ACUTE AND SUBACUTE TOXICITY STUDY IN GUNMATHIKKU NILAVAGAI CHOORANAM

ACUTE AND SUBACUTE TOXICITY STUDY IN GUNMATHIKKU

NILAVAGAI CHOORANAM

Jency Risha T.*1 and Priyanka2

1

Assistant Professor, Maria Siddha Medical College, Attoor.

2

Siddha Practitioner, Madurai.

ABSTRACT

“GUNMATHIKKU NILAVAGAI CHOORANAM” is a herbo–

mineral drug. The purpose of the present study is to prove that

“GUNMATHIKKU NILAVAGAI CHOORANAM” is non-toxic. If

there is no adverse reaction on animal during experimental studies then

the drug can be used for clinical practice. In this study the

herbo-mineral drug has been tested as non-toxic drug. Because the acute and

sub acute toxicity study proved non-toxic activity of the

“GUNMATHIKKU NILAVAGAI CHOORANAM”.

KEYWORDS: Gunmathikku nilavagai chooranam, acute toxicity, animal study, sub acute toxicity, siddha drug.

INTRODUCTION

Siddha medicine is a system of traditional medicine of Tamilians and they give us remedies

for both physical and mental illness. Siddha system of medicines relies on herbs, minerals,

metals and animal products. Today toxicological analysis is must for a drug to find non toxic,

according to world’s standard. While using the crude drugs for medicine preparations, they

are subjected to purification. Several purification methods are described by the siddhars for

different crude drugs.

Toxicology is the branch of medical science which deals with poisons with reference of their

source, characters and properties etc. Toxicity study is paramount in the screening of newly

developed. The Toxicity studies of Siddha drug provide scientific basis for their traditional

use and to prove that they are safe and efficacy. Most of the toxicity study is carried out by

experimental animals. Hence the present study is aimed to evaluate the acute and subacute

Volume 8, Issue 8, 575-584. Research Article ISSN 2277– 7105

Article Received on 22 April 2019,

Revised on 12 May 2019, Accepted on 02 June 2019,

DOI: 10.20959/wjpr20198-15225

*Corresponding Author Jency Risha T.

toxicity studies in “GUNMATHIKKU NILAVAGAI CHOORANAM” in Wistar Albino Rats

and it is used for all type of Ulcers(GUNMAM).

MATERIALS AND METHODS PREPARATION OF MEDICINE

In this study “GUNMATHIKKU NILAVAGAI CHOORANAM” is selected for the toxicity

study from the text book, Sarabendirarvaidhiyamuraigal (Gunma rogasigichai) Pg.no:61, Mr.

K.Vasudevasasthri, Dr.S.Venkadarajan, year of edition: 2001.

Collection of raw drugs

The raw drugs are purchased from Gopalasan Raw drugs store at Nagercoil town.

Ingredients

 NILAVAGAI

 THIPILI

 MILAGU

 KARUNSEERAGAM

 SEERAGAM

 PERUNGAYAM

 CHUKKU

 OMAM

 INDHUPPU

PURIFICATION OF RAW DRUGS NILAVAGAI

The collected drug should be grained well and purified by pittaviyal process (Baked in waper

of milk).

THIPILI

The drug will be soaked in lime juice and make it dried.

MILAGU

The drug will be soaked in sour butter milk and roast it.

KARUNSEERAGAM

SEERAGAM

The drug will be soak limestone water and allowed to dry in sunlight.

PERUNGAYAM

The perungayam has to be roasted in ghee.

CHUKKU

The outer skin of the dry ginger was peeled off and used.

OMAM

The drug will be soak limestone water and allowed to dry in sunlight.

INDUPPU

Soak the Induppu in vinegar and distilled it. Then allow it to get dried in sunlight.

METHOD OF PREPARATION

Purify all the above drugs and powder each item individually, sift and mix them well.

Route of administration

Oral.

Dose of the drug

Verugadi (1250mg).

Adjuvant

Hotwater.

