acute toxicity study - virgo uap · pdf filethis study determines acute toxicity of...

Acute Oral Toxicity Study on

SOBEREX CAPSULE

OF

Sponsor Study Centre:

VIRGO UAP PHARMA PVT.LTD. PHARMACOLOGY LABORATORY MAHAGUJARAT IND.ESTATE , SDM CENRE FOR RESEARCH IN PLOT NO. 423/98-B. AYURVEDA AND ALLIED SCIENCES SARKHEJ – BAVLA HIGHWAY LAXMI NARAYANA NAGAR, KUTHPADY, VILL – MORAIYA, TA – SANAND UDUPI - 574118 DIST – AHMEDABAD - 382213 Email: [email protected] Web: www.virgouap.com

http://www.virgouap.com/

2

STUDY REPORT

Evaluation of “Soberex Capsule” for safety

profile

Acute Oral Toxicity Study

Project Director

Dr. B. Ravishankar

Chief Investigator

Mr. Ravi M

Pharmacology Laboratory

SDM Centre for Research in Ayurveda and Allied Sciences, Kuthpady, Udupi -574118

3

INDEX

Title

Page No.

Index

3

Investigators conclusion

4

Details of experiments

5

Materials and methods

6-9

Observation

10

Result

11

Conclusion

12

Data sheets 13-22

4

Acute toxicity study:

This study determines acute toxicity of “Soberex Capsule” an oral formulation prepared

with suspension of test drug using gum acacia, in rats was assessed for single dose acute

toxicity by employing OECD guidelines 425 using AOT software. The dosed (up down as per

requirement) rats were observed for 14 days, for general appearance, cage side behaviour

including increased or decreasing motor activity, convulsions, straub’s reaction, catatonia,

muscle spasm, spasticity, ophisthotonus, hyperesthesia, muscle relaxation, anaesthesia,

arching and rolling, lacrimation, salivation, diarrheal, writhing movement, mode of respiration

and changes in skin colour etc., with mortality and autopsy finding in case of dead animal.

Conclusion:

The test drug did not produce any mortality up to the dose of 2000mg/kg per oral which is

equivalent to 22.4g total dose for a human being weighing 70 kg man. At the dose level

studied the drug also did not produce any observable toxic effect except for mild irritation in

animal in the dose550mg/kg and 2000mg/kg. Based on the observation made and recorded it

can be concluded that the test drug is without any toxic potential even at the dose of

2000mg/kg in animals equivalent to 22.4g for human being. This dose in human being is 11

fold in comparison to normal human dose 2.04g per day.

Mr. Ravi M Dr .B. Ravishankar

M.Sc (Medical Pharmacology) M.Sc (Med Pharmacology)., Ph.D

Research Officer Director and Professor or Exptl Medicine

Pharmacology and Toxicology Lab

SDM Centre for Research in Ayurveda and Allied Sciences

Udupi- 574 118

Karnataka State

5

Acute oral toxicity study of test drug “Soberex Capsule” on rats

Date of experiment : 06-01-12

Acclimatization of animals : 7 days

Date of treatment : 13-01-12

End of experiment : 05-02-12

Observation period : 14 days

Regulatory guidelines followed : OECD test guideline No. 425

IAEC approved number : IAEC-12/11-SDMCRA SP/04

6

2. Materials methods:

2.1 Details of test formulation:

1. Name of the test formulation : Soberex Capsule

Soberex Capsule : Information about name and quantity of the ingredients Ingredients name

Botanical name Part or form used

Book Ref Quantity

1. Ext. Brahmi Bacopa monnieri Whole Plant A.P.I. 50 mg

2. Ext. Jatamansi Nardostychas jatamsnsi Rhizome A.P.I. 30 mg

3. Ext. Shankhpushpi Convolvulus pluricaulis Whole Plant A.P.I. 50 mg

4. Ext. Sarpagandha Rauwolfia serpentina Root A.P.I. 10 mg

5. Vacha Acorus calamus Rhizome A.P.I. 30 mg

6. Ashwagandha Withania somnifera Root A.P.I. 20 mg

7. Arjun Terminalia arjuna Stem Bark A.P.I. 30 mg

8. Akik Bhasma ---- Powder R.T.S. -1 10 mg

9. Uste khudus Lavandula stoechas Flower N.A.VOL.-2 10 mg

10.Jyotismati Seed Celestrus paniculatus Seed A.P.I. 50 mg

11.Khurasani Ajama Hyocyamus niger Seed A.B. 50 mg

12.Excipients Q.S.

