Ayurvedic formulations contain minerals, metals and toxic plant materials which are harmful to the human body. When these drugs are used in Ayurvedic formulations they should be purified to eliminate such impurities or toxic substances. But sometimes commonly used drugs show Adverse Drug Reactions (ADR). This may be because of the wrong Shodhana procedure adopted or improper Shodhana. Ayurveda has also given prime importance to Mahavishas and Upavishas and have used them in various Chikitsa. So at this stage it becomes mandatory to compare the effect of different media and processes on chemical constituents after Shodhana processes and validate the best Shodhana media/processes for that particular toxic substance. So the present study - “Phytochemical and Physico-chemical analysis of Jayapala Beeja (croton tiglium linn.) with reference to different Shodhana Samskara.” was carried out to show the best suitable medium for it’s purification.

1. “PHYTOCHEMICAL AND PHYSICO-CHEMICAL
ANALYSIS OF JAYAPALA BEEJA (CROTON TIGLIUM
LINN.) WITH REFERENCE TO DIFFERENT SHODHANA
SAMSKARA”
Dr. Jyoti M. Ganer. (Baragi)
Assistant Professor
Dept. of Agada Tantra Evam Vyavahara
Ayurveda

2. Ayurvedic formulations contain minerals, metals and toxic
plant materials which are harmful to the human body.
When these drugs are used in Ayurvedic formulations they
should be purified to eliminate such impurities or toxic
substances.
It is very interesting to observe that different medias and
processes have been explained for the Shodhana of a
particular drug.
But sometimes commonly used drugs show Adverse Drug
Reactions (ADR). This may be because of the wrong
Shodhana procedure adopted or improper Shodhana.

3. Ayurveda has also given prime importance to Mahavishas
and Upavishas and have used them in various Chikitsa.
So at this stage it becomes mandatory to compare the
effect of different media and processes on chemical
constituents after Shodhana processes and validate the
best Shodhana media/processes for that particular toxic
substance.
So the present study - “Phytochemical and Physico-
chemical analysis of Jayapala Beeja (croton tiglium linn.)
with reference to different Shodhana Samskara.” was
carried out to show the best suitable medium for it’s
purification.

4. • Till date only one work has been carried out on
Jayapala Shodhana by applying only one Shodhana
process of Svedana with Godugdha.
• Dr. Arthi Shinde - Study of effect of Shodana Process
on LD50 of Croton tiglium (Jayapala Beeja), Tilak
Ayurved Mahavidyalaya. Pune – 2005.

5. Objectives of the Study
To compile the classical and recent literature
concerned with the drug - Jayapala.
To carry out Shodhana (purification) of Jayapala seeds
by different 6 methods mentioned in the classics.
To compare the phyto-chemical and physico-chemical
study of Ashodhita Jayapala Beeja with Shodhita
samples of Jayapala Beeja.
To compare the percentage of toxic principle of various
processes.

6. Review on
Visha
Concept of
Shodhana
Review of
Drug
Jayapala
Pharmacog-
nostical
Study
Pharmaceut
-ical Study
Analytical
Study
Discussion
&
Conclusion

7. The literary study has been carried out in 3 phases.
Concept of Visha: In this phase brief review regarding
Visha, Upavisha and its classification mentioned in different
classics of Ayurveda has been compiled and interpreted
accordingly.
Concept of Shodhana: In this phase the concept regarding
Shodhana procedure mentioned in different classics of
Ayurveda has been compiled and interpreted accordingly.
Drug review: In this phase, review of various Ayurvedic and
modern literatures and published papers have been carried
out to gather information about the drugs used in this study.
i.e. Jayapala, Godugda, Tankana, Nimbu, Mahisha Mala and
Gomaya.