ACUTE TOXICITY STUDY IN FEMALE WISTAR ALBINO RATS TO EVALUATE TOXICITY PROFILE OF GUNMATHIKKU NILAVAGAI CHOORANAM

Guidelines followed

OECD Guidelines No.423

The experimental protocol was approved by Institutional Animal Ethical Committee as per

the guidance of Committee for the Purpose of Control and Supervision of Experiments on

Animals (CPCSEA), Ministry of Environment and Forest, Government of India. Approval

Study design and Controls

1) Female Wistar Albino Rats in controlled age and body weight wereselected.

2) GUNMATHIKKU NILAVAGAI CHOORANAM was administered at 5mg/kg, 50mg/kg,

300mg/kg, 2000 mg/kg body weight water as suspension along withblank.

3) The results were recorded on the day of drug administration approximately 1st, 3rd, 4th and 24th hour in post dosing and further made in to observation upto 14 days.

EXPERIMENTAL PROCEDURE 1. Animals

A total of 15 Female Wistar Albino Rats with an approximate age of 6 weeks are purchased

from JAWAHARLAL INSTITUTE OF POSTGRADUATE MEDICAL EDUCATION AND

RESEARCH, PUDUCHERRY-6. On their arrival, a sample of animals was chosen at random

and weighed to ensure compliance with the age requested. The mean weight of Female

Wistar Albino Rats was 100 – 150g respectively. The animals were housed in metabolic

cages (55X32.7X19cm), with sawdust litter, in such a way that each cage contained a

maximum of 3 rats of the same sex. All animals underwent a period of 14 days of observation

and acclimatization between the date of arrival and the start of treatment. During the course

of this period, the animals were inspected by a veterinary surgeon to ensure that they fulfilled

the health requirements necessary for initiation of the study.

DOSES

The doses for the study were selected based on literature search and range finding study.

Following the period of fasting, the animals were weighed and then drug was administered

orally as single dose using a needle fitted onto a disposable syringe of approximate size at the

following different doses(Table-1).

Table –1.

GROUP DOSE

Group –I Control

Group – II 5mg/kg

Group – III 50mg/kg

Group – IV 300mg/kg

Group – V 2000mg/kg

The test substance was administered as single dose. After single dose administration period,

SUB-ACUTE TOXICITY STUDY IN WISTAR ALBINO RATS TO EVALUATE TOXICITY PROFILE OF GUNMATHIKKU NILAVAGAI CHOORANAM

1. Test Guideline followed

OECD 407 Method – Sub-Acute Toxic Class Method (Repeated Dose 28- Days Oral

Toxicity Study in Rodents).

2. Test substance detail

Name: GUNMATHIKKU NILAVAGAI CHOORANAM

3. Test system detail

The study was conducted on 3 male and 3 female Wistar Albino Rats. These animals were

selected because of the recommended rodent species for oral studies as per followed

guideline and availability of animals 8-12 weeks old male and female rats were selected after

physical and behavioral examination. The body weight range was fallen within ± 20% of the

mean body weight at the time of randomization and grouping. The rats were housed in

standard laboratory condition in Polypropylene cages, provided with food and water ad

libitum. The experimental protocol was approved by Institutional Animal Ethical Committee

as per the guidance of Committee for the Purpose of Control and Supervision of Experiments

on Animals (CPCSEA), Ministry of Environment and Forest, Government of India.

Doses

The doses for the study were selected based on literature search and range finding study.

Following the period of fasting, the animals were weighed and then test substance was

administered orally as single dose using a needle fitted onto a disposable syringe of

approximate size at the following different doses (Table-2).

Table – 2 Dose level.

TEST GROUP DOSE TO ANIMALS

(mg/kg body – weight/day) NUMBER OF ANIMALS

Group – I Control 6 (3male + 3female)

Group – II

Low dose of GUNMATHIKKU

NILAVAGAI CHOORANAM

(200mg/kg of body weight)

6 (3male + 3female)

Group – III

Mid dose of GUNMATHIKKU

NILAVAGAI CHOORANAM

(400mg/kg of body weight)

6 (3male + 3female)

Group – IV

High dose of GUNMATHIKKU

NILAVAGAI CHOORANAM

(600mg/kg of body weight)

The test substance was administered as single dose for 28 days and all animals were

observed.

OBSERVATION

These observations were also performed on week ends. The observations included but were

not limited to changes in skin, fur, eyes, mucous membranes and in the respiratory,

circulatory, central nervous, autonomous systems, somatomotor activity and behavior.

