SAFETY EVALUATION OF SIDDHA FORMULATION AKIL KATTAI

www.wjpps.com Vol 6, Issue 10, 2017.

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Sathiya et al. World Journal of Pharmacy and Pharmaceutical Sciences

SAFETY EVALUATION OF SIDDHA FORMULATION AKIL KATTAI

CHOORANAM BY ACUTE AND SUB-ACUTE TOXICITY STUDIES IN

WISTAR RATS

X. Helen Sathiya*1, N. Anbu

2, P. Parthibhan

3 and K. Kanakavalli

4

1P.G. Scholar, Post Graduate Department of Maruthuvam, Government Siddha Medical

College, Arumbakkam, Chennai 600 106, Tamil Nadu, India.

2Professor, HOD, Post Graduate Dept. of Maruthuvam, Government Siddha Medical College,

Arumbakkam, Chennai 600 106, Tamil Nadu, India.

3Joint Director, Indian Medicine and Homeopathy, Chennai 600 106, Tamil Nadu, India.

4Principal, Government Siddha Medical College, Arumbakkam, Chennai 600 106, Tamil

Nadu, India.

ABSTRACT

According to the recent regulatory guidelines preclinical toxicity

evaluation of the siddha formulations is mandatory to ascertain the

possibility of adverse event in humans upon short and long term usage

of the drugs. The main aim of the present research work is to evaluate

the safety of the traditional siddha formulation Akil Kattai Chooranam

(AKC) and to establish the toxicity profiling by acute and sub-acute

toxicity studies in accordance with OECD guidelines. In short term

acute toxicity study the drug AKC was administered in single doss of

2000 / kg b.w (p.o). Potential drug toxicity related to C.N.S, A.N.S and

C.V.S were observed up to 14 days. In sub-acute toxicity study the

drug AKC was administered at two dose level such as low and dose of

200 and 400 mg / kg b.w (p.o) for four weeks, Results obtained from

the acute and sub-acute study reveals that the drug AKC doesn’t reveal any significant

change in body weight, behavior and mortality in treated rats. Throughout the study period no

sign of toxicity was registered. Further it was observed that AKC at both the dose level did

not modify the weight index, food and water intake in treated animals. There is no significant

change in hematological, biochemical and histopathological observation of animals treated

with AKC at both the dose level of 200 and 400 mg / kg b.w (p.o) when compare to control

*Corresponding Author

Dr. X. Helen Sathiya

P.G.Scholar, Post Graduate

Department of Maruthuvam,

Government Siddha Medical

College, Arumbakkam,

Chennai 600 106, Tamil

Nadu, India.

Article Received on

15 August 2017,

Revised on 04 Sept. 2017,

Accepted on 25 Sept. 2017

DOI: 10.20959/wjpps201710-10295

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 6.647

Volume 6, Issue 10, 1202-1225 Research Article ISSN 2278 – 4357


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group animals. From the result it was concluded that the siddha preparation AKC offers wide

margin of safety in tested rodents and further long term usage of drugs will be considered as

safe for the ailment of various disease.

KEYWORD: Siddha, Akil Kattai Chooranam, OECD, Toxicity profiling, Acute study, Sub-

acute.

1. INTRODUCTION

Toxicity profiling of siddha preparations are become highly essential in order to prove the

safety of the formulation upon short and long term administration in humans. Results of

toxicity study render some useful information to the investigators with respect to the effect of

the drug on CNS, CVS, ANS and other metabolic organs. It provides a base for fixation of

dose to the pharmacological study. Toxicity study further reveals the information about LD50

and also the dose which causes lethal effect and also safe therapeutic dose.

The OECD Guidelines for the testing of chemicals are a collection of the most relevant

internationally agreed testing methods used by government, industry, and independent

laboratories to characterize potential hazards of new and existing chemical substances and

chemical preparations, mixtures. They cover tests for the physico-chemical properties of

chemicals, human health effects, environmental effects, and degradation and accumulation in

the environment.[1]

Animals have been used as models for centuries to predict what chemicals and environmental

factors would do to humans.[2]

Siddha preparation which is not purified properly and also

preparations with contaminants like pesticides, organic chemicals, and heavy metals exert

their toxic effect on the hematopoietic system and may render anemia, neutropenia, leukemia,

thrombocytopenia, mutagenesis, apoptosis, myelotoxicity, anemia, immunomodulation etc.