8. Jayapala
Latin Name : Croton tiglium Linn.
Family : Euphorbiaceae
Rasa : Tikta (Su.Su.42/22).
Guna : Guru, Ruksha, Tikshna
Virya : Ushna
Vipaka : Katu
Doshaghnata : Kaphapitta Shamaka
Karma: Virechana, Lekhana, Vidahi, Svedajana, Dipana,
Krimighna, Shothahara, Jvaraghna, Vishaghna.
Rogaghnata : Jirna Vibandha, Agnimandya, Krimi,
Jalodara, Mastiskagata Raktasrava, Sanyasa,
Sarvangashotha, Jvara, Jangama Visha, Sarpa Visha.

9. Chemical constituents:
Seeds contain stable oil - 30 to 40%.
The active part of Croton is Crotin a toxalbumin.
Poisoning by Croton Oil:
Croton oil is a fixed oil, extracted by pressure from the
seeds of Croton tiglium. It is used in medicine as a
powerful purgative, in doses of one or two drops. In
overdoses, it acts as a violent irritant to the gastro-
intestinal mucous membrane, causes excessive vomiting
and purging, followed by collapse.
Fatal Dose:
Children : One seed or 3 drops of oil, Adults : Four seeds
or 20 drops of oil
Fatal Period: 4 to 5 hours and a maximum of 3 days to 6
days.

10. Medico Legal Importance:
Oil is ingested for suicidal purpose though rarely.
Abortifacient.
Administered with homicidal intent.
Arrow poisoning
Accidental poisoning due to therapeutic misuse.
Post Mortem Appearance:
Red, inflamed mucous membrane of stomach and
intestines.
Excoriated at places
Congested liver and spleen
Swollen and congested kidney
Occasionally post mortem findings are negative.

11. 1. Collection and identification of Jayapala Beeja: Raw
sample of Jayapala was collected from a local herbal drug
dealer and identified from the experts of the department of
Dravya guna.
2. Pharmagognostical study: Authentificaton of the drug
Jayapala was done at AYUSH approved research lab.
3. Pharmaceutical Study: In this phase, six different
methods of Jayapala Shodhana was done.
4. Analytical Study: Ashodhita Jayapala seeds and Shodhita
Jayapala seeds obtained by six different methods of
Shodhana were comparatively analyzed for any differences
in Organoleptic, Physico-chemical and Phyto-chemical
parameters.

12. Objective: To standardize the
Jayapala Beeja used for the
pharmaceutical process.
Here, the drug Croton tiglium
was authenticated
morphologically, macro and
microscopically.
Result : The given sample
was authenticated as seeds of
Croton tiglium as the macro-
microscopic features
mentioned for Jayapala in API
matches with that of test
sample.

13. Objectives of Pharmaceutical Study:
1. To carry out Shodhana (purification) of Jayapala seeds
by different methods mentioned in the classics.
2. To compare the effect of different media on toxic
principles.
3. To find out a best and suitable Shodhana process for
Jayapala.
Procurement of Raw Drugs:
Ashodhita Jayapala Seeds were collected from a local
market. Godugda, Gomaya, Mahisha Mala were taken
from a local Goshala. Shodhita Tankana was taken from
the pharmacy of the college and Nimbu (lime) were bought
from the local market.

14. Ref. of Shodhana Process Method
Rasatarangini (24/310-312) A
Swedana in Godugda (1 prahara
/3 times)
Rasatarangini (24/313-314) B
Swedana in Godugda with
Tankana
Rasatarangini (24/315-317) C Swedana in Godugda (1 prahara)
Rasamruta D
Swedana in Godugda & Bhavana
with Nimbu Rasa.
Yogaratnakara E
Sthapana in Mahisha Mala &
Bhavana with Nimbu Rasa.
Yogaratnakara F Swedana in Gomaya Rasa.

