Extraction and Purification of Coumarins in <i>Ipomoea cairica</i> by TLC and HPLC: An Integrative Experiment for Student Learning Chromatography

Chromatographic techniques played a major role in several separation processes. In this work, an integrative laboratory practice is proposed in which students are expected to isolate scopoletin and umbelliferone from raw Ipomoea cairica. Thin layer chromatography (TLC), preparative chromatography (PC), and high-performance liquid chromatography (HPLC) techniques are applied to isolate phytochemical bioactive compounds. Scopoletin and umbelliferone are coumarins that have been reported to possess anti-inflammatory and antioxidant properties. These metabolites are found in high concentrations in Ipomoea cairica stems, and their methanolic extracts have shown larvicidal properties against Aedes aegypti. The experimental procedure includes fundamental extraction techniques such as liquid extraction and ultrasonic extraction. Additionally, it incorporates more advanced methods, such as chromatographic separation on a thin-layer plate, analytical detection of coumarins using HPLC with a diode array detector, and the isolation of coumarins through preparative plate chromatography. This laboratory experiment allows students to integrate and compare, in two laboratory sessions, the basic principles of TLC and preparative chromatography with more advanced techniques such as HPLC.