A reversed-phase HPLC-DAD was developed and validated to estimate the phenolic acids (gallic acid, caffeic acid, syringic acid, p-coumaric acid, sinapic acid, and ferrulic acid), flavonoids (catechin rutin, myricetin, quercetin, apigenin, and kaempferol), ascorbic acid, and eugenol. The chromatogram condition was set in suitable wavelengths 272 nm and run flow rate 0.7 µl/min using HPLC Agilent Technologies 1260 Infinity, a reversed-phase Zorbax SB-C18 column (3.5 µm particle size, i.d. 4.6 x 250 mm) with mobile phase solution (1:9, HPLC-grade acetonitrile:1% acetic acid). The linearity, precision, LOD, LOQ, and accuracy were R2>0.9907, RSD
Key words: Phenolic compounds, flavonoids, essential oil, Piper betle L., gradient HPLC, different solvent extracts.
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