Acute toxicity and sub-acute toxicity study of Siddha herbal formulation “Jaathipalathichooranam”

R.Subashini¹, A. Manoharan², G.Subash Chandran ^{3 1}

PG Scholar, Department ofPothuMaruthuvam, GSMC,Palayamkottai, Tamilnadu, India ²Professor and Head of the Department,Department of PothuMaruthuvam, GSMC, Palayamkottai, Tamilnadu, India ³Lecturer, Department ofPothuMaruthuvam, GSMC,Palayamkottai, Tamilnadu, India

Abstract

Background

Jaathipalathichooranam (JPC) is Siddha poly herbal preparation, primarily composed of Myristicafragrans(Linn). In terms of Pharmacological action JPC ishaving significant bronchodilator (Nisha et.al,2017)antispasmodic, antidiabetic and hypolipidemic actions.(Arulmozhi et.al,2007 ). As per the reference text Gunapadamporutpanpunool, the indication of JPC is prescribed primarily for the management of Respiratory disease , DUB and Vatha diseases (Murugesa Mudhaliyar,2016).

Objective

To determine the acute and sub-acute toxicity effect of the siddha poly herbal formulationJaathipalathiChooranam (JPC).

Methods

Acute &sub-acute toxicity of JaathipalathiChooranam wasevaluated in wistarrat models with oral administration of JPC 50mg/kgbw for seven days in acute and 20 days in subacute toxicity.All the studies were carried under OECD Guidelines.

Results

TheJaathipalathiChooranam has not produced any acute and sub-acute toxicity symptoms, no changes in corporal weight and haematological parameters and hepatic enzymes like SGOT & SGPT . The result of the study suggests that JPC is safe to use for long term prescription.

Conclusion

The results suggested that Jaathipalathichooranamis found to benon-toxic, when action of JPC was analyzed onhematopoietic and leucopoietic systems.Keywords

Jaathipalathichooranam , Siddha drug, Siddha medicine, Toxicity studies

Acute toxicity and sub-acute toxicity study of Siddha herbal formulation “Jaathipalathichooranam”

Introduction

The drug Jaathipalathichooranam is a poly herbal formulation, which has indicated for the treatment of Swasakasam (Bronchial asthma). It is a common and inflammatory disease, irreversible damages in bronchial tree. It is characterized by recurrent episode of wheezing, dry cough, chest tightness, and shortness of breath. These episodes may occur a few times a day or a few times per week.

Depending on the person they may become worse at night or with exercise.Bronchial asthma is a very common disease in the society due to increasing exposure to air pollution and western life style. It is common in both sex but more prevalent among boys, while during adolescence and adulthood, it affects girls and women more. In 2025, especting more than 300 million people were affected in worldwide. In future in India wills affected more than 100 million peoples have affected in this disease.It is one of the leading causes of morbidity and mortality in rural India (Smith 2000).

Materials and Methods

TheJaathipalathiChooranam (JPC) is a good expectorant, antispasmodic action, Immuno-modulator, Bronchodilator (Nisha et.al,2017), Anti-oxidant, Anti- hyperlipedemic, Anti-diabetes actions(Arulmozhiet.al,2007). It was mentioned in SarabendhraVaithiyaMuraikal- KasaSwasaRogaSikicha Siddha text book .(pg.no:131,132) Mr.K.Vasudevasasthri, Dr.S.Venkadarajan, L.I.M.(Retd.),year of edition-2006.

The study was carried out in the Department of Pharmacology with the approval of the Institutional Animal Ethics Committee, IAEC/R.SUBASHINI/TNMGRMU/MD(S)/321611009/KMCP/28/

2018 KM College, Tamilnadu

Collection of raw drug

The raw drug of JaathipalathiChooranam was purchased from Nagercoil based medical shop and authenticated by Medicinal Botanist &Gunapadam experts at Govt. Siddha Medical College, Palayamkottai-627002.