15. 1st group of Jayapala endosperm were subjected to
Shodhana by process of Svedana with Godugdha for 1
Prahara (3 hours) for 3 times.
Probable mode of action of the process:
The Swedana procedure may be considered as a hot
continuous extraction process where the toxic principles of
the plant are extracted.
Here Swedana was done in Godugda which possesses ten
properties opposite of the Visha. So according to Samanya
Vishesha Siddhanta, Dugdha can pacify the effect of Visha.
Dugda has Madhura Rasa, Madhura Vipaka and Sheeta
Virya, those are also opposite to that of Jayapala i.e. Katu,
Tikta Rasa, Katu Vipaka and Ushna Virya.
1st method of Shodhana

16. 2nd group of Jayapala endosperm were subjected to
Shodhana by mixing with 1/8th part of Tankana and
processed by Svedana with Godugdha for 2 Prahara (6
hours).
Probable mode of action of the process:
Tankana is included in Shodhaniya and Dravaka Gana and it
is used for Shodhana of different drugs and metals. Due to
adsorbing property, Tankana may absorb the poison from the
drug, and so is used for management of the cases of
poisoning mainly due to Sthavara Visha. Borax is also used
as an antidote in fluoride intoxication.
2nd method of Shodhana

17. 3rd group of Jayapala endosperm were subjected to
Shodhana by process of Svedana with Godugdha for 1
Prahara (3 hours) then the Jayapala endosperm were
collected from Pottali and washed with hot water and made
into paste and the paste was smeared on new Sharava
(earthen sacker) and kept for 1- 2 days for drying the oil
portion and then collected and dried in sunlight.
Probable mode of action of the process:
After the Swedana process the paste of endosperm was
smeared over the new earthen plate (Sharava). The object
of this process is, to absorb maximum oil from the paste of
the Jayapala endosperm. The use of new earthen pot is
suggested due to its adsorbent capacity.
3rd method of Shodhana

18. 4th group of Jayapala endosperm were subjected to
Swedana in Dola Yantra with Godugda for 1 Prahara (3
hours) then the Jayapala endosperm was collected from
Pottali and dried and powdered. Bhavana with Nimbu
Svarasa was given for three days, and then sun dried and
used.
Probable mode of action of the process:
During rubbing between surfaces of mortar and pestle
some heat is produced, so there may be possibility of
occurrence of chemical reaction in between materials and
media or in between materials.
Nimbu Swarasa (pH 2.0) being acidic in nature may
facilitate the extraction and evaporation of crotin a
toxalbumin.
4th method of Shodhana

19. 5th group of Jayapala endosperm were kept in Mahisha
Mala for 3 days and then removed and washed in hot water
and made into paste and the paste was smeared on new
Sarava (earthen sacker) and kept for 1- 2 days for drying
the oil portion and then Bhavana with Nimbu Svarasa was
given for three days, and then sun dried and used.
Probable mode of action of the process:
Sthapana means keeping the drug soaked in the liquid
medium. Here the outer skin of Jayapala is removed with
this treatment and it will loosen the toxic ingredients of the
Jayapala, which again are removed by washing the
endosperm with hot water and then smearing it on the
earthern pot and the Bhavana process with the Nimbu
Swarasa also helps to remove the toxic principles of the
drug Jayapala.
5th method of Shodhana

20. 6th group of Jayapala endosperm were subjected to
Shodhana by process of Svedana with Gomaya Rasa for 1
Prahara (3 hours).
Probable mode of action of the process:
Gomaya is used as a liquid media for the Svedana
Samskara. Here Gomaya is acidic in nature and it may
facilitate some chemical reaction with the drug. Principles of
Svedana method help in the extraction of the poison, where
the solvent enters the cells resulting in the swelling of
tissues making easy escape of the soluble constituent.
6th method of Shodhana

22. Objective:
To compare the Phyto-chemical and Physico-chemical study
of Ashodhita Jayapala Beeja and Shodhita samples of
Jayapala Beeja.
Plan of the Study:
Ashodhita Jayapala seeds and Shodhita Jayapala seeds
obtained by six different methods of Shodhana will be
comparatively analyzed for any differences in Physico-
chemical parameters and phyto-chemical study with
following parameters,

