|Year : 2016 | Volume
| Issue : 1 | Page : 39-43
Toxicity studies of iron-containing ayurvedic drug Kasisa Bhasma
Satadru Palbag1, Dhiman Saha2, Dev Nath Singh Gautam3
1 Department of Rasa Shastra, BIPS, SPLPIM Campus, Kalyani, Nadia, West Bengal, India
2 Department of Ayurvedic Pharmacy, RGSC, Banaras Hindu University, Barkachha, India
3 Department of Rasa Shastra, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India
|Date of Submission||28-Mar-2016|
|Date of Acceptance||17-May-2016|
|Date of Web Publication||2-Jun-2016|
Dev Nath Singh Gautam
Department of Rasa Shastra, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh
Source of Support: None, Conflict of Interest: None
Background: Kasisa is an important iron-containing mineral drug of ayurvedic Rasa Shastra and employs for various therapeutic uses in anemia, hair growth modulator, eye disorders, skin disorders, etc.
Objective: In this study, shodhana was done by triturating in lemon juice and calcination was done by traditional heating arrangement with Kanji (sour gruel) method. Both the purified and calcined product of the Kasisa was subjected to histopathological and toxicological study to evaluate the safety of this iron-based ayurvedic medicine.
Materials and Methods: Thirty-two adult Charles Foster albino rats of either sex, 16 for each drug (Shodhita Kasisa, Kasisa Bhasma), were taken for the whole study. Toxicological study of the brain, liver, kidney, and spleen was performed.
Results: Kasisa Bhasma at higher dose of 100 mg/kg showed some adverse effects in isolated organs of experimental animals, but the extent of damage is minimal compared to Shodhita Kasisa which showed adverse effects at 25-50 mg/kg.
Conclusions: Kasisa Bhasma is nontoxic and safer as compared to Shodhita Kasisa. It can be administered at a controlled dose to affect therapeutic efficacy.
Keywords: Bhasma, Bhavana, Kasisa Shodhana
|How to cite this article:|
Palbag S, Saha D, Gautam DS. Toxicity studies of iron-containing ayurvedic drug Kasisa Bhasma. BLDE Univ J Health Sci 2016;1:39-43
Rasa Shastra is a well-established branch of Ayurveda serving humanity with its unique heritage of drugs derived from metal, mineral, and animal origin combined with certain herbs. Different alternative systems of medicines, including Ayurveda, make use of herbal preparations for their curative effects. Shodhana (~purification) is an ayurvedic technique necessary for almost all kinds of drugs to remove their impurities or toxic contents and to improve its efficacy. Marana (~calcination) is another important ayurvedic process used to convert the ayurvedic minerals and metals in their calcined form by special heating arrangement known as puta. The "bhasma0" based on incineration of metals by traditional puta (quantum of heat) system which reduces the metals and ultimately nullifies its toxicity, increases bioavailability by reducing the particle size converting it into bhasma form. Thus, minerals or metals in the form of bhasma form become easier to get absorbed into the systemic circulation. According to Ayurveda, Kasisa is Kesya (~hair growth modulator), raktavardhak (hematinic), carminative, digestive, and good for eyes.  However, for proper use, it is essential that raw Kasisa is subjected to shodhana process and marana process. Ayurvedic processes such as shodhana and marana are necessary to remove their impurities or toxic contents and to improve its efficacy. To confirm this belief, the toxicological study and histopathological study of some organs of adult Charles Foster (C.F.) albino rats of either sex (150-210 g) were used before and after administration of Shodhita Kasisa and Kasisa bhasma.
In this study, Kasisa is purified with lemon juice by bhavana (drying through trituration) process and marana was done by traditional puta.
| Materials and Methods|| |
Shodhana of Kasisa
Raw Kasisa (250 g) was procured from Ayurvedic Pharmacy, Institute of Medical Sciences, Banaras Hindu University, Varanasi. Shodhana of Kasisa was done by giving bhavana with Nimbu Swarasa taking the sample in a mortar and then triturating it until the sample gets completely dried.  Since Kasisa is hygroscopic in nature, it takes almost 3 days to dry completely. It was pale greenish, very fine powder, soft, smooth to touch, and a little sandy in appearance. Two hundred and fifty grams of raw Kasisa was taken for study, and after Shodhana process, 180 g of Shodhita Kasisa was obtained yielding a loss of 28%. Percentage of moisture content in raw Kasisa was 8.8% and Shodhita Kasisa was 9.4%. Higher percentage of moisture content is due to the fact that the research work was done on August during rainy season.