The combinations of JPC and Quanity is given below,

Methods of study

Animal studies

Healthy adult male and female Wistar albino rat weighing between 170-200 g were used in this study. The animals werefeed normal water and ad libitum. All the animals were acclimatized to the laboratory for 7 days prior to the start of the study. Acute oral toxicity of Jaathipalathichooranamis carried out as per OECD -423 guidelines. After the ethical clearance from Institutional Animal EthicsCommittee (KMCP/28/1.5.18).Determination of acute oral toxicity is usually the initial screening step in the assessment and evaluation of the toxic characteristics of all compounds. Acute toxicity is involved in estimation of LD₅₀ (ShettyAkhila, et al.2007).

The wistar rat were put on fastover night and provided only water, after which the Jaathipalathichooranam is administered orally50 mg.kg ^-1 body weight in Tween-80. The animals are then observed for 14 days and maintained with normal food. A mortality rate of 2 or 3 animals in 14 days is recorded and the dose is said to be toxic dose. But when mortality of one animal is observed, then the same dose is repeated again for confirmation. However, if mortality is not observed, the procedure is repeated for further higher doses such as 300 and 2,000 mg.kg^-1 body weight. Any occurrences of toxic symptoms were observed for 72 hrs(Shah Ayub, 1997, Bürger, 2005). All test animals were subjected to gross necropsy.

Sub-acute test for Jaathipalathichooranam(JPC)

This experiment evaluates thesub-acute toxicity potential of Jaathipalathichooranam. Male and female Wistar rats weighing 180 ± 10 g were used in this study. The animals were divided into five groups of six animals each. The GroupI were administered with a single daily dose of 0.5 ml of Tween 80 orally for 20 days. The animals in Group II wereadministered with 50 mg.kg^-1b.w. of the JPCorally once daily for 20 days. The animals in Group III were administered with 100 mg.kg^-1b.w. of the JPC orally once daily for 20 days. The animals in Group IV and V were administered once daily with 200 and 400 mg.kg^-1b.w. for 20 days (Pieme,et al 2006, Joshi, et al 2007, Mythilypriya, et al., 2007). The animals were then weighed every five days, from initial day of the treatment, to record the weight variation. The collected blood samples wasanalyzed in lipids,renal profiles and hepatic enzymes.

Results

Acute toxicity study of Jaathipalathichooranamhas showed no mortality or morbidity inover 15-days of period following single oral administration (Table-1). The animals did not showed any changes in the general appearance, physiological and pathological changes (Table no .1) when the trial drug was administered at the dose of 2000 mg/kg/bw.so the LD50 of Jaathipalathichooranamwas taken as2000 mg/kg/bw.

TABLE NO.1. ACUTE TOXICITY STUDY OF JAATHIPALATHICHOORANAM

ST- Sign of toxicity; NB- Normal Behaviour; D- Died; S- Survive. Values are expressed as number of animals (n=3).

SUB-ACUTE TOXICITY OF JAATHIPALATHICHOORANAM

The effects of Jaathipalathichooranamon body weight changes in rats

The effect of Jaathipalathichooranam(JPC) on the body weight changes of rat models were noted .from the study it was observed that, there was significant increase (p<0.05) in body weight in all the animals observed. The results are shown in Table.2. A study on the effects of Jaathipalathichooranamon body weight changes in rats was carried out. The values are expressed as mean ± S.E.M. n=6. The results of group I were compared with other groups such as II, III, IV, and V. The statistical analysis was carried out using one way ANOVA method, where **P<0.01 *P<0.05.

Table.2.The effects of Jaathipalathichooranamon body weight changes in rats

The effects of Jaathipalathichooranam kidney, heart, liver and brain of the rats

The effects of Jaathipalathichooranamon kidney, heart, liver and brain of the rats were observed. From the study it was clear that, significant (p<0.01) changes in the weights of various organs of the animals occurred with higher doses of the extract (400 mg.kg^-1 bwt), Macroscopic examinations did not show any changes in colour of the organs of the treated animals compared with the control. The results are shown in Table.3. The study on the effects of Jaathipalathichooranamon kidney, heart, liver and brain of the rats was tested. The values are expressed as mean ± S.E.M. n=6. The results of group I were compared with other groups such as II, III, IV, and V. The statistical analysis was carried out using one way ANOVA method, where **P<0.01.