2. Batch number : SCA01( Trial batch for product permission and mfg. Licence)

3. Manufacturing date : December 2011

4. Expiry date : November 2016

5. Storage condition : Cool and dry place

6. General use : Stress reliever and memory enhancer.

7. CAS number : APPLIED

9. Country of origin : India

10. Manufacturer’s name : VIRGO UAP Pharma Pvt.Ltd.

Mahagujarat Ind.Estate Plot no. 423/ 98-B, Sarkhej - Bavala

highway, Vill - MORAIYA, Ta - Sanand Dist - AHMEDABAD -

382213(GUJARAT)

7

2.2- TEST CONDITIONS

1. Animal species : Rats

2. Strain : Wistar albino

3. Source : Animal house attached to SDM Research centre, SDM

Ayurveda College Udyavara

4. Selection : A total of 5 healthy either sex of body weight 150-200g

Rats were selected according to AOT software .

5. Acclimatization period : All the selected animals were kept under

acclimatization For 7 days before dosing.

7. Numbering and identification : The animal were marked with saturated

Picric acid solution in water for proper

identification. The marking within the

cages is as follows.

Animal number

Marking

1 Head

2 Neck

3 Middle of the back

4 Base of the tail

5 No mark

8

The group number, animal number and sex of the animal were identified with the help of cage cards,

as presented in following table.

Sl. no

Identification of animals

Desired dose (according to AOT)

Body weight (grams )

Calculated dose (ml)

1 Head 175mg/kg 220 1.13ml

2 Neck 550mg/kg 130 0.7 ml

3 Back 2000mg/kg 175 3.4ml

4 Base of the tail 2000mg/kg 190 3.7ml

5 No mark 2000mg/kg 200 3.92ml

Husbandry condition:

1. Housing : Rats were housed in each cage of poly propylene with stainless steel top grill. The dry husk was used as bedding material and was changed every morning.

2. Environment : The animal were exposed to 12 hours light and 12 hours dark cycle with The relative humidity 50 to 70 % and the ambient temperature was

22 ± 03®c.

3. Diet : Amruth brand rat pellete feed supplied by Pranav Agro Ltd, was provided

throughout the study period except on previous night of dosing i.e (overnight) fasting before dosing. The drinking water was given ad libitum Propylene bottles with stainless steel sipper tube.

9

2.4 Preparation of Test formulation for administration: 1. Test drug : Soberex Capsule 2. Vehicle : Gum acacia 3. Dose preparation : The test formulation supplied by sponsor was made in to Fine suspension in vehicle with suitable concentration All the animal were dosed constant dose volume. (1 ml/ 100g body weight) 175 mg/kg , 550mg/kg, 2000mg/kg 4. Schedule : single dose per animal a) Administration : The test formulation was administered through oral route at different dose levels to respective animal through oral feeding needle on to disposable syringe b) Dose fixation : According to the AOT Software. C) Route : oral d) Dose : 175mg/kg, 550mg/kg, 2000mg/kg test substance e) Dose volume : 1ml/100g animal

10

3. Observation:

3.1 Examination of Physical and Behavioural changes :

The animal was observed continuously for 4 hours after the dosing. The careful cage side

observation was done without disturbing the animal attention and at the end of the every

hour the animal were individually exposed to open arena for recording the behavioural

changes like increased or decreased motor activity , convulsions, straub’s reaction, muscle

spasm, catatonia, spasticity, ophisthotonus , hyperesthesia , muscle relaxation, anaesthesia,

arching and rolling, lacrimation, salivation , diarrhoea, writhing , mode of respiration, changes

in skin colour etc. exitus, CNS depression – hypo activity, passivity, relaxation, ataxia,

narcosis, etc.

3.2 Mortality:

All the animals were observed at ½, 1, 2, 3, 4, 24 h, 48 h after dosing and there after

daily once for mortality during the entire period of the study (i.e.14 days).

11

4. RESULT:

4.1 Physical and behavioural examination:

There were no physical and behavioural changes-(except mild increase in motor activity and

irritability seen in animal dosed 550mg/kg and 2000mg/kg) in all the treated animals on day

one at ½, 1,2,3,4 hours intervals after dosing and there after once daily for 14 consecutive

days. Thus the data obtained from the study on single dose administration of Soberex Capsule

oral administration up to 14 days of observation period does not result in any physical and

behavioural changes.