Exposure to benzene and its metabolites leads to myelodysplastic syndromes, leukemia,

lymphomas and bone marrow aplasia. Mercury and chrome affect the immune system by

immunosuppression and by evoking autoimmune reactions. Dithiocarbamates are suspected

to induce leukemia. An analysis of the pathophysiology of individual substances reveal

universal toxic mechanisms.[3]

Liver a major organ for drug metabolism predominantly exposed to the adverse effect of the

toxic chemicals. Wide of range of toxicants disturbs the liver function includes insecticides,


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metabolites, heavy metals, pesticides etc. Hepatotoxicity nature of the drugs characterized by

changes in liver function test ascertained by estimation of SGOPT and SGPT level. Liver of

limited capacity of self-healing/ regeneration but chronic administration of certain chemicals

cause abnormal liver function which in turn hinders the physiology of metabolism. Toxic

nature of compounds causes’ pathological damage in the liver was manifested by sinusoidal

congestion (SC), RBC deposition in the vein and inflammatory response. Although drug

biotransformation generally parallels a detoxication process, the bioactivation is frequently

the major cause of hepatocyte injury.[4]

With regard to the bioactivation of drugs, attention

has been mostly paid to Phase I metabolites of drugs.

Renal toxicity of the drug will be evaluated by serum creatinine and blood urea nitrogen

level. According to the literature there is a remarkable nephrotoxicity caused by specific

medicinal herbs.[5]

Adverse unadulterated herb reactions resulting in notable kidney

manifestations are usually caused by an abuse of the substance or ignorance regarding the

herb's intended administration. The most common example involves ephedra (Ephedra sinica

spp.), which contains ephedrine.

The main aim of the present investigation is to evaluate the safety of the siddha formulation

Akil Kattai Chooranam in rodents at fixed dose level by acute and sub-acute toxicity studies

in accordance with OECD guidelines.

2. MATERIALS AND METHODS

The formulation AKC comprises of the following herbs as its ingredients.

2.1. Ingredients

1. Chandanam (Santalum album) -17.5g.

2. Agil kattai (Aquilaria agallocha Roxb) -17.5g.

3. Elam (Elettaria cardamomm) -17.5g.

4. Lavangappattai (Cinnamomum verum) -17.5g.

5. Kirambu (Syzygium aromaticum) -17.5g.

6. Sombu (Pimpinella anisum) -17.5g.

7. Ati-maduram (Glycyrrhiza glabra) -17.5g.

8. Karboki vitthu (Psoralea corylifolia) -17.5g.

9. Vetpalai arisi (Wrightia –tinctoria) -17.5g.

10. Thettran vithai (Strychnos potatorum) -17.5g.


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11. Arugam vear (Cynodon dactylon) - 17.5g.

12. Chitramutti (Pavonia Zeylanica) -17.5g.

13. Karkandu – Quantity Sufficient

2.2. Source of raw drugs

The required raw drugs are procured from a well reputed indigenous drug shop. The raw

drugs taken for study will be authenticated by the Botanist of Medicinal botany department,

Govt. Siddha Medical College, Chennai.

2.3. Preparation[6]

Take each ingredient about 17.5 gm. and made it to dry in the sun light, after that it was

ground and powdered. Then equal Quantity of karkandu was added and bottled up.

2.4. Drug Storage

The trial drug is stored in clean dry air tight container and it is dispensed to the patients in air

tight bottle.

Vehicle : Butter.

Dose : 1gm, twice a day.

Duration : 48 Days.

2.5. Toxicological Profiling of Akil Kattai Chooranam

2.5.1. Animal

Healthy adult Wistar albino rat weighing between 170-200 g were used for the study. The

animals were housed in poly propylene cages and were kept in well ventilated with 100%

fresh air by air handling unit (AHU). A 12 light / dark cycle were maintained. Room

temperature was maintained between 22 + 20C

and relative humidity 50–65%. They were

provided with food (Sai feeds, Bangalore, India) and water ad libitum. All the animals were

acclimatized to the laboratory for 7 days prior to the start of the study. The experimental

protocol was approved by The Institutional Animal Ethics Committee of Sathyabama

University, Chennai, Tamil Nadu, India. SU/CLATR/IAEC/IV/016/2016.

