Analytical Study of
Thalangi Ennai: A Pain Relieving
Siddha Medicated Oil
Vinotha Sanmugarajah, Ira Thabrew,
Sri Ranjani Sivapalan
Unit of Siddha Medicine, University of Jaffna, Sri Lanka
Institute of Biochemistry, Molecular Biology and Bio
technology, University of Colombo, Sri Lanka
1
Introduction
Siddha, Ayurveda and Unani are the three major
Traditional Systems of Medicine in Sri Lanka.
Siddha which is largely therapeutic in nature has a
rich heritage and history.
The word ‘Siddha’ means ‘Perfection’ and those who
attained perfection were known as ‘Siddhars’ the
pioneers of the system.
2
The Siddhars were great scientists in ancient times.
Siddha is widely practiced in the Tamil speaking
areas of Northern and Eastern provinces.
According to tradition, the origin of Siddha system
of Medicine is attributed to the great Siddha
Agastiyar.
Some of his works are still standard books of
Medicine and surgery in daily use among the Siddha
Medical practitioners and Traditional healers.
3
Siddha Materia Medica
The resources of Siddha Materia Medica are
classified in to three major divisions namely,
Herbal
Metals and Mineral
Animal kingdoms
According to Siddha literatures, there are 64 dosage
forms of which 32 are meant for oral (internal) route
and 32 for parental & external (including topical)
applications.
4
Effective management of vātā disorders such as
rheumatic pain, bone degenerative changes and
impaired movement of limbs are the important tasks
in the field of treatment.
Siddhars described vātā disorders of different origin
and external application is most useful for their
management.
Among them Ennai (Medicated oil) is one of the
medicaments prescribed by them.
5
Thalangai ennai is one of the Siddha medicated oil
mentioned in the Siddha Pharmacopeia and it is
prepared at Siddha Drug Unit in Jaffna and supplied
to all Government District Ayurvedic Hospital, Rural
Ayurvedic Hospitals and Central Ayurvedic
Dispensaries in Jaffna District.
This oil is showing clinically enhanced pain relieving
properties which requires analytical study for
standardization.
6
Botanically pure and authentic ingredients were used
in the preparation of T. ennai.
Ingredients
It is a compound medicine and its ingredients are,
Juice of the thalangai (Pandanus tectorius) -1250 ml
Sesame oil - 625 ml
Cow’s milk - 625 ml
Water - 16000 ml
Rock salt -10 g
Seventeen different herbal raw materials - each 10 g
7
Details of ingredients of Thalangai ennai
No Local name
Asclepiadaceae
Labiatae
Refrigent
Cold
Gramineae
Stimulant
Refrigent
Analgesic
Cold
5
6
Hemidesmus
indicus
Villamichu
Plectranthus
ver
zeylanicus
Vetti ver
Andropogon
muricatus
Sittamutti
Pavonia
zeylanica
Peramutti ver Pavonia odorata
Tevadaru
Cedrus deodara
Pharmacological Potency
action
Analgesic
Cold
Cold
Hot
7
Jatamashi
Velerianaceae
Analgesic
Anti
inflammatory
Aromatic
1
2
3
4
8
9
Scientific name Family name
Nannari ver
Nardostachys
jatamansi
Kacholum
kaempferia
galanga
Mara manjal Coscinum
fenestratum
Malvaceae
Malvaceae
Coniferae
Zingiberaceae
Aromatic
Stimulant
Menispermaceae Analgesic
Aromatic
Cold
Hot
Hot
Hot and
Cold 8
10
11
12
13
Sandanam
Kottam
Kundurukkan
Adi-maduram
14 Satakuppi
15 Chirttilam/
elakaya
16 Patchchilai
17 Koraikizangu
18 Thazhai/
thazaai
Santalum album
Costus speciosus
Boswellia serrata
Glycyrrhizae glabra
Santalaceae
Scitaminaceae
Burseraceae
Papilionaceae
Peucedanum
graveolens
Eletteria
cardamomum
Zanthochymus
piotorlins
Cyperus rotundus
Umbelliferae
Zingiberaceae
Cyperaceae
19 Rock salt
20 Sesame oil
Pandanus tectorius
Pandanaceae
(Screw pine/ umbrella (Mudu- keyiya)
tree: Ketaki/ ketaka)
NaCl impura
Sesamum indicum
Pedaliaceae
21 Cow’s milk
Bos indicus inn
22 Water
Analgesic
Stimulant
Stimulant
Emmolient
Analgesic
Deobstruent
Stimulant
Stimulant
Aromatic
Stimulant
Aromatic
Stimulant
Cold
Hot
Hot
Cold
Refrigent
Aromatic
Antispasmodic
Stimulant
Analgesic
Cold
Hot
Hot
Hot
Cold
Hot
Hot
9
Preparation Method of T. ennai
From 1 to 17 dried raw materials are ground to get
coarse powder and mixed with water. Then
continued boiling to reduce the volume to one
eighth (2000 ml).