Marana of Kasisa
Kasisa was taken in mortar and pestle, and Kanji (sour gruel) was added slowly until the powder gets completely immersed in the liquid. It was then subjected to the process of bhavana (levigation) and the process continued till the liquid got completely dried. The process was repeated seven times until the entire mixture became semisolid then round-shaped pellets were formed, arranged in an earthen crucible. After this, the crucible in which the pellets were kept was closed by another crucible, and the joint was sealed with cloth and clay and then dried. Sealed crucible was subjected to traditional puta system of heating. The puta was adopted by conventional method using Laghu puta to produce a temperature of around 650°C. A pyrometer is arranged to keep a note on the rise and fall of temperature. After self-cooling, the crucible was taken out, cleaned, and opened carefully. Pellets were collected and powdered to give brownish-red Kasisa bhasma initially. The powdered bhasma was again taken in mortar and pestle and levigated with lemon juice until the entire mixture got semisolid. Pellets were prepared, sun dried and again bhasmikaran was done using Laghu puta. Usually, the process was repeated until the sour taste of Kasisa got nullified by repeated action of heat. In this particular study, the Kasisa bhasma became tasteless after the above process was repeated for three times. After this light, bright red-colored bhasma free from all blemishes was obtained. Moisture content of Kasisa bhasma was 0.1%.
C.F. rats of either sex (150-210 g) aged between 4 and 6 months were used for the study. Animals were procured from the Central Animal House, Institute of Medical Sciences, Banaras Hindu University. Ethical clearance was obtained from the Central Animal Ethical Committee (No. Dean/13-14/CAEC/209). Totally, 32 (16 males and 16 females) rats, 16 for each drug (Shodhita Kasisa, Kasisa Bhasma), were taken for the whole study. They were divided into four groups containing four rats (2 males, 2 females) in one cage. The grouping was done based on 3 different concentrations of the therapeutic dose given to the rats for the entire study period. The rats were maintained the same concentration of the drug, i.e., control (3% gum acacia suspension), 25, 50, 100 mg of the drug/kg body wt. given twice daily from the start of the experiment. The dose was selected according to the U.S. Food and Drug Administration, Centre for Drug Evaluation and Research guidelines. Both the drugs (Shodhita Kasisa and Kasisa Bhasma) were prepared in 3% gum acacia solution and were administered through oral route. The subchronic toxicity study was continued for 28 days at par with pharmacological observations according to the OECD-407 guidelines.  On the 29 th day, the animals were sacrificed. Organs, namely, brain, liver, kidney, spleen, heart, lungs, and ovaries/testis were isolated and preserved in 10% formalin. For histopathological study, grossing of the organs was done by trimming and subjecting for dehydration in ascending grades of alcohol for 12-14 h. After dehydration, the tissues were embedded in melted paraffin, and blocks were prepared and fixed on small wooden cubes. The tissue sections were cut very fine at 3-5 microns. Tissue sections were stained with hemotoxylin and eosin. Finally, the sections were examined to observe histopathological changes on all collective tissues. The toxicity study was performed at par with the hematological study carried on the same experimental animals and the findings such as histopathological changes in the body and sub-chronic toxicity to specific organs were noted down. ,
| Results and Discussion|| |