2.5.2. Acute toxicity Study

The animals were fasted overnight (12- 16 hrs) with free access to water. The study was

conducted with single oral administration of study drug Akil Kattai Chooranam (AKC)


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2000mg/kg (p.o). The animals were observed continuously for first 72 h and then 14 days for

emerging signs of behavioral changes, body weight changes and for mortality.

Occurrence of toxicity in animals were observed continuously for the first 4 to 24 h and

observed periodically for the next 14 days. Observation includes the change in skin, fur, eyes

and mucus membrane. Appearance of C.N.S,C.V.S and A.N.S related toxicity such as

tremors, convulsions, sedation, steric behavior, respiratory distress, cardiovascular collapse,

response to sensory stimuli, salivation, diarrhea, lethargy, sleep, coma and mortality were

observed with special attention.[7]

Body weight was recorded periodically. At the end of the experiment all animals were

subjected for gross necropsy and observed for pathological changes.

2.5.3. Sub-Acute toxicity Study

The animals were randomly divided into control group and drug treated groups for two

different doses viz. low dose (200 mg/kg b.w) and high dose (400 mg/kg b.w). The animals

were administrated with the study drug once daily for 28 days. The animals in group I

(control group) received normal saline 5 ml/kg b.w. The animals in group II received low

dose of Akil Kattai Chooranam 200 mg/kg b.w (p.o) and group III received high dose of Akil

Kattai Chooranam 400 mg/kg b.w (p.o).

The rats were weighed periodically and observed for signs of toxicity pertains to C.N.S,

C.V.S, A.N.S including behavioral changes, food - water intake and morphological changes.

At the end of 28th

day, the animals were fasted for overnight with free access to water. On

29th

day the animals were sacrificed with excess anesthesia. Blood samples were collected

from aorta and stored in EDTA (ethylenediamine –tetra actate) for Hematological analysis

and for serum generation for biochemical analysis. The vital organs including heart, brain,

lungs, spleen, kidneys, liver, stomach, testes, and ovary were harvested and carefully

examined for gross lesions. The organs were preserved in 10% formalin for histopathological

assessment and interpretation.[8]

2.5.4. Hematological analysis

Blood samples were analyzed using established procedures and automated Bayer Hematology

analyzer. Parameters evaluated include Packed Cell Volume (PCV), Red Blood Cells (RBC)

count, White blood cell count (WBC), Platelet Count, Hemoglobin (Hb), Mean cell


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Haemoglobin Concentration (MCHC), Mean Red Cell Volume (MCV), Mean Cell

Hemoglobin (MCH), Mean platelet volume (MPV), Neutrophils, Eosinophil’s, Basophils,

Lymphocytes and Monocytes.

2.5.5. Biochemical analysis[9]

Serum samples were analyzed for High Density Lipoprotein (HDL), Low density Lipoprotein

(LDL), Very low density Lipoprotein (VLDL), Triglycerides (TGL), Total Cholesterol,

Blood urea nitrogen (BUN), Creatinine, Albumin, Total Protein, Glucose, Uric acid,

Aspartate Transaminase (AST), Alanine amino Transaminase (ALT) and Alkaline

Phosphatase (ALP) using Mind ray auto analyzer model BS 120.

2.5.6. Histopathological evaluation[10]

Organs included of heart, brain, lungs, spleen, kidneys, liver, stomach, testes and ovary.

Histological slides of organs were made and observed under the microscope. The

pathological observations of cross section of these organs were performed on gross and

microscopic bases. Histological examinations were performed on the preserved tissues with

particular emphasis on those which showed gross pathological changes.

2.5.7. Statistical analysis[11]

The statistical analysis will be carried by one way ANOVA (GRAPH PAD PRISM 5

computer program). Results were expressed as mean ± standard error. A statistical

comparison was carried out using the Dunnet’s test for the control and treatment group. P-

values less than 0.05 were set as the level of significance.