4.2 Mortality:

All the animals belonging to the treated group were survived throughout the 14 days

observation period after dosing.

12

Conclusion:

The test drug did not produce any mortality up to the dose of 2000mg/kg per

oral which is equivalent to 22.4g total dose for a human being weighing 70 kg

man. At the dose level studied the drug also did not produce any observable

toxic effect except for mild irritation in animal in dose550mg/kg and 2000mg/kg

and thus it could be concluded that the test drug is without any toxic potential

even at the dose of 2000mg/kg in animals equivalent to 22.4g for human being.

Dr. B. Ravishankar

13


SDM Centre for research in Ayurveda & allied sciences, Kuthpadi,

Udupi

PRO FORMA FOR RECORDING SIGNS AND SYMPTOMS DURING ACUTE TOXICITY STUDIES

Group: 175mg/kg Date: 13/01/12 Dose: 1.13ml

Animal: Rat-1(220g) Route: Oral Drug: Soberex Capsule

Experiment conducted by: Head of the lab (Director):

N: Normal

SIGNS & SYMPTOMS Basal 30 min 1h 2h 3h 4h 24h 48h

General impression N N N N N N N N Increased motor activity - - - - - - - - Convulsion: Tonic - - - - - - - -

Clonic - - - - - - - - Straubs reaction - - - - - - - - Muscle spasm - - - - - - - - Catatonia - - - - - - - - Opisthotonus - - - - - - - - Hyperaesthesia - - - - - - - - Decreased motor activity - - - - - - - - Muscle relaxation - - - - - - - - Anaesthesia - - - - - - - - Arching and rolling - - - - - - - - Lacrimation - - - - - - - - Diarrhoea - - - - - - - - Writhing - - - - - - - Salivation Viscid - - - - - - - -

Watery - - - - - - - - Respiration Stimulation - - - - - - - -

Depression - - - - - - - - Failure - - - - - - - -

Skin colour Blanching - - - - - - - - Cyanosis - - - - - - - -

Vasodilatation - - - - - - - - Grip strength(kg)-mean N N N N N N N N Visual placing response N N N N N N N N Tail pinch response N N N N N N N N Auditory response N N N N N N N N mucus membrane N N N N N N N N

Piloerection - - - - - - - -

14

Pharmacology Laboratory SDM Centre for research in Ayurveda & allied sciences, Kuthpadi, Udupi

PRO FORMA FOR RECORDING SIGNS AND SYMPTOMS DURING ACUTE TOXICITY STUDIES Group: 550mg/kg Date: 16/01/12 Dose: 0.7ml



N: Normal


General impression N N N N N N N N Increased motor activity - - - - - - - - Convulsion: Tonic - - - - - - - -







15


SDM Centre for research in Ayurveda & allied sciences, Kuthpadi, Udupi




N:Normal


General impression N active active active active active N N Increased motor activity - + + + + + - - Convulsion: Tonic - - - - - - - -







16






N: Normal


General impression N N active N N N N N Increased motor activity - - + - - - - - Convulsion: Tonic - - - - - - - -







17






N: Normal


General impression N N active Active N N N N Increased motor activity - - + + - - - - Convulsion: Tonic - - - - - - - -







18



Udupi

Drug : Soberex Capsule Date : 13/01/12

Group : 175mg/kg Route: Oral

Animal: Rat-1(220g) Study: Acute oral toxicity


Rat

No.1

Tim

e of

dru

g

ad

min

istr

ati

on

Tim

e i

nte

rval

(min

)

Exit

us

CNS Depression

ANS

CNS Stimulation

An

alg

esia

Oth

ers

Hyp

o a

ctiv

ity

Pass

ivit

y

Rel

axati

on

Ata

xia

Narc

osi

s

Pto

sis

Exop

hth

alm

o

ses

Hyp

eract

ivit

y

Irri

tab

ilit

y

Ste

reoty

py

Tre

mors

Con

vu

lsio

n

Str

au

b t

ail

Basa

l

- - - - -- - - - - - - - - - -

11.3

0

30 - - - - - - - - - - - - - - -

60 - - - - - - - - - - - - - - -

12

0

- - - - - - - - - - - -- - - -

18

0

- - - -- - - - - - - - -- -- - -

24

0

- - - - - - - - - - - - - --

24

h

- - - - - - - - - - - - - - -

48

h

- - - - - - - - - - - - - - -

19



Udupi

Pro forma for assessing Gross behaviour


Group : 550mg/kg Route : Oral



Rat

No.2

Tim

e of

dru

g

ad

min

istr

ati

on

Tim

e i

nte

rval

(min

)