23. Organoleptic characters like Colour (Rupa), Odour
(Gandha), Touch (Sparsha), Taste (Rasa).
Foreign matter (Ashodhita Jayapala)
Loss on drying at 110°C
pH of 5% Aqueous Soln.
Ash value
Acid insoluble ash
Water soluble extractive value
Alcohol soluble extractive value
Quantitative estimation of Croton oil
Thin Layer Chromatography
High Power Thin Layer Chromatography: of all 7
samples (Ashodhita samples and 6 Shodhita samples)

25. Organoleptic characteristics of various sample of
Jayapala.
Sr.No.
Organoleptic
characteristics
Sample-RW
Sample-A
Sample-B
Sample-C
Sample-D
Sample-E
Sample-F
1 Color White Dull white Dull white Dull white Dull white
Grayish
white
Grayish
white
2 Odour
Charact
eristic
Characteris
tic - milk
Characteris
tic - milk
Characteris
tic - milk
Characteri
stic -
lemon
Characte
ristic -
lemon
Characte
ristic -
dung
3 Touch Soft Soft Soft Soft Soft Soft Soft
4 Taste -----
Sweet
pungent
Sweet
pungent
Sweet
pungent
Sour Sour
Salty
pungent

26. Physico-chemical tests of Raw and Shodhita
Jayapala.
Parameters
Sample
RW A B C D E F
Foreign matter (%) 6.8 NA NA NA NA NA NA
pH 5% Aqueous Soln
(%w/v)
6.03 6.36 5.13 6.24 4.35 5.33 5.85
Loss on drying at 105°C
(% w/w)
3.92 1.78 6.75 1.81 6.02 3.71 2.55
Ash value (% w/w) 2.80 2.74 4.96 2.61 2.37 2.99 2.68
Acid insoluble ash (%
w/w)
0.50 0.48 0.51 0.48 0.47 0.53 0.44
Water soluble extractive
(% w/w)
7.64 1.88 4.44 1.42 3.37 8.44 1.52
Alcohol soluble
extractive (% w/w)
29.16 22.58 23.88 15.37 24.78 22.64 16.83

27. Quantitative estimation of Croton oil in Raw and
Shodhita Jayapala
Parameters
Sample
RW A B C D E F
Fixed oil (%) 50.77 49.36 18.04 49.98 48.66 49.29 49.39
0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
60.00%
Raw A B C D E F
50.77% 49.36%
18.04%
49.98% 48.66% 49.29% 49.39%
Croton Oil

28. Rf values of TLC at 254nm.
TLC
analysis
Sample No. of Spots Rf values
Under UV
254nm
RW 6 spots
0.05 (D. green), 0.07 (D. green), 0.11 (D.
green), 0.45 (L. green), 0.57 (D. green), 0.65
(D. green).
A 7 spots
0.03 (D. green), 0.07 (D. green), 0.27 (L.
green), 0.45 (D. green), 0.57 (D. green), 0.65
(D. green), 0.83 (L. green).
B 2 spots 0.56 (L. green), 0.65 (D. green).
C 3 spots
0.07 (D. green), 0.56 (L. green), 0.65 (L.
green).
D 6 spots
0.03 (D. green), 0.05 (D. green), 0.09 (L.
green), 0.56 (D. green), 0.65 (D. green), 0.81
(L. green).
E 5 spots
0.03 (D. green), 0.05 (D. green), 0.56 (D.
green), 0.65 (D. green), 0.81 (L. green).
F 4 spots
0.03 (D. green), 0.05 (D.green), 0.56 (L.
green), 0.65 (L. green).
L - Light, D - Dark.

29. Rf values of TLC at 366nm.
TLC
analysis
Sample
No. of
Spots
Rf values
Under UV
366nm
RW 1 spot 0.58 (F.L. blue).
A 1 spot 0.58 (F.L. blue).
B 0 spots -
C 0 spots -
D 2 spots 0.55 (F.L. blue), 0.67 (F.L. blue).
E 4 spots
0.22 (F.L. blue), 0.39 (F.L. blue), 0.53 (F.D.
blue), 0.67 (FD. blue).
F 1 spot 0.22 (F.L. blue).
L - Light, D - Dark, F - Fluorescence.