3. RESULTS

3.1. Effect of AKC on clinical signs of Female rats in Acute Oral Toxicity Study

The dose of AKC used for acute toxicity study is 200mg/kg is about 2 folds higher than the

normal therapeutic dose. No mortality observed at this dose level, further no significant

change with respect to clinical signs on acute toxicity observed for (24-48 h) and a long

period (14 days). The results were tabulated in Table 1.

3.2. Effect of AKC on Body weight of Female rats in acute toxicity study

No significant change was observed in body weight of female rats treated with AKC at the

dose of 2000mg/ kg. The results were tabulated in Table 2.


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3.3. Effect of AKC on clinical signs of male and female rats in Sub-acute oral toxicity

study.

No significant toxicity was observed in rats during the 28 consecutive days of treatment via

oral route with AKC at low and high dose of 200 and 400 mg/ kg b.w. The results were

tabulated in Table 3.

3.4. Effect of AKC on Body weight rats in Sub-acute oral toxicity study

No significant toxicity was observed in rats during the 28 consecutive days of treatment via

oral route with AKC at low and high dose of 200 and 400 mg/ kg b.w. The results were

tabulated in Table 4.

3.5. Effect of AKC on food and water intake of rats in Sub-acute oral toxicity study

No significant change was observed in body weight of both male and female rats treated with

AKC at the dose of 200 and 400mg / kg b.w. The results were tabulated in Table 5.

3.6. Effect of AKC on Hematological and Biochemical parameters of male and female

rats in Sub-acute oral toxicity study

No statistically significant differences were recorded in hematological and biochemical

parameters of rats treated with AKC at the dose of 200 and 400mg / kg b.w. The results were

tabulated in Table 6-9.

3.7. Effect of AKC on Histopathological changes of male and female rat in Sub-acute

oral toxicity study

No abnormality were detected in the histopathological analysis of organs (Kidney, Heart,

Liver, Brain, Lung, Spleen and Stomach) retrieved from the rats treated with low and high

dose of AKC.

Histology of brain reveals regular marginal alignment on the neurons with promising

histology. Neurons is very intact and there were no signs of edema or degeneration were

observed in sample belongs to group I,II and III. As shown in figure 01 and 02.Microscopic

observation of heart projects prominent nucleus with regular arrangement of fibres. No

evidence of pyknotic nucleus were observed in samples belongs to group I, II and III. As

shown in figure 03 and 04.

Appearance of proximal and distal convolutes tubules of kidney was normal with no evidence

of atrophy. Lumen of distal convolutes tubule and collecting duct was normal in sample


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belongs to group I,II and III. As shown in figure 05 and 06.In liver centrilobular zone appears

normal with stable network of hepatocytes. Rare appearance of Kupffer cells with no

evidence of phagocytosis in intracytoplasmic region were observed in sample belongs to

group I, II and III. As shown in figure 07 and 08.

Histopathology of lung shows no signs of airway secretion and bronchial secretion. Bronchial

blood vessels and connective tissue appears normal with no signs of pulmonary edema. In the

stomach Intracytoplasmic zone of mucosa appears normal. As shown in figure 09 and

10.Histology of spleen shown appearance of LF – lymphoid follicle; PALS – periarterial

lymphoid sheath was normal with no significant signs of enlargement were observed in

sample belongs to group I, II and III. As shown in figure 11 and 12.

Light microscopic observation stomach reveals normal histology of gastric wall composed of

normal mucosa, muscularismucosa, submucosa, muscularispropiria and adventitia. No signs

of ulceration were observed in sample belongs to group I, II and III. As shown in figure 13

and 14. In testes presence of mature somatic cells project the perfect histomorphology of

testicular cells in this group. Primary spermatocytes with large centered nucleus and dense

chromatin were observed in sample belongs to group I,II and III. As shown in figure 15.

Uterus of female rats shown normal appearance of endometrium, myometrium and uterine

glands. Arrangement of stratum basale, functionale and surface epithelium seems normal in

samples belongs to group I,II and III. As shown in figure 16. Histopathological analysis of

ovary showing normal corpus luteum (CL) and Primordial follicles with few mature ovarian

follicles with no signs of abnormality. Appearance of antral follicle, primary oocyte and

secondary follicles are normal in sample belong to group I,II and III. As shown in figure 17.