Clinical signs of toxicity

All the rats were observed atleast twice daily with the purpose of recording any symptoms of

ill-health, behavioral changes and clinical signs of toxicity daily for 28days.

Blood collection

Blood was collected through retro – orbital sinus from all the animals of different groups on

29th day. The blood was collected in tubes containing Heparin/EDTA as an anticoagulant. Animals were fasted over night prior to the blood collection.

LABORATORY STUDIES

During the 28th day of treatment, samples of blood were withdrawn from the orbital sinus from each group, under weak ether anesthesia after fasting for 16 hours. The blood samples

are used to evaluate Hematological parameters like RBC, WBC, Hb and DC.

RESULTS AND DISCUSSION

Observation Control 5 mg/kg 50mg/kg 300mg / kg 2000mg / kg I. Stimulation:

Hyperactivity - - - - -

Pyloerection - - - - -

Twitching - - - - -

Rigidity - - - - -

Irritability - - - - -

Jumping - - - - -

Clonic convulsions - - - - -

Tonic convulsions - - - - -

II.Depression:

Ptosis - - - - -

Sedation - - - - -

Sleep - - - - -

Loss of Traction - - - - -

Loss of pinna reflex - - - - -

Loss of Musle tone - - - - -

Analgesia - - - - -

III.Autonomic effect:

Straub tail - - - - -

Laboured respiration - - - - -

Cyanosis - - - - -

Reddening - - - - -

Abnormal secretions - - - - -

IV. Mortality:

After 24 hours - - - - -

GUMATHIKKU NILAVAGAI CHOORANAM was administered single time at the dose of

5mg/kg, 50mg/kg, 300mg/kg and 2000mg/kg to rats and observed for consecutive 14 days

after administration. Doses were selected based on the pilot study and literature review. All

animals were observed daily once for any abnormal clinical signs. Weekly body weight and

food consumption were recorded. No mortality was observed during the entire period of the

study. Data obtained in this study indicated no significance physical and behavioral signs of

any toxicity due to administration of GUMATHIKKU NILAVAGAI CHOORANAM at the

doses of 5mg/kg, 50mg/kg, 300mg/kg and 2000mg/kg to rat.

At the 14th day, all animals were observed for functional and general behavioral examination. In functional and general behavioral examination, home cage activity, hand held activities are

normal. Functional and general behavioral examination was normal in all treated groups.

From acute toxicity study it was observed that the administration of GUMATHIKKU

NILAVAGAI CHOORANAM up to the dose of 2000mg/kg to the rats do not produce

drug-related toxicity and mortality. So No-Observed-Adverse- Effect-Level (NOAEL) of

SUB-ACUTE TOXICITY RESULTS

Following figures shows the result of heamotological profiles

Fig: 1.

Fig: 2.

Fig: 4.

Fig: 5.

Fig: 7.

CONCLUSION

From acute toxicity study shows that GUMATHIKKU NILAVAGAI CHOORANAM did not

produce any toxic effect and mortality at dose of 5mg/kg, 50mg/kg, 300mg/kg and

2000mg/kg to rats. So NO-Observed-Adverse-Effect-Level (NOAEL) of GUMATHIKKU

NILAVAGAI CHOORANAM is 2000mg/kg.

From sub acute toxicity study shows that GUMATHIKKU NILAVAGAI CHOORANAM can

be considered safe, as it did not cause either any lethality or adverse changes with general

behavior of rats and also there were no observable detrimental effects (200 to 600mg/kg body

weight) over a period of 28days. Our results have demonstrated that the GUMATHIKKU

NILAVAGAI CHOORANAM is relatively safe when administered orally for all type of

ulcers (GUNMAM) in human for long period.

REFERENCES

1. T.V.Sambasivam Pillai, Dictionary of Medicine, Chemistry, Botany and allied science

vol-II, published by Research Institute of Siddhar’s Science, Chennai, 1931, First Edition.

2. Marunthu Sei Iyyalum Kalaium (Page No: 283,284,285) Deva Asirvatham Samuel MD(s)

3. Dr.K.N.Kuppusamy Mudaliyar-HPIM.

4. Dr.K.S.Uthamarayan, Siddha Vaidhiya Thiratu.

5. Supplement to glossary of India Medicinalplants.