30. Rf values of TLC after Post chromatographic
derivatisation.
TLC analysis Sample
No. of
Spots
Rf values
Post
chromatographi
c derivatisation
RW 5 spots
0.42 (L. pink), 0.45 (L. pink), 0.56 (D. pink),
0.65 (D. pink), 0.84 (L. purple).
A 5 spots
0.26 (L. pink), 0.48 (L. pink), 0.56 (D. pink),
0.65 (D. pink), 0.84 (L. purple).
B 3 spots 0.26 (L. pink), 0.56 (L. pink), 0.65 (L. pink).
C 3 spots 0.26 (L. pink), 0.56 (L. pink), 0.65 (L. pink).
D 5 spots
0.38 (L. pink), 0.56 (D. pink), 0.65 (D. pink),
0.79 (L. pink), 0.84 (L. pink).
E 4 spots
0.04 (D. brown), 0.06 (D. brown), 0.48 (L.
pink), 0.84 (L. purple).
F 5 spots
0.14 (L. pink), 0.27 (L. pink), 0.56 (L. pink),
0.65 (L. pink), 0.84 (L. purple).
L - Light, D - Dark.

31. TLC photo documentation of chloroform extract of
Croton tigilium raw sample and sample A, B, C, D, E, F.
254nm. 366nm.
Post Chromatographic derivatisation

32. Discussion on Physico-chemical Parameters:
Foreign matter: In Sample - RW, the foreign matter is found
to be 6.8%, the percentage of foreign matter is higher
according to API, and it should not be more than 2%. Higher
percentage of foreign matter indicates the adulteration and
inferiority of drug.
pH: pH value in the raw sample was 6.03 while in the
Shodhita sample it was varying according to the Shodhana
medium used. In samples where milk was used as Shodhana
medium i.e. in method - A,B,C there was slight variation in
the pH value, as pH of milk ranges between 6.5-6.7. In
method - D, E it was more acidic than the raw
sample, because of the lemon juice used as the medium for
Shodhana.

33. Loss on Drying: Loss on drying (at 105C) of sample - RW is
3.92% w/w, sample - A is 1.78% w/w, sample - B is 6.75%
w/w, sample - C is 1.81% w/w, sample - D is 6.02%
w/w, sample - E is 3.71% w/w and sample - F is 2.55% w/w.
Loss on drying indicates the moisture content of the drug, here
the moisture content was less in Sample A and C and more in
Sample B and D, because of various different processes.
Ash Value: Ash value depends upon the inorganic
substances present in the particular drug, this parameter has
importance in quality control and standardization of drugs.
More the inorganic substances present in drugs higher will be
the ash value. Ash value of sample - RW is 2.80% w/w, sample
- A is 2.74% w/w, sample - B is 4.96% w/w, sample - C is
2.61% w/w, sample - D is 2.37% w/w, sample - E is 2.99% w/w
and sample - F is 2.68% w/w. In Sample - B, ash value is
higher because of its Shodhana with the Tankana and more
heating process involved.

34. Acid insoluble ash: In case of sample - RW is 0.50%
w/w, sample - A is 0.48% w/w, sample - B is 0.51%
w/w, sample - C is 0.48% w/w, sample - D is 0.47%
w/w, sample - E is 0.53% w/w and sample - F is 0.44% w/w.
So, it is found within the range not more than 0.5% in many
samples. All samples had same acid insoluble ash
suggesting their same Bio- availability.
Water soluble extractive value: Various components have
their solubility in particular media. In this study, soluble
principles of the samples were examined in water and
alcohol; Water soluble extractive indicates the Bio-
availability of the drug in the water media. Results obtained
in sample - RW is 7.64% w/w, sample - A is 1.88%
w/w, sample - B is 4.44% w/w, sample - C is 1.42%
w/w, sample - D is 3.37% w/w, sample - E is 8.44% w/w and
sample - F is 1.52% w/w.