Juice of the thalangai, sesame oil, milk (2:1:1) are
taken in a vessel and heated for some time; mixed
the aqueous decoction together. This mixture is
boiled on mild fire with stirring and boiling
continued till all the water evaporates and this
mixture turn as oily (waxy like). After cooled 10
minutes, the powdered rock salt mixed with the oil.
10
Objective
The aim of this study was to evaluate the
scientific validation of the Thalangai ennai.
11
Methodology
For the scientific validation of this oil,
Physicochemical analysis such as pH value,
specific gravity and organoleptic characters such as
colour, odour, appearance, touch and clarity were
performed.
Thin Layer Chromatography (TLC) finger print of
the Thalangai ennai was studied after dissolved the
oil in water and extracted with dichloromethane.
The oil sample was purchased from Siddha Drug
Unit, Atchuvely, Jaffna.
12
Physicochemical evaluation
Sample of oil (Thalangai ennai) were subjected for
determination of physicochemical parameters such as
organoleptic characters, pH value and specific
gravity.
Organoleptic evalution
These refer to macroscopic identification of the
formulation.
Sample of oil (Thalangai ennai), water and
dichoromethane extracts of oil were subjected to the
organoleptic characterization such as appearance,
touch, colour, clarity and odour.
13
Determination of pH range
The pH of sample of oil (Thalangai ennai) was
determined using standard simple glass electrode
pH meter (Consort). The oil was weighed to about
40ml in separate beaker and the pH of the oil was
determined using a pH meter at room temperature
(29 0C).
14
Determination of specific gravity
(http://www.engineeringcivil.com)
Clean, dry and weighed the specific gravity bottle
along with the stopper (Weight ‘A’). Then filled the
specific gravity bottle with freshly distilled water
and inserted the stopper firmly. Kept it in a room
temperature and weighed it (Weight ‘B’). Finally,
filled the specific gravity bottle with the oil sample
and weighed separately (Weight ‘C’).
Specific gravity (oil) = (C – A)
(B – A)
15
Development of TLC fingerprints and standardization
of thalangai ennai
TLC finger print of the T.ennai was studied after
dissolving the oil in water and extracted with
dichloromethane.
Prior to testing, prepared a water and
dichloromethane extract of the oil to examined, used
a rapid extraction process, as followed.
Place about 50 ml thalangai ennai and fume ships of
in an accurately weighed, glass-stoppered round
bottom flask.
16
For the water extraction, 20 ml water was added to
the flask and weighed to obtain the total weight
including the flask.
Then extraction was done by using reflux condenser
for 2 hours on heating mantle (Electro thermal). Then
cooled and transferred to the Separator funnel system.
After 20 minutes, the water extract was separated in
to the clean small flask and kept tightly closed with
lid. Further 20 ml water added to the residue in the
round bottom flask and water extraction was done
again as the same procedure.
17
After that, the whole separated water extract (30 ml)
transferred in to the separator funnel system.
Then 10 ml dichloromethane added to the water
extract and mixed well. After 20 minutes, the
dichloromethane extract was separated in to the
accurately weighed, small round bottom flask and
kept tightly closed with lid.
Further 10 ml dichloromethane added to the residue
in the separator funnel system and dichloromethane
extract was separated again as the same procedure.
18
Then this procedure repeated again. Totally,
dichloromethane extract separation was done for
three times.
Whole dichloromethane extract collected in to the
round bottom flask and evaporated on a Rotary
evaporator (Buchi) for just as long as was required
to remove the solvent, and re-dissolved the residue
in a small volume of dichloromethane (3 ml).
Finally, the whole extract was collected in clean
stoppered glass test tube and used for spotting the
chromatographic plates.