The effect of Shodhita Kasisa and Kasisa bhasma on the weight of vital organs was measured following daily oral administration at different doses for 28 days [Table 1] and [Table 2]. No significant changes were observed in normal anatomy of the skin, hair, etc. Two mortalities were found in rats given Shodhita Kasisa at higher dose. Both were female rats. One of them died at 2 nd week of the study when given a dose of 5 mg/100 g body wt. and other died on the 3 rd week of the study when given a dose of 10 mg/100 g body wt. Histopathological changes were studied in the following organs, namely, brain, liver, kidney, and spleen of the rats after they were exposed to the test drug given in various concentrations for 28 days [Table 3] and [Table 4]. Kasisa Bhasma is nontoxic and safer compared to Shodhita Kasisa at higher dose and did not show any significant change in normal behavior of animals. There is no significant difference in weight of the vital organs. Shodhita Kasisa showed pathological changes starting from 25 mg/kg in the liver by projecting mild-minimal portal tract inflammation and mild sloughing of the tubular epithelial cells in the kidney. At 50 mg/kg dose of Shodhita Kasisa, pathological changes occurred on all the four organs, namely, brain, liver, kidney, and spleen. Whereas at a 25 mg/kg dose range of Kasisa bhasma, there is no significant changes observed. At 50 mg/kg of Kasisa bhasma, slight changes were observed in the liver and kidney. Toxicity study of Kasisa Bhasma at higher dose of 100 mg/kg showed some pathological changes such as vacuolation in hepatocytes and loss of hepatic architecture in the liver and slight decrease in cellularity of Purkinje cells was observed in the brain. Hence, although the toxicity study of Kasisa Bhasma at higher dose of 100 mg/kg showed some pathological changes in isolated organs of experimental animals, the extent of change was minimal compared to Shodhita Kasisa. As the main therapeutic role of Kasisa is hematinic and projects in the treatment of anemia, which is more common in female population, thus, it creates a doubt in our mind due to the mortality of female rats during the study that whether Shodhita Kasisa at higher dosage is safe for women. Shodhita Kasisa showed inflammation and necrosis to the liver and kidney at a dose of 25 mg/kg during histopathological studies. The toxicity data revealed the efficacy of bhasma and established the fact that metals and minerals are preferred in their bhasma form in Ayurveda to avoid toxicity and to have enhanced therapeutic potency [Figure 1],[Figure 2],[Figure 3],[Figure 4],[Figure 5],[Figure 6],[Figure 7] and [Figure 8].
|Table 1: Effect of Shodhita Kasisa on weight of vital organs (g) in rats measured weekly following daily oral administration at different doses for 28 days|
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|Table 2: Effect of Kasisa Bhasma on weight of vital organs (g) in rats measured weekly following daily oral administration at different doses for 28 days|
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| Conclusions|| |
Kasisa is an iron-containing mineral. Developing nations such as India where most of the rural female citizens suffer from mal-nutrition and anemia, indigenous cheap drugs such as Kasisa could be used to alleviate such diseases. Thus, ayurvedic drugs could play a major role in the National Rural Health Missions. Strategy should be developed to incorporate such drugs with proper validation. The above study is a preliminary approach to validate the Kasisa, both in its purified form and bhasmas form. The study proves the safety of Kasisa in its bhasma form at a regulated dose. We can conclude that Kasisa bhasma is safer in comparison to Shodhita Kasisa from the subchronic toxicological studies. Clinical studies and trial should be conducted over human beings to make more comprehensive investigation about the safety of Kasisa bhasmas. Further extensive research is welcomed in this arena.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Misra GS. Acharya Madhava′s Ayurveda Prakasa. Varanasi: Chaukhambha Bharti Academy; 2007. p. 325.
Jha CB. Ayurvediya Rasa Shastra. Varanasi: Chaukhabha Surbharati Prakasan; 2011. p. 252.
Sadananda S. Rasa Tarangini. 11 th
ed. Varanasi: Motilal Banarasidas Publication; 1979. p. 568.
OECD. Repeated dose oral toxicity test method. In: OECD Guidelines for Testing of Chemicals, Test No.: 407. Organization for Economic Cooperation and Development. Paris, France: OECD; 2008.
Ghosh MN. Toxicity studies. In: Fundamental of Experimental Pharmacology. 2 nd
ed., Ch. 26. Calcutta: Scientific Book Agency; 1984. p. 161.
Lewis SM, Bain BJ. Bates Dacie and Lewis. Practical Hematology. In: Bain JB, Bates I, editors. Basic Hematological techniques. 9 th
ed. London: Churchill Livingstone (Harcout Publishers LTD), NY; 2002. p. 19-46.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8]
[Table 1], [Table 2], [Table 3], [Table 4]