35. Alcohol soluble extractive value: These values of sample
– RW is 29.16% w/w, sample - A is 22.58% w/w, sample - B
is 23.88% w/w, sample - C is 15.37% w/w, sample - D is
24.78% w/w, sample - E is 22.64% w/w and sample - F is
16.83% w/w. The drug Jayapala is more soluble in alcohol
media.

36. Quantitative estimation of Croton oil in Raw and
Shodhita Jayapala:
The percentage of croton oil was estimated in the raw
sample and all the purified samples after complete drying of
the Jayapala endosperm.
No marked difference was found in the croton oil % in the
sample A, C, D, E and F. This may be because of the loss of
the moisture content during the drying process of the
Shodhana Samskara which may have resulted in the
increase in the concentration of the oil.
In sample B the croton oil % was 18.04% in comparison to
the raw drug 50.77%, nearly there was 32.73% decrease of
oil content in comparison to the raw drug. This may be
because of the Shodhana media employed, here Tankana
was used along with the Dugda for the Swedana Samskara.

37. HPTLC Analysis:
The chloroform extract of all the Shodhita samples and Raw
sample were used for the chromatographic study. Toluene :
Ethyl acetate: (6:1) was used as Solvent system.
TLC as well as HPTLC was carried out and the developed
plates were visualized in UV 254, 366 and then derivatised with
vanillin sulphuric acid reagent and scanned under UV 254 and
366 nm.
Total number of 6 spots were found in Raw Sample, 7 in
Sample-A, 2 in Sample-B, 3 in Sample-C, 6 in Sample-D, 5 in
Sample-E and 4 spots in Sample-F were observed in short UV.
Under long UV 1 spot was found in raw Sample, 1 in Sample-
A, 0 in Sample-B, 0 in Sample-C, 2 in Sample-D, 4 in Sample-E
and 1 spot in Sample-F and after derivatised with vanillin
sulphuric acid 5 spots were found in Raw Sample, 5 spots in
Sample-A, 3 in Sample-B, 3 in Sample-C, 5 in Sample-D, 4 in
Sample-E and 5 spots in Sample-F.

38. According to observations, commonness in the Rf values
was found in some of the the samples and also some new
spots were also found.
This is indicative of formation of some new chemical
compounds in all the purified samples utilizing different
media; it may be due to the various physical and chemical
changes taking place during the Shodhana process.
However, in absence of marker compound CROTIN, it is
impossible to draw any conclusion from the TLC and HPTLC
profile. Nevertheless, it can be considered as a preliminary
data for laying the standards for the Shodhana Samskara of
Jayapala, by utilizing different media and methods.

39. The Shodhana Samskara is not only a method which takes
away the physical and chemical impurities from the Visha
Dravya but also it potentiate the Dravya by adding useful
ingredient into it.
The macro, micro, phyto-chemical and physiochemical
results obtained in the pharmacognostical study are within
the pre-established standards in the literature for the drugs
(API).
The Pharmacognostical standards of Jayapala discussed
here can be considered as the identifying parameters to
substantiate and authenticate the drug in future.
The pharmaceutical techniques employed in these Shodhana
Samskara helps for the purification, detoxification, and
potentiating of the drug Jayapala.

40. Phytochemical analysis of the croton oil suggests that, the
Shodhana Method - B has been found better in comparison
to other methods on the basis of quantitative estimation of
croton oil.
Swedana Samskara done by utilizing Godugda and Tankana
can be considered as a better media for Shodhana of
Jayapala in comparison to Godugda alone, or with other
Shodhana Samskaras.
However, in absence of marker compound CROTIN - a
Toxalbumin, it is impossible to draw any conclusion from the
TLC and HPTLC profile. Nevertheless, it can be considered
as a preliminary data for laying the standards for the
Shodhana Samskara of Jayapala, by utilizing different media
and methods of Shodhana.