TABLE.3.THE EFFECTS OF JPC KIDNEY, HEART, LIVER AND BRAIN OF THE RATS.

Effect of Jaathipalathichooranam on biochemical profiles

The effect of Jaathipalathichooranamon various biochemical value showed a significant decrease (p<0.05) in the plasma glucose level at higher dose (400 mg.kg^-1) compared with control rats. Significant decrease (p<0.05) in the plasma total cholesterol (TC), triglyceride (TG) and LDL-cholesterol levels. But a significant increase (p<0.05) in HDL-cholesterol levels were observed in treated animals, it was compared with control groups. AST, ALT and ALP levels were also normal in the JPC treated animals. such as glucose, cholesterol, triglyceride, HDL and LDL in rats was tested where, group I animals (GPI) The values are expressed as mean ± S.E.M. n=6. The results of group I were compared with other groups such as II, III, IV, and V. The statistical analysis was carried out using one way ANOVA method, where **P<0.01 *P<0.05 (Table no4.).

TABLE.4.THE EFFECT OF JAATHIPALATHI CHOORANAMON BIOCHEMICAL PARAMETERS SUCH AS GLUCOSE, CHOLESTEROL, TRIGLYCERIDE, HDL AND LDL.

Changes in hepatic enzymes .

According to Table no 5 showed no elevation in hepatic enzymes level. The group I animals (GPI) were treated with normal saline (5ml.kg^-1 ), group II animals (GPII) with 50 mg.kg^-1 of HAEBD group III animals (GPIII) with 100 mg.kg^-1 ofJaathipalathichooranam, group IV animals (GPIV) with 200 mg.kg ^-1 of Jaathipalathichooranam, and group V animals (GPV) with 400 mg.kg^-Jaathipalathichooranam. The values are expressed as mean ± S.E.M. n=6. The results of group I were compared with other groups such as II, III, IV, and V. The statistical analysis was carried out using one way ANOVA method, where **P<0.01 *P<0.05.

Table.5.The effects of Jaathipalathichooranamon biochemical parameters such as AST, ALT, ALP, TP and Albumin in rats

Effect of Jaathipalathichooranam on haematological parameters in rats

The effects of JPC observed for its effect on hematological parameterson the experimental rats. From the study it was evident that, a significant increase (p<0.01) were observed in the hemoglobin contents and RBC count in the group treated with 200 mg.kg^-1 body weight of JPC and a significant decrease of the parameters occurred in the group treated with 400 mg.kg^-1b.w.t compared with the control. There was no significant change in the calcium level in all the treated animals compared to the control (Table no.6). The statistical analysis was carried out using one way ANOVA method, where *P<0.05.

TABLE.6. THE EFFECT OF JAATHIPALATHI CHOORANAM ON HAEMATOLOGICAL PARAMETERS

Discussion

The evaluation of acute and sub-acute toxicity in experimental animals is relevant in determining the overall toxicity of the herbal preparation. The acute toxicity study of JPC was not produced at the dose of 2000 mg/kg/bw.Hence, 1/10^th of 2000 mg.kg^-1 i.e. 200 mg.kg ^-1 of dose was selected as a minimum dose for sub-acute toxicity study (Abu TahaNael, et al., 2008).(8). The result of sub-acute toxicity study shows that there was no significant change in animal behaviour due to the absence of toxicity. The animals treated with JPC had normal growth pattern and body weight when compared with control rats treated with normal saline. The JPC at all concentrations do not produce liver damage. There were no significant changes in the hematological parameters between control and treated groups even whenJPC was administered with higher dose of 400 mg.kg ^-1.

Conculsion

According to the present study report JPCis non-toxic &safer to usein long duration. While concluding the biochemical and Liver enzymes studyreports, no evidence of severe toxicity was associated with the administration of higher concentration of Jaathipalathichooranam.

