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Abstract
Piper betle L. plant or “Sirih” is widely found in Southeast Asia and its leaves are commonly eaten together with lime and arecanut. The major active compounds of P. betle leaves are polyphenols, for example, allylpyrocatechol (APC), that are reported to possess anti-inflammatory, antioxidant, and antimicrobial properties. We developed and validated a method for quantitative determination of APC in the extracts from betel leaves using high-performance liquid chromatography (HPLC). Extraction of APC was performed using Agilent 1100 series HPLC on a ZORBAX Eclipse column (C18 with 250 mm × 4.6 mm) with acetonitrile—0.01 M phosphoric acid as the mobile phase under gradient mode. The flow rate was 1.0 mL/min, and APC was detected using a UV detector (222 nm). Concentration of APC in the extract was determined from the peak areas by referencing them against a calibration curve developed from commercial analytical APC standard. The yield of APC from this HPLC extraction was 13.82% compared to the isolation of APC using silica gel column chromatography in which the yield of APC eluted was only 0.9% (w/w) of the extract (4.7 g). APC fraction isolated from this study will be used for the molecular analysis of its effect on regulation of oxidative stress response enzymes in Staphylococcus aureus. In addition, this study potentially helps in choosing the suitable extraction technique and developed HPLC method that can be used for routine quality control and standardized formulations of APC in P. betle leaf extract.
Acknowledgment
The authors acknowledge the technical assistance provided by the Analytical Unit, Faculty of Pharmacy, UiTM, in carrying out HPLC analysis.
Notes
Working range: 0.01–0.2 mg/mL, r 2: Correlation coefficient.
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