19
Five µl of oil extract was spotted on to TLC plates
(8.5 x 5.3 cm) coated with silica gel (pre-coated,
GF254) and separated using different solvent
systems.
After development visible spots were not found for
oil extract. Numbers of spots were observed under
UV light (254 & 366 nm).
Visualization was attempted by spraying with
vanillin sulphate reagent for oil extract and heating
the plate for 5-10 minute (100-105°C).
20
The colour and Rf values of the spots were recorded
carefully and the chromatogram was documented by
graphical copying.
(Rf value = The distance between the point of
application and the centre of the spot of the material
being examined / The distance between the point of
application and the solvent front)
21
Results and Discussion
The physicochemical standards available for the
standardization of siddha oils are insufficient.
The analytical values available in the
pharmacopoeia standards for siddha formulations
are not finger print standards for each oil.
The first step towards this goal, Thalangai ennai
was subjected to systematic physicochemical and
Thin layer chromatography (TLC) analysis to
determine the quality and purity of that oil.
22
Table No. 1: Analytical parameters of Thalangai Ennai
Parameters
Appearance
Thalangai Ennai
Viscous
Touch
Oily
Colour
Clarity
Golden brown
Clear
Odour
Characteristic
pH value (290C)
4.61
Specific gravity
0.9185
23
Table No.2: Organoleptic properties of the water and
dichloromethane extracts of the Thalangai Ennai
Parameters
Appearance
Thalangai Ennai
Water extract
Dichloromethane extract
Liquid
Liquid
Touch
Liquid
Liquid
Colour
Light yellow
yellow
Clarity
Clear
Clear
Odour
Characteristic
Characteristic
24
Thalangai Ennai of Government
Siddha Drug Unit
Dichloromethane extract of
Thalangai ennai
25
According to Table 1, The results showed that this
oil (T. ennai) contains specific gravity 0.9185 and
pH value 4.61 at room temperature (290C).
As seen in Table 2, both extracts of the oil (T. ennai)
had similar organoleptic properties except for the
colour of the water and dichloromethane extracts.
26
254 nm
366 nm
Visible after spray
TLC finger print analysis of extract of Thalangai Ennai
27
Table 3: TLC finger print analysis of extract of Thalangai Ennai
Solvent system:- Pure Dichloromethane (volume)
Solvent front:- 7 cm
Rf values and colour of spots {*- intense, $- Moderately intense, #- Faint}
0.0286 * Brown
0.4000$ Pink
0.0571$ Pink
0.4428$ Green
0.0714* Brown
0.5286# Pink
0.1143# Grey
0.5857# Green
0.1857# Brown
0.8571# Pink
0.2286$ Pink
0.9143* Orange
0.2571$ Grey
0.9714* Pink
0.3571# Pink
28
Table 3, summarizes the Rf values and colour of
spots visible in the TLC profiles of the water and
dichloromethane extract of Thalangai ennai in Pure
Dichloromethane solvent system.
Although the separation of the extracts occurred in
the solvent system comprised of hexane:
dichloromethane: 1% methanol in dichloromethane
(1: 4: 5 v/ v) as the mobile phase, the best separation
of the extracts occurred in the solvent system
comprised of pure dichloromethane (v) as the
mobile phase.
29
The TLC of finger print of the oil sample showed
fifteen spots with Rf values ranging from 0.0286 to
0.9714 in the pure dichloromethane solvent system.
30
Conclusion
As there is not any evidence for detailed
physicochemical and TLC evaluation on
Thalangai ennai is reported.
Therefore present work is taken up in the view
to standardize the Thalangai ennai.
In the present study Thalangai ennai was
subjected to physicochemical parameters and
TLC finger printing for standardization.
31
Although, these preliminary physicochemical and the
TLC standards presented in this study can be used as
finger print standards for Thalangai ennai, further
research work of TLC studies of each single
ingredient of T. ennai should be carried out in future.
32
Acknowledgement
The financial support provided by the Higher
Education Twenty-first Century (HETC) Project,
Ministry of Higher Education, Nugegoda, Sri Lanka
(Reference No - JFN/ Sidda/ N1)
The guidance given by Dr. (Mrs.) Menuka
Arawwawala, Industrial Technology Institute (ITI),
Colombo 07, to conduct the physicochemical
studies are gratefully acknowledged.
33
THANK YOU
34
35
254 nm
366 nm
Visible after spray
TLC finger print analysis of extract of Thalangai Ennai
36