Table 1: Effect of Akil Kattai Chooranam on clinical signs of Female rats in Acute Oral

Toxicity Study.

Parameter Group I

Clinical Signs Parameters for the duration of 14 days Test Drug 2000mg/ Kg

Number of animals observed 6 Female

Lacrimation Absence

Salivation Absence

Animal appearance Normal

Tonic Movement Absence

Clonic Movement Absence

Laxative action Absence

Touch Response Normal


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Response to Sound Normal Response

Response to Light Normal Response

Mobility Normal Response

Respiratory Distress Nil

Skin Color Normal

Stereotype behavior Absence

Piloerection Absence

Limb Paralysis Absence

Posture Normal

Open field behavior Normal

Gait Balancing Normal

Freezing Behaviour Absent

Sings of Stress and Anxiety None Observed

Muscular coordination Normal

Muscle grip Normal

Sedation Absence

Social Behavior Normal

Urine Analysis No Abnormality

Urine Colour Yellowish

Urine pH 7

Urine -Glucose Absence

Urine -Ketones Absence

Urine- Bilirubin Absence

Urine-Blood Cells Negative

Urine - Pus cells Negative

Mortality Nil

Table 2: Effect of Akil Kattai Chooranam on Body weight of Female rats in acute toxicity

study.

Group Before Treatment Weight in Gms After Treatment Weight in Gms

Group I 181.8 ± 6.94 184.5 ± 7.86

Values are mean ± S.D (n = 6 per group). Statistically using one way ANOVA followed by

Dunnett’s test.

Table 3: Effect of Akil Kattai Chooranam on clinical signs of male and female rats in

Sub-acute oral toxicity study.

Parameter Group I Group II Group III

Clinical Signs Parameters for

28 days Control

Test Drug

200mg/ Kg

Test Drug

400mg/ Kg

Number of animals observed 3 Male and 3

Female

3 Male and 3

Female

3 Male and 3

Female

Lacrimation Absence Absence Absence

Salivation Absence Absence Absence

Animal appearance Normal Normal Normal

Tonic Movement Absence Absence Absence

Clonic Movement Absence Absence Absence


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Absence Absence Absence Absence

Touch Response Normal Normal Normal

Response to Sound Normal

Response Normal Response Normal Response

Response to Light Normal

Response Normal Response Normal Response

Mobility Normal Normal Normal

Respiratory Distress Nil Nil Nil

Skin Color Normal Normal Normal

Stereotype behavior Absence Absence Absence

Piloerection Absence Absence Absence

Limb Paralysis Absence Absence Absence

Posture Normal Normal Normal

Open field behavior Normal Normal Normal

Gait Balancing Normal Normal Normal

Freezing Behaviour Absent Absent Absent

Sings of Stress / Anxiety None Observed None Observed None Observed

Muscular coordination Normal Normal Normal

Muscle grip Normal Normal Normal

Sedation Absence Absence Absence

Social Behavior Normal Normal Normal

Urine Analysis No Abnormality No Abnormality No Abnormality

Urine Colour Yellowish Pale Yellow Pale Yellow

Urine pH 6 6 6

Urine Glucose Absence Absence Absence

Urine Ketones Absence Absence Absence

Urine Bilirubin Absence Absence Absence

Urine-Blood Cells Negative Negative Negative

Urine - Pus cells Negative Negative Negative

Mortality Nil Nil Nil

Table 4: Effect of Akil Kattai Chooranam on Body weight of rats in Sub-acute oral

toxicity study.

Group Before Treatment Weight in Gms After Treatment Weight in Gms

Group I 187.3 ± 5.64 198.7 ± 6.40

Group II

Group III

Values are mean ± S.D (n = 6 per group of which 3 males and 3 females). Control and

treatment groups were compared statistically using one way ANOVA followed by Dunnett’s

test.


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Table 5: Quantitative data on the food and water intake of rats treated with Akil Kattai

Chooranam for 28 days in Sub-acute toxicity study.

