From,

DR.JAGADISH NAYAKA M.R

I P.G Scholar

Department of Post Graduate Studies in Rasashastra

J S S AyurvedaMedicalCollege

Mysore.

To,

The Registrar

RajivGandhiUniversity of Health Sciences,

Bengaluru,

Karnataka.

Through,

The Principal and Head of the Department

Of P G studies in Rasashastra.

J S S AyurvedaMedicalCollege.

Mysore .

Respected Sir,

Sub: Submission of completed proforma for Registration of subject for dissertation.

I request you to kindly register the below mentioned subject against my name for the submission of dissertation to the Rajiv Gandhi University of Health Sciences, Bengaluru, for the partial fulfillment of M.D (Ayurveda) in Rasashastra.

Title of the dissertation:-

“PHARMACEUTICO-ANALYTICAL AND COMPARATIVE TOXICITY STUDY OF SHUDDHA HARATALA AND HARATALA BHASMA ”

Here with I am enclosing completed proforma for registration of the subject for dissertation.

Thanking you,

Yours faithfully

DATE:

PLACE:

DR.JAGADISH NAYAKA M.R

Rajiv Gandhi University of Health Sciences; Karnataka; Bengaluru.

ANNEXURE 2

Proforma for registration of subject for Dissertation:

1. / Name of the candidate
Address / Dr.JAGADISH NAYAKA, B.A.M.S
I.P.G scholar
Department of Post Graduate studies in Rasashastra.
JSS.Ayurveda medical College.
Mysore
Residential address:
Dr. JAGADISH NAYAKA
#762 4TH Main 8th cross T.K.Layout
Mysore
2. / Name of the institution / JSS.AyurvedaMedicalCollege
Mysore
3. / Course of the study & subject / Ayurveda vachaspathi
M.D (Ayurveda) in Rasashastra
4. / Date of admission to course / 13-09-2010
5. / Title of the topic / “PHARMACEUTICO-ANALYTICAL AND COMPARATIVE TOXICITY STUDY OF SHUDDHA HARATALA AND HARATALA BHASMA ”

6. Brief resume of intended work:-

6.1 Need for the study

Haratala is one of the Uparasas, mentioned in Ayurveda texts1. Haratala is known for itstherapeutic efficacy in Phiranga, Vatarakta, Visarpa, Vicharchika Vataroga, Sleshmaroga, Rakthapitta, Kusta, Kandu, Dadru, Pama, Slipada, Visharoga, Vishamajwara, Prameha, Apasmara, Nadivrana, Arshas, Kasa, Swasa, Vrana, Rakthavikara, Kaphapittaja Roga2.

It is widely used in formulations like :-

RASA MANIKYA

SAMIRAPANNAGA RASA

SMRUTHISAGARA RASA

TALAKESHWAR RASA

TALA SINDURA

NITHYANANADA RASA

KASTHURI BHAIRAVA RASA etc,

Inspite of having wide therapeutic usage of Shuddha Haratala & Haratala Bhasma, there is a fear of Arsenic toxicity in global scenario.

Haratala is known to cause toxic effects like Daha, Kshobha, Toda, Kampa, Raktha Vikara, Kusta , and Vata Kaphaja Rogas finally it may lead to death also, when administered either without Shodhana or improper Shodhana and improper administration of Shodhita Haratala ,3,4.

Hence this study is intended to evaluate the toxicity and ensure safety of Shuddha Haratala and Haratala Bhasma, by means of animal experimentation.

6.2Review of literature.

References regarding Haratala as one of the Uparasas is seen in Rasaratna Samuchaya 1 and in many other Rasashastra texts .

Haratala is of 2 types- Patra Haratala and Pinda Haratala. Patra Haratala is golden in colour, heavy and snigdha. It contains small layers and is bright, which is more potent 3 .

Haratala Shodhana and Marana has mentioned in Rasaratna Samuchya3 and other Rasashastra texts.

Shuddha Haratala and Haratala Bhasma gunas has mentioned in Rasendra Sara Sangraha5, Rasatarangini6

Specific Haratala Bhasma pareeksha and dosage is mentioned in Rasatarangini6.

Toxic effects of improper Shodhana or improper administration of Haratala is mentioned in Rasaratna Samuchya3, Rasatarangini4 and other Rasashastra books.

Previous works published

  1. Haratala Bhasma nirmana evam raktha kano par adhyana by Dr.Krishna,1990,Patiala.
  2. A standard preparation of Haratala Bhasma and its therapeutical efficacy in certain skin disorder by Dr. Pillai,1991,Jamnagar.
  3. Haratala Basma ka vividha vidhiyo dwara nirmanathmaka evam kusta roga par prabhavatmaka adhyana by Dr. Kaushik,1993,Jaipur.
  4. Analytical study of Sodhitha Haratala and Haratala Bhasma and assessment of their Spermatogenic activity on Albino rats. Dr .Sasidharan.J, 2003, Mallapura.
  5. Pharmaceutico analytical study of Haratala Bhasma and its anti-convulsant effect on Abino rats. Dr Ajith Narayan, 2004, Bellary.

