ABSTRACT. DNA barcoding is a desirable tool for medicinal product authentication. DNA barcoding is a method for species identification using short DNA sequences that are conserved within species, but variable between species. Unlike animals, there is no single universal DNA barcode locus for plants. Coding markers, matK and rbcL, and noncoding markers, trnH-psbA (chloroplast) and ITS2 (nuclear), have been reported to be suitable for the DNA barcoding of plants with varying degree of success. Sixty-four accessions from 20 species of the medicinal plant Cassia were collected, and analyzed for these 4 DNA barcoding markers. PCR amplification was 100% successful for all 4 markers, while intra-species divergence was 0 for all 4 Cassia species in which multiple accessions were studied. Assuming 1.0% divergence as the minimum requirement for discriminating 2 species, the 4 markers could only differentiate 15 to 65% of the species studied when used separately. Adding indels to the divergence increased the percentage of species discrimination by trnH-psbA to 90%. In 2-locus barcoding, while matK+rbcL (which is recommended by Consortium for the Barcoding of Life) discriminated 90% of the species, the other combinations of matK+ITS and rbcL+trnH-psbA showed 100% species discrimination. However, matK is plagued with primer issues. The combination of rbcL+trnH-psbA provided the most accurate (100% species ID) and efficient tiered DNA barcoding tool for the authentication of Cassia medicinal products.

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The present investigation aims at the extraction, phytochemical constituent analysis and antioxidant activity analysis of Cassia angustifolia. The solvents such as water, Ethanol-water, Ethyl acetate and Methanol were utilized to optimize extraction process arrive at extract with higher yields and better antioxidant potency. It was observed that water provides highest yield among all the solvents (20.7%). Ethanol water also yielded almost same extraction yield (20.4%). Whereas, methanol provided 17% and ethyl acetate about 11% of the yield. It was observed that the methanol extract was found to have highest percent of polyphenols of 30% and Ethyl acetate extract was found to have about 23%. It was found that water-ethanol extract was proved to have highest contents of flavonoids of 15%. This is followed by water extract (13%). The results of DPPH free radical inhibitory activity studies indicate that water and ethyl acetate extracts were found to exhibit IC 50 value of 274 µg/ml and 288 µg/ml respectively. Both the extracts have proved to be moderately active at inhibiting the activity of DPPH. The other extract such as ethanolwater and methanol extracts were proved to be have IC 50 values of 369 µg/ml and 380 µg/ml respectively. Thus, the present investigation has provided crucial information on the phytochemical contents and antioxidant activities of extracts of Cassia angustifolia. The results warrant for further studies in this direction leading to isolation and characterization of individual molecules responsible for the antioxidant activities of the extracts from very common plants that are available in our neighborhood.