HPTLC Analysis for the Simultaneous Estimation of Caffeic Acid, Gallic Acid and Quercetin in Two Polar Fractions of S. cumini L. Leaves

The worldwide reliance on traditional medicinal plants has grown exponentially over recent decades. The leaves of S. cumini are rich in various bioactive components, such as flavonoids, tannins, alkaloids, glycosides, and anthocyanins, which are responsible for their therapeutic benefits. For the first time, the HPTLC method was established simultaneously to quantify phenolic compounds (gallic acid and caffeic acid) as well as the flavonoid quercetin in S. cumini L. leaves. Phenolic compounds and flavonoids are commonly utilized as antioxidants, antiinflammatory agents and natural food colorants. To achieve effective separation in HPTLC, a mobile phase of ETA: toluene: formic acid (2:1:7, v/v) was used, with flavonoids as well as phenolic compound analysis performed on F254 TLC plates. Ultraviolet absorbance was observed at 254nm, 300nm, and 325nm for quercetin, gallic acid, and caffeic acid, respectively. The calibration curve for all three compounds exhibited linearity within the the100-900ng per spot range, with r² values of 0.9481, 0.9811, and 0.9828 for caffeic acid, gallic acid, and quercetin, respectively. During analysis, in the ethanolic fraction, caffeic acid (2.776), gallic acid (0.208) and quercetin (0.129) were found, whereas in the water fraction, caffeic acid (1.087), gallic acid (0.710) and quercetin (0.703) were found to be in the leaves of S. cumini L. The Rf value of caffeic acid was 0.04, gallic acid was 0.95 and quercetin was 0.40. The chromatogram visualization was done at 254nm and 366nm. The statistical results demonstrated that this method is reproducible, determined, and highly reliable. This is the first circulation detailing the quantification of the compound, providing an essential method for the drug standardization.