interest.
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