MICROWAVE ASSISTED EXTRACTION OF BIOLOGICAL COMPONENTS OF CROCUS SATIVUS WITH DNA BARCODING TECHNOLOGYAbstract
Saffron is derived from the flower of Crocus sativus as a dried stigma. Kashmiri saffron is the most expensive herb and in sky-scraping demand by herbal and pharmaceutical industries due to which the illegal trade is prevailing in the industry to earn instant Profit. Detection of adulteration and comparison of genuine saffron vs. fake saffron is the crucial aspect which needs a scientific solution. Samples were taken Mogra (A), Lacha (B), Iranian (C) and unknown grade of saffron (D) from Pampore and Anantnag region of Kashmir. The DNA from the samples was extracted with the help NucleoSpin® Plant II Kit and were amplified with the help of reported universal primers of matk, rbcL and MatK, and bidirectional Sanger sequencing was performed. Only ITS gene showed positive extraction and amplification results. ITS gene was capable of distinguishing the interspecific as well as intraspecific variations. The blind trial conducted on unknown grade sample revealed the presence of substituent, which turned out to be Safflower, i.e. Carthamus tinctorius as observed through nucleotide blast search. The site-specific variations were observed, and the nucleotide sequences of ITS obtained during the study were submitted to the NCBI gene bank with accession number MH984846, MH984847, and MH984848. The examination of saffron with DNA Barcoding methodologies may contribute to health care communities a new validated tool to authenticate the samples and improve the efficacy of herbal medicine used to treat various ailments.
T. Sharma *and M. K. Thakar
Department of Forensic Science and Toxicology, Department of Forensic Science, Chandigarh University, Gharuan, Punjab, India.
12 October 2018
21 June 2019
26 June 2019
01 July 2019