Toxicological study of Siddha Drug "Kalludaikkudoori Mathirai”

The ingredients of KALLUDAIKKUDOORI MATHIRAI were purified and the drug was prepared according to the process mentioned in SIDDHA VAIDHIYA THIRATTU (Pg. no: 47-48, Edition-2014, K.n. Kuppusamy mudhaliar and K. S. Utthamarayan). The drug was selected for evaluating the toxic effect and mortality when given in short and long duration. The main aim of this study is to access the safety of the siddha drug “KALLUDAIKUDORI MATHIRAI” on wistar albino rats under various dose levels of drug administration especially in acute and subacute toxicity studies. In review of literature, the ingredients of KALLUDAIKKUDOORI MATHIRAI were discussed in depth with a special attention paid to their medicinal uses and toxicological aspects. The ingredients of KALLUDAIKKUDOORI MATHIRAI are rasam, gandhagam and siruganpeelai samoolam. The rasam and gandhagam purchased from registered raw material supply shop – Rajendra herbal store, Thuckalay, Kanyakumari District and Siruganpeelai collocted from Ammapet. The raw samples were taken for purification and test medicine was prepared, as per the method narrated in the literature. Phytochemical study of KALLUDAIKKUDOORI MATHIRAI indicates the presence of Alkaloids, steroids, Triterpenoids, Phenol, Tannin, Saponins, Sugar, Betacyanin. FTIR study of KALLUDAIKKUDOORI MATHIRAI shows the presence of functional groups such as Primary, secondary, amines, amides, Alkenes, Alkynes, Nitro compounds, Aromatics, Alkanes, Alcohols, carboxylic acids, esters, ethers, Aliphatic amines, Alkyl halides. ICP OES results shows the formulation of KALLUDAIKKUDOORI MATHIRAI is safe as it contains heavy metals within specified limits. It also has physiologically important minerals like calcium, iron, mercury, Pottassium, Sodium, Phosphorus, Sulphur. In KALLUDAIKKUDOORI MATHIRAI the heavy metals like Arsenic, Cadmium, Nikkal and Lead were below detectable level. This reveals the safety of the drug and it has free from toxic substances and has no side effects. GCMS Study of Kalludaikkudoori mathirai shows the compounds presence of Antimicrobial, antifouling, antibacterial, cancer preventive, hyper-cholesterolemic, 5-alpha reductase inhibitor, anti-androgenic, perfumery insectifuge, anti-inflammatory, anemiagenic, dermatigenic, choleretic, anti-phlogistic, anti-melanoma activity, antioxidant, diuretic activity and antifungal activities XRD is used for analysis of the amount of the major classes of active constituents present in drug of KALLUDAIKKUDOORI MATHIRAI. X-ray powder diffractometry (XRD) is used to analyse different minerals, crystalline materials and metallic based herbal formulations. The herbo mineral drug KalludaikkudooriMathirai was estimated by XRD and the intense sharp diffraction peaks (21, 28, 33, 44, 46, 53, 58, 63, 68, 76.) clearly confirmed the presence of crystallinity. X-ray powder Diffraction data confirmed the formation of herbo organic complex molecules. HPTLC finger printing analysis of the KALLUDAIKKUDOORI MATHIRAIreveals the presence of one prominent peaks corresponds to presence of one versatile phytocomponents present with in it with the Rf value of the 0.86. Further the peak occupies the major percentage of area of 100 % which denotes the abundant existence of such compound. The Biochemical analysis Siddha trial drug KALLUDAIKKUDOORI MATHIRAI which shows the presence of Sulphate, chloride, starch, Ferros iron, unsaturated compound, and Amino acid in it. The Biological screening (Aflatoxins& Pesticide residue) reveals results of the KALLUDAIKKUDOORI MATHIRAI shows the Aflatoxins B1, B2, G1, G2 are not detected. And then Organo chlorine pesticide Alpha BHC, Beta BHC, gamma BHC, Delta BHC, DDT and endosulphan are not detected. The Organo Phosphorus Malathion, Dichlorovus and chlorpyriphos are not detected. The PyrethroidesCypermethrin are not detected. In acute toxicity study all the animals were active and did not showed any signs of toxicity. The motor activities were normal in all the 5 groups of animals. This acute toxicity study results reveals that KALLUDAIKKUDOORI MATHIRAI was nontoxic upto a dose level of 2000 mg/kg body weight of animal. The acute toxicity study was conducted to know single dose toxicity of KALLUDAIKKUDOORI MATHIRAI on female Wistar albino rats. The female animals were selected for study of 6 weeks old with weight range of within ±20% of mean body weight at the time of randomization. The groups were numbered as group I,II,III,IV and V and dose with control, 5 mg/kg, 50 mg/kg, 300 mg/kg and 2000 mg/kg of KALLUDAIKKUDOORI MATHIRAI. The drug was administrated by oral route as single dose and observed for 14 days. Daily the animals were observed for clinical signs and mortality. Body weight of animals were recorded once in a week. There were no physical and general behavioural changes observed in wistar albino rats of 5 mg/kg, 50 mg/kg, 300 mg/kg and 2000 mg/kg to rats during 14 days. Body weight of all animals did not reveal any significant change as compared to vehicle control group. Food consumption of all group animals were normal. Mortality was not observed in all treated groups. In sub – acute toxicity animals were selected randomly grouped into four different groups containing minimum (5 male + 5 female) per groups. The groups were numbered as group I, II, III and IV dose with control, 13 mg/kg (low dose), 65 mg/kg (mid dose) and 135 mg/kg (high dose) of KALLUDAIKKUDOORI MATHIRAI. The KALLUDAIKKUDOORI MATHIRAI was administered doses once daily for 28 days and all animals were observed daily once. These observations were also performed on weekends. The observations included clinical signs of toxicity, food intake, water intake, body weight. No signs of toxicity were observed. There was no significant changes in food intake, water intake and body weight. No mortality occurred till the last day of the study. The blood samples are used to evaluate haematological parameters (like SGOT, SGPT and ALP). No changes in haematological parameters and biochemical parameters in low, middle and high dose (13 mg/kg, 65 mg/kg, 135 mg/kg). On completion of the 28 days of drug administration, wistar albino rats were sacrificed. In macroscopic examination the heart, kidneys, liver, brain and spleen were weighted. The organs were normal when compared with control group. Histopathological examination revealed normal architexture in comparison with control and treated animal in low, middle and high doses (13 mg/kg, 65 mg/kg, 135 mg/kg). CONCLUSION: KALLUDAIKKUDOORI MATHIRAI was studied for its acute and sub - acute toxicity effect by using laboratory animals. In acute toxicity study KALLUDAIKKUDOORI MATHIRAI did not produce any specific toxicity or mortality even upto the dose of 2000 mg/kg in rat. In sub – acute toxicity study, 13 mg/kg, 65mg/kg and 135 mg/kg of kalludaikkudoori Mathirai was used and it was administered once daily for 28 days through oral route. Kalludaikkudoori Mathirai did not alter the body weight and food intake. Water intake is higher in low dose, mid dose and high dose compared to control group. After 28 days the blood was collected and subjected to haematological parameters. Kalludaikkudoori Mathirai at 13 mg/kg, 65 mg/kg and 135mg/kg was found to be safe and did not alter any of the biochemical parameters, Haematology and Histopathology (Liver, Kidney, spleen and brain) during my study period. From the study it was concluded that, low, middle and high doses (13 mg/kg, 65 mg/kg and 135 mg/kg) of Kalludaikkudoori Mathirai was found to be safe when administered orally in human for long time. Finally the dose mentioned in literature is safe for clinical use