A novel microplate-based assay for screening radioprotectors and its validation based on DNA and membrane system

Ionizing radiation leads to damage at various cellular and sub-cellular levels and can be prevented by radioprotectors. There are many in vitro and in vivo but rather expensive assays for screening of radioprotectors from natural and synthetic sources. We have developed a cell free radioprotector screening assay which involves bleaching of crocin pigment, isolated from saffron by radiolytic products of water. Any molecules/compounds which can inhibit the bleaching of the crocin will act as a radioprotector. The developed assay was further validated by the existing in vitro assays. Different radioprotectors have different level for inhibition of bleaching of crocin. The trends of radioprotection offered by crocin bleaching assay, plasmid relaxation and lipid peroxidation are TMG > FA > VA > Amifos > Trox, TMG > VA > FA > Amifos > Trox, and TMG > FA > Trox > VA > Amifos, respectively. We are getting different trends for different assays. This is because different drugs have different mechanisms of radioprotection in different assay systems. In conclusion, the crocin bleaching assay developed here is a simple, fast and economical screening assay and it will have great value in radioprotection programme for screening many potential compounds for radioprotection.