Abstract
This study isolated and identified the antimicrobial compounds of Philippine Piper betle L. leaf ethanol extracts by thin layer chromatography- (TLC-) bioautography and gas chromatography-mass spectrometry (GC-MS). Initially, TLC separation of the leaf ethanol extracts provided a maximum of eight compounds with R f values of 0.92, 0.86, 0.76, 0.53, 0.40, 0.25, 0.13, and 0.013, best visualized when inspected under UV 366 nm. Agar-overlay bioautography of the isolated compounds demonstrated two spots with R f values of 0.86 and 0.13 showing inhibitory activities against two Gram-positive multidrug-resistant (MDR) bacteria, namely, methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus. The compound with an R f value of 0.86 also possessed inhibitory activity against Gram-negative MDR bacteria, namely, carbapenem-resistant Enterobacteriaceae-Klebsiella pneumoniae and metallo-β-lactamase-producing Acinetobacter baumannii. GC-MS was performed to identify the semivolatile and volatile compounds present in the leaf ethanol extracts. Six compounds were identified, four of which are new compounds that have not been mentioned in the medical literature. The chemical compounds isolated include ethyl diazoacetate, tris(trifluoromethyl)phosphine, heptafluorobutyrate, 3-fluoro-2-propynenitrite, 4-(2-propenyl)phenol, and eugenol. The results of this study could lead to the development of novel therapeutic agents capable of dealing with specific diseases that either have weakened reaction or are currently not responsive to existing drugs.
Highlights
Piper betle L. belonging to the family Piperaceae is a climbing vine used in alternative medicine due to its numerous therapeutic properties, which include its antibacterial, antifungal, antioxidant, cytotoxic, antihelminthic, antiprotozoal, antidiabetic, hepatoprotective, and immunomodulatory properties [1]
This study aimed to identify and isolate the antibacterial compounds of P. betle extracts using Thin Layer Chromatography (TLC), agar-overlay, and contact bioautographic assays and to identify the semivolatile and volatile compounