Isolation and Qualitative Test of Piperine from Black Pepper (Piper nigrum L.) Methanol Extract

Abstract

Black pepper (Piper nigrum L.) is a spice commodity rich in alkaloids, in the form of piperine which gives a spicy taste and has various pharmacological activities such as anti-inflammatory, antioxidant, and anticancer properties. This study aims to isolate and perform qualitative validation of piperine compounds from black pepper fruit methanol extract through a series of phytochemical stages. Soxhlet extraction using methanol solvent yielded a viscous extract with a 10.3% yield. Furthermore, piperine separation was carried out by adding a 10% KOH solution in methanol to neutralize fatty acid and resin impurities, allowing neutral piperine to be separated. The purification stage through crystallization, washed with cold methanol, resulted in an isolate in the form of yellow needle-shaped crystals with a yield of 0.22%. Through phytochemical screening, the crystal was found to contain alkaloids because it gave a positive reaction to Mayer, Dragendorff, and Bouchardat reagents. Thin Layer Chromatography (TLC) analysis showed a single spot with an Rf value of 0.74, in accordance with literature standards. More specific identification using UV-Vis spectrophotometry showed maximum absorption at a wavelength of 342 nm, and melting point test results at 129.9°C matched literature values, confirming the purity of the piperine isolate. Therefore, the overall procedures proved effective for piperine isolation.