Microscopic, Macroscopic, and DNA Barcoding Techniques for Authentication of Crude Herbal Drugs

The rise of herbal medicine around the world has made it even more important to accurately identify crude herbal remedies to make sure they are safe, effective, and of good quality. This study used a combination of macroscopic, microscopic, and DNA barcoding techniques to verify twelve commonly used crude herbal medication samples from the Ayurvedic and Unani systems. The first visual and anatomical confirmation came from macroscopic and microscopic studies, respectively. DNA barcoding with rbcL and matK primers gave exact species-level identification by aligning the sequences with NCBI-BLAST. Five of the studied samples were consistently confirmed by all methods, but Bacopa monnieri was still doubtful because its shape was unclear and it only partially amplified. Using Cohen's Kappa (κ = 0.812), a statistical test, we found that the three methods agreed strongly with each other. These results show that a multi-tiered authentication strategy is better than using only one method. This supports the idea of a standardised protocol for validating herbal drugs in research and industry contexts.