6.3Aims & objectives of study.

  1. To carry out Shodhana and Marana of Haratala as per the classical reference.
  2. To carry out physico-chemical analysis of Shuddha Haratala and Haratala Bhasma.
  3. To carry out comparative toxicity study of Shuddha Haratala and Haratala Bhasma in Wister albino rats (females).

7. Materials & Methods

7.1 Source of Data:-

Literary data:- will be collected from authentic sources like Classical books, Articles , Publications, Journals & Internet.

Raw material data:-Genuine samples of Haratala and the drugs required for Shodhana and Marana will be collected from the market.

Source of Animal: - From animal house of JSS Pharmacy college, Mysore.

7.2 Method of collection of data,

Grahya lakshanayukta Haratala will be collected 3.

Shodhana of Haratala will be done by Swedana with Kushmanda swarasa in Dola yantra for 3 hours and washing with water 3.

Bhasma preparation: Shuddha Haratala will be given 3 bhavanas with thick decoction of Palasha moola, chakrikas will be made, placed inside sharava samputa and putas of 10 cow dung cakes will be given and such 12 putas are necessary to obtain Bhasma 3 .

Shuddha Haratala & Haratala Bhasma will be subjected to Physico- chemical analysis like

1. Organoleptic tests.

2. Bhasma pareekshas including Nirdhuma pareeksha.

3. N.P.S.T.

4. Atomic Absorption Spectrometry

5. Scanning Electron Microscopy.

Toxicity of Shuddha Haratala & Haratala Bhasma will be studied through animal experimentation (Wister albino rats).

Experimental Study Design:-

Albino rats (females) weighing about 150gms to 200gms will be selected and maintained in standard laboratory condition along with proper supply of rat feed and potable water in sufficient quantity. After procuring experimental animals it will be allowed to acclimatize to a new atmospheric condition for 7 days,

Pilot study to ascertain the proper toxic doses will be conducted and depending on results grouping of animals will be done and LD50 will be ascertain7.

Study will be conducted in 2 phase

1. Acute toxicity study (1dose/24hours observation for 14 days)7

2.Sub-acute toxicity study (repeated doses for 28 days)7

In every study 6 experimental animals will be used in each groups and depending upon the pilot study results number of groups will be fixed and will be carried out for Shoditha, Marita Haratala and will be administered at different dosages with suitable media decided only after pilot study.

Replacement of animals (inclusion & exclusionCriteria) will be carried out as per OECD guidelines.

Inclusion criteria :-

Healthy Albino rat weighing between 150-200 gms will be taken.

Exclusion criteria: -

Specimen not fulfilling the above conditions, those which are ill, those which are pregnant.

Toxicity study:-

Acute and sub-acute toxicity study will be carried out as per OECD guidelines7.

Observation:-

Changes pertaining to behaviour, food and water intake, body weight will be noted every day.

Animal which will die in between the study course will be immediately autopsied and vital organs will be collected and preserved for Histo-pathological studies.

Experimental animals will be sacrificed after 28 days and autopsy will be done to observe changes in internal organs and also samples will be collected for Histo-pathological study.

Slides will be prepared by the authentic methods and visualized under microscope to see Histo-pathological changes and will be compared with the control group animals.

7.3 Does the study require in investigations of interventions to be conducted on patients or other human or animals? If so please describe briefly.

Yes, Histo-pathological changes will be investigated

7.3 Has ethical clearance been obtained from your institution in case of 7.3?

Yet to be obtained.

8. Lists of reference.

  1. Satpute Ashok.D, Rasaratna Samuchchaya , Chaukhamba Sanskrit Pratishthan ,Varanasi,

First Edition 2003, p-53.

  1. Sarma Sadananda, Rasatarangini, Mothilal Banarasidas, Delhi,2009,edition, p-242.
  2. Satpute Ashok.D, Rasaratna Samuchchaya , Chaukhamba Sanskrit Pratishthan ,Varanasi,

First Edition 2003, p-66,67.

  1. Sarma Sadananda, Rasatarangini, Mothilal Banarasidas, Delhi,2009,edition, p-246.
  2. Satpute Ahok.D, Gopala krishna, Rasendra sara sangraha, Chaukhamba krishanadasa

academy ,Varanasi, First Edition 2003, p-100

  1. Sarma Sadananda, Rasatarangini, Mothilal Banarasidas, Delhi,2009,edition, p-252-253
  2. OECD Guideline for Acute and Sub-acute toxicity study.