Exit

us

CNS Depression

ANS

CNS Stimulation

An

alg

esia

Oth

ers

Hyp

o a

ctiv

ity

Pass

ivit

y

Rel

axati

on

Ata

xia

Narc

osi

s

Pto

sis

Exop

hth

alm

o

ses

Hyp

eract

ivit

y

Irri

tab

ilit

y

Ste

reoty

py

Tre

mors

Con

vu

lsio

n

Str

au

b t

ail

Basa

l

- - - - -- - - - - - - - - - -

11.0

0

30 - - - - - - - - - - - - - - -

60 - - - - - - - - - - - - - - -

12

0

- - - - - - - - - - - -- - - -

18

0

- - - -- - - - - - - - -- -- - -

24

0

- - - - - - - - - - - - - --

24

h

- - - - - - - - - - - - - - -

48

h

- - - - - - - - - - - - - - -

20



Udupi





Experiment conducted by: Director:

Rat

No3

Tim

e of

dru

g

ad

min

istr

ati

on

Tim

e i

nte

rva

l (m

in)

Exit

us

CNS Depression

ANS

CNS Stimulation

An

alg

esia

Oth

ers

Hyp

o a

ctiv

ity

Pass

ivit

y

Rel

axati

on

Ata

xia

Narc

osi

s

Pto

sis

Exop

hth

alm

o

ses

Hyp

eract

ivit

y

Irri

tab

ilit

y

Ste

reoty

py

Tre

mors

Con

vu

lsio

n

Str

au

b t

ail

Basa

l

- - - - -- - - - - - - - - - -

11.3

0

30 - - - - - - - - + + - - - - -

60 - - - - - - - - + + - - - - -

120 - - - - - - - - - + - -- - - -

180 - - - -- - - - - - - - -- -- - -

240 - - - - - - - - - - - - - --

24 h - - - - - - - - - - - - - - -

48 h - - - - - - - - - - - - - - -

21



Udupi

Proforma for assessing Gross behaviour





Rat

No4

Tim

e of

dru

g

ad

min

istr

ati

on

Tim

e i

nte

rval

(min

)

Exit

us

CNS Depression

ANS

CNS Stimulation

An

alg

esia

Oth

ers

Hyp

o a

ctiv

ity

Pass

ivit

y

Rel

axati

on

Ata

xia

Narc

osi

s

Pto

sis

Exop

hth

alm

o

ses

Hyp

eract

ivit

y

Irri

tab

ilit

y

Ste

reoty

py

Tre

mors

Con

vu

lsio

n

Str

au

b t

ail

Basa

l

- - - - -- - - - - - - - - - -

11.3

0

30 - - - - - - - - - + - - - - -

60 - - - - - - - - - + - - - - -

120 - - - - - - - - - + - -- - - -

180 - - - -- - - - - - - - -- -- - -

240 - - - - - - - - - - - - - --

24 h - - - - - - - - - - - - - - -

48 h - - - - - - - - - - - - - - -

22



Udupi


Drug : Soberex Capsule Date :23/01/12




Rat

No.5

Tim

e of

dru

g

ad

min

istr

ati

on

Tim

e i

nte

rval

(min

)

Exit

us

CNS Depression

ANS

CNS Stimulation

An

alg

esia

Oth

ers

Hyp

o a

ctiv

ity

Pass

ivit

y

Rel

axati

on

Ata

xia

Narc

osi

s

Pto

sis

Exop

hth

alm

o

ses

Hyp

eract

ivit

y

Irri

tab

ilit

y

Ste

reoty

py

Tre

mors

Con

vu

lsio

n

Str

au

b t

ail

Basa

l

- - - - -- - - - - - - - - - -

11.3

0

30 - - - - - - - - - + - - - - -

60 - - - - - - - - - + - - - - -

120 - - - - - - - - - + - -- - - -

180 - - - -- - - - - - - - -- -- - -

240 - - - - - - - - - - - - - --

24 h - - - - - - - - - - - - - - -

48 h - - - - - - - - - - - - - - -

acute toxicity study - virgo uap · pdf filethis study determines acute toxicity of...

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