References

1. Abu TahaNael, A., Alkhawajah, M., Aziz Raveesha, K.K., 2008. Acute and subacute toxicity studies of PerseaamericanaMill (Avocado) seed in rats. International Journal of Medical Toxicology and Legal Medicine 11 (2), 10-16.

2. Adeneye, A.A., Ajagbonna, O.P., Adeleke, T.I., Bello, S.O., 2006. Preliminary toxicity and phytochemical studies of the stem bark aqueous extract of Musangacecropioidesin rats. Journal of Ethnopharmacology 105, 374-379.

3. D.k.Arulmozhi,R.Kurian,A.Veeranjaneyulu&S.L. Bodhankar. Antidiabetic and antihyperlipidemiceffects of Myristicafragrans.in animal models,pharmacologicalbiology,7oct2008.

4. Bürger, C., Fischer, D.R., Cordenunzzi, D.A., Batschauer de Borba, A.P., Filho, V.C., Soaresdos Santos, A.R., 2005. Acute and subacute toxicity of the hydroalcoholic extract from Wedeliapaludosa (Acmelabrasilinsis) (Asteraceae) in mice. J. Pharm. Sci. (www.cspsCanada.org) 8(2):370-373

5. Hayes, A.W., 1989. Guidelines for acute oral toxicity testing. In: Principles aand Methods of Toxicity. New York: Raven Press Ltd, 184

6. Murugesamudaliar, GunapadamMooligaivaguppu, 2^st Edition,

Department of Indian Medicine and Homeopathy, 2016

7. Mythilypriya, R., Shanthi, P., Sachdanandam, P.,2007. Oral acute and subacute toxicity studies with Kalpaamruthaa, a modified indigenous preparation on rats. J. Health Sci. 53(4): 351-358

8. Nadkarni, K.M., 1976. Indian Material Medica. Popular Prakashan Pvt. Ltd.,Bombay,pp:1202-1211

9. R.Nishayadav ,A.Kingsly, G.Esakipandian,R.AntonyDurachi, Bronchodilator Activity Of SwasakudoriChooranam.International Journal Of Current Research In ChemistryAndPharmaceuticlSciences,

10. Olson, H., Betton, G., Robinson, D., Thomas, K., Monro, A., Kolaja, G., Lilly, P., Sanders, J., Sipes, G., Bracken, W., Dorato, M., Deun, K.V., Smith, P., Berger, B., Heller, A., 2000. Concordance of toxicity of pharmaceuticals in humans and in animals. Regulatory Toxicology and Pharmacology 32, 56–67

11. Pieme CA, Penlap VN, Nkegoum B, Taziebou CL, Tekwu EM, Etoa FX, Ngongang J (2006). Evaluation of acute and subacute toxicities of aqueous ethanolicextractof leaves of (L) Roxb(Ceasalpiniaceae). Afr. J. Biotechnol. 5(3): 283-

12. Raza, M., Al-Shabanah, O.A., El-Hadiyah, T.M., Al-Majed, A.A., 2002. Effect of prolonged vigabatrin treatment on haematological and biochemical parameters in plasma, liver and kidney of Swiss albino mice. ScientiaPharmaceutica, 70, 135-145.

13. ShettyAkhila. J., Shyamjith, Deepa , Alwar, M.C., 2007.Acute toxicity studies and determination of median lethal dose Current science 93,7, 917.

14. Shah Ayub, M.A., Garg, S.K., Garg, K.M.,1997. Subacute toxicity studies on Pendimethalin in rats. Indian J. Pharm. 29: 322-324.

15. Tofovic, S.P., Jackson, E.K., 1999. Effect of long-term caffeine consumption on renal function in spontaneously hypertensive heart failure prone rats. Journal of Cardioavascular Pharmacology, 33, 360-366.

16. Teo, S., Stirling, D., Thomas, S., Hobermann, A., Kiorpes, A., Khetani, V., 2002. A 90- days oral gavage toxicity study of D-methyl penidate and DL methyl penidate in Sprague-Dawley rats. Toxicology, 179, 183.