Group Food intake Water intake

Group I 16.08 ± 2.47 20.42 ± 5.984

Group II 17.5 ± 1.139 35.17 ± 0.6383

Group III 16.92 ± 3.957 40.17 ± 2.046



test.

Table 6: Effect of Akil Kattai Chooranam on Haematology profile of rats in sub-acute

toxicity study.

Group WBC count

(×103 µl)

RBC

(×10 6 µl)

PLT

(×10 3 µl)

MCV

(fl)

MCH

(pg)

MCHC

(g/dl)

HGB

(g/dl)

Group I 7.983 ± 2.40 5.683±0.93 506.2±311.2 56.35±4.61 17.98±2.77 31.78±2.47 12.04±2.25

Group II 11.38±2.32 6.383±1.33 640.5±212 60.23±8.77 20.62±2.32 34.15±2.89 12.6±2.25

Group III 11.27± 2.99 6.817±1.22 696.8±328.5 60.25±8.29 19.82±2.58 30.78±1.26 12.28±2.30



test.

Table 7: Effect of Akil Kattai Chooranam on Haematology profile of rats in sub-acute

toxicity study.

Group Lymph

(%)

Mon

(%)

Neutrophils

(X 103/mm

3)

Eosinophils

(%)

Basophils

(%) MPV (fl)

Group I 75.23±10.23 2.4±1.25 2.1±0.4 1.217±0.30 0.33±0.51 5.2±1.66

Group II 72.92±3.71 3.267±0.37 2.2±0.29 1.4±0.06 0.33±0.21 5.383±0.49

Group III 84.77±6.94 3.65±1.66 2.55±1.24 1.55±0.33 0.16±0.40 5.533±0.81



test.


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Table 8: Effect of Akil Kattai Chooranam on Serum Bio-chemistry profile of rats in sub-acute toxicity study.

Group

Blood

sugar ®

(mg/dl)

BUN

(mg/dl)

Serum

creatinine

(mg/dl)

Serum total

cholesterol

(mg/dl)

Serum

triglycerides

level (mg/dl)

Serum HDL

cholesterol

(mg/dl)

Serum LDL

cholesterol

(mg/dl)

Serum VLDL

cholesterol (mg/dl)

Group I 67.17±7.16 14.33±4.45 0.43±0.08 110.3±22.6 79.5±16.2 49.17±7.96 35.5±17.4 12.98±3.27

Group II 80.17±13.2 15.67±4.96 0.6±0.22 128.8±9.13 88.67±14.4 60±23.3 35.83±14.4 17.58±2.89

Group III 80.5±18.22 9.167±2.13 0.65±0.18 105.8±15.7 76.33±7.5 65±15.5 33.5±13.5 14.45±2.52

Values are mean ± S.D (n = 6 per group of which 3 males and 3 females). Control and treatment groups were compared statistically using one

way ANOVA followed by Dunnett’s test.

Table 9: Effect of Akil Kattai Chooranam on Serum Bio-chemistry profile of rats in sub-acute toxicity study

Group Serum total protein (g/dl) Serum albumin (g/dl) (AST) (IU/ml) (ALT) (IU/L) (ALP) (IU/L)

Group I 3.633±0.96 2.917±0.53 131.2±8.25 31.83±9.02 155.3±74.8

Group II 4.167±0.81 3.45±1.0 130.7±10.7 34.33±9.4 168.2±33.5

Group III 4.45±0.63 2.46±0.59 104.8±21.2 24.17±6.30 145.2±61.8

Values are mean ± S.D (n = 6 per group of which 3 males and 3 females). Control and treatment groups were compared statistically using one

way ANOVA followed by Dunnett’s test.

Table 10: Quantitative data on absolute organ weight of rats treated with Akil Kattai Chooranam for 28 days in Sub-acute toxicity study.

Group HEART

(gms)

LIVER

(gms)

KIDNEYS

(gms)

SPLEEN

(gms)

BRAIN

(gms)

LUNG

(gms)

STOMACH

(gms)

TESTES

(gms)

UTERUS &

OVARY (gms)

Group I 0.54±0.15 4.67±0.70 1.28±0.12 0.55±0.25 1.38±0.22 1.31±0.19 1.23±0.22 2.7±0.60 1.33±0.15

Group II 0.75±0.17 5.02±0.58 1.32±0.19 0.61±0.18 1.68±0.21 1.51±0.1 1.18±0.37 2.8±0.78 1.33±0.25

Group III 0.74±0.20 6.42±1.43 1.49±0.23 0.48±0.21 1.61±0.19 1.7±0.21 1.43±0.39 3.2±1.1 1±0.17

Values are mean ± S.D (n = 6 per group of which 3 males and 3 females) for Heart, Liver, Kidney, Brain, Spleen, Lung, Stomach. Values are

mean ± S.D (n = 3 per group per sex ) for testes , ovary and uterus for Control and treatment groups were compared statistically using one way

ANOVA followed by Dunnett’s test.


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Low Power Magnification 10X

GROUP I GROUP II GROUP III

High Power Magnification 40X


Figure 01: Histopathology of Brain (Male Rat) in Sub-acute toxicity Study.





Figure 02: Histopathology of Brain (Female Rat) in Sub-acute toxicity Study.


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Low Power Magnification 10X.




Figure 03: Histopathology of Heart (Male Rat) in Sub-acute toxicity Study.





Figure 04: Histopathology of Heart (Female Rat) in Sub-acute toxicity Study.


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Figure 05: Histopathology of Kidney (Male Rat) in Sub-acute toxicity Study.





Figure 06: Histopathology of Kidney (Female Rat) in Sub-acute toxicity Study.


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Figure 07: Histopathology of Liver (Male Rat) in Sub-acute toxicity Study.





Figure 08: Histopathology of Liver (Female Rat) in Sub-acute toxicity Study.


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Figure 09: Histopathology of Lung (Male Rat) in Sub-acute toxicity Study.





Figure 10: Histopathology of Lung (Female Rat) in Sub-acute toxicity Study.


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Figure 11: Histopathology of Spleen (Male Rat) in Sub-acute toxicity Study.





Figure 12: Histopathology of Spleen (Female Rat) in Sub-acute toxicity Study.


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Figure 13: Histopathology of Stomach (Male Rat) in Sub-acute toxicity Study.





Figure 14: Histopathology of Stomach (Female Rat) in Sub-acute toxicity Study.


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Figure 15: Histopathology of Testes (Male Rat) in Sub-acute toxicity Study.





Figure 16: Histopathology of Uterus (Female Rat) in Sub-acute toxicity Study.


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Figure 17: Histopathology of Ovary (Female Rat) in Sub-acute toxicity Study.

4. DISCUSSION

In modern society, toxicology has become an important element in environmental and

occupational health. This is because many organizations, governmental and non-

governmental, utilize information from toxicology to evaluate and regulate hazards in the

workplace and nonoccupational environment. As part of prevention strategies, toxicology is

invaluable, since it is the source of information on potential hazards in the absence of

widespread human exposures. Toxicological methods are also widely used by industry in

product development, to provide information useful in the design of specific molecules or

product formulations.[12]

NOEL (NOAEL) means the no observed (adverse) effect level, or the highest dose that does

not cause a toxic effect. To establish a NOEL requires multiple doses, a large population and

additional information to make sure that absence of a response is not merely a statistical

phenomenon. LOEL is the lowest observed effective dose on a dose-response curve, or the

lowest dose that causes an effect.[13]


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Herbal medicines have been used for thousands of years, and herb preparations are

commercially available for the folk remedies or for the promotion of health. In contrast to

chemical drugs, herbs are regarded to be non-toxic and safe, because of their natural origin.

But accumulating clinical data claim the toxicity of herbal medicine. The results of the acute

toxicity profiling of the drug AKC reveals no significant change in body weight, behavioral

and mortality at the dose of 2000mg/kg.

The liver is considerably the most important target for toxicity caused by drugs. This

susceptibility is a consequence of the functional features of the liver and their role in the

metabolic elimination of most drugs. Therefore, evaluation of potential hepatotoxicity

represents a critical step in the development of new drugs. The liver is very active in

metabolising foreign compounds and, although biotransformation reactions generally parallel

detoxification processes, the formation of reactive metabolites is relatively frequent. Thus,

drug-induced hepatotoxicity can be due to the administered compound itself or to metabolites

formed by hepatic metabolism.[14]

In sub-acute toxicity study treatment with AKC at the dose level of 200 and 400mg/kg did

not reveal any significant change in body weight, food and water intake in both male and

female rats. Further there is no alteration in AST, ALT and ALP level of treated animals

which denotes the hepato protective nature of the drug.

Blood a fluid connective tissue involved in supply of nutrient’s and also drug to the target

organs often exposed to the ill effect of certain cytotoxic drugs. Most of the siddha

preparation seems to be safe with hematopoietic system but certain drugs and chemicals

which particularly have tendency to disturb the bone marrow and causes hemolysis disturbs

the cardiovascular physiology. The hemo toxic nature of the drug exerted by the fluctuation

in blood cell count in particular to RBC and WBC cells. Low hemoglobin content reflects the

low level of RBC which in turn affects the oxygen carrying capacity of the blood. At the end

of the most of the toxicity studies the blood collected from the animal before sacrifice will be

subjected to whole blood analysis and also to serological analysis.

Results of hematological and serological profiling of AKC at the dose of 200 and 400mg/kg

did not shown any signs of hematopoietic toxicity and there is no change in the blood and

serum parameters of the treated animals.


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5. CONCLUSION

From the results of the present toxicology study of the trial drug AKC there was a generation

of evidence based data that shows the drug is relatively non-toxic, causes no apparent organ

damage or mortality in both the short term and long term administration on study animals.

Hence from the results, it was concluded that the drug AKC was safe and reveals no signs of

significant toxicity for long term treatment for the chronic disease condition.

6. REFERENCES

1. OECD. 2003. Descriptions of Selected Key Generic Terms Used in Chemical Hazard

Risk Assessment. Joint Project with IPCS on the Harmonisation of Hazard Risk

Assessment Terminology. OECD Series on Testing and Assessment No. 44. Environment

Directorate, Joint Meeting of the Chemicals Committee and the Working Party on

Chemicals, Pesticides and Biotechnology. ENV/JM/MONO (2003)15. Paris: OECD.

2. Rozman K. Disposition of xenobiotics: Species differences. Toxicol. Pathol, 1988; 16:

123–129.

3. Pyszel A. Effect of metals, benzene, pesticides and ethylene oxide on the haematopoietic

system. Med Pr., 2005; 56: 249-55.

4. Ioannides C, Lewis DF. Cytochromes P450 in the bioactivation of chemicals. Curr Top

Med Chem., 2004; 4: 1767-1788.

5. Jha V, Chugh KS. Nephropathy associated with animal, plant, and chemical toxins in the

tropics. Sem Nephrol, 2003; 23: 49–65.

6. Mohan RC, Agasthiyar Paripooranam 400. Thamarai Noolagam, Chennai-26, Edition-

2012: 111.

7. OECD guideline for testing of chemicals. Guideline 423, 17th December 2001.

8. OECD Guide lines 407 for testing of chemicals. Repeated dose 28-Day Oral Toxicity

Study in Rodents, 2008; 2- 8.

9. Jain N, Sharma P, Sharma N, Joshi S C. Haemato-biochemical profile following sub

acute toxicity of malathion in male albino rats. Pharmacologyonline, 2009; 2: 500–506.

10. Suvarna SK, Layton C, Bancroft JD. Bancroft’s theory and practice of histological

techniques. 7th edn, Churchill Livingstone, London, 2013.

11. Visweswara Rao. Biostatistics, A manual of statistic methods for use in Health, Nutrition

and Anthropology, Rajkamal Electrical press, Delhi, 2007; 226-312.

12. US department of Health and human services. Asbestos Toxicity. Case Studies in

Environmental Medicine, 2005.


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13. Wolfgang Dekant. Evaluation of Toxic Effects. Ullmanns Encyclopedia of Industrial

Chemistry, 2011.

14. Gomez-Lechon MJ. In Vitro Evaluation of Potential Hepatotoxicity Induced by Drugs.

Current Pharmaceutical Design, 2010; 16: 01-15.

Documents

SAFETY EVALUATION OF SIDDHA FORMULATION AKIL KATTAI