Development of a Simple High-Performance Liquid Chromatography-Based Method to Quantify Synergistic Compounds and Their Composition in Dried Leaf Extracts of Piper Sarmentosum Roxb.

There is a growing demand to enhance pharmaceutical and food safety using synergistic compounds from Piper sarmentosum Roxb., such as polyphenols and water-soluble vitamins. However, information on standardized analytical methods to identify and quantify these compounds of interest is limited. A reversed-phase high-performance liquid chromatography with diode-array detection (HPLC-DAD)-based method was developed to simultaneously detect and quantify the amounts of tannin, flavonoid, cinnamic acid, essential oil, and vitamins extracted from P. sarmentosum leaves using methanol, chloroform, and hexane. Commercially and non-commercially-cultivated P. sarmentosum leaves were subjected to seven different drying treatments (shade; sun; air oven at 40 °C, 60 °C, 80 °C, and 100 °C; and freeze-drying) for three consecutive months. Most compounds were detected most efficiently at a detection wavelength of 272 nm. The developed method displayed good detection limits (LOD, 0.026–0.789 µg/mL; LOQ, 0.078–2.392 µg/mL), linearity (R2 > 0.999), precision (%RSD, <1.00), and excellent accuracy (96–102%). All P. sarmentosum leaf extracts were simultaneously tested and analytically compared without time-consuming fractionation. Methanolic plant extracts showed better peak area and retention time splits compared to chloroformic and hexanoic extracts. Differences in synergistic compound composition were dependent on the type of drying treatment but not on cultivation site and time of sampling. Flavonoid was identified as the dominant phytochemical component in P. sarmentosum leaves, followed by the essential oil, cinnamic acid, ascorbic acid, and tannin. Overall, we present a simple and reproducible chromatographic method that can be applied to identify different plant compounds.