Explaining Ayurvedic preparation of Rasasindura, its toxicological effects on NIH3T3 cell line and zebrafish larvae

Rasasindura is a mercury-based medicinal formulation that contains HgS (>99%). Although cinnabar ore was a well-known mineral in the past, the Ayurvedic practitioner adopted a critical and tedious procedure for the preparation of Rasasindura. Therefore, it is essential to understand the Ayurvedic process in the perspective of material science. Further, a toxicity study is also required as mercury is the main component in Rasasindura. Here, in the present study, we characterized Rasasindura and one of its intermediates (Kajjali) to understand the physicochemical changes that occur in the Ayurvedic process. Furthermore, we have assessed the toxicity of Kajjali and Rasasindura in NIH3T3 cell lines and zebrafish larvae. XRD analysis of Rasasindura confirms it as a highly pure α-HgS with size ranges from nano to micron sizes (starting from ∼80 nm). Whereas, Kajjali is a β-HgS having lower size ranges (starting from ∼30 nm). Rasasindura did not show significant cytotoxicity on NIH3T3 cell line up to 75 ppm, whereas for Kajjali, cytotoxicity was observed above 20 ppm. The higher toxicity of Kajjali is due to higher penetration of particles into the cells. However, in zebrafish larvae, even at too high concentrations (1000 ppm), both Rasasindura and Kajjali did not show any toxicity or morphological changes. This study concludes that Rasasindura is not toxic up to a reasonable concentration. Further, these two drugs did not contain toxic organic mercuric compound; otherwise, it could have been lethal to the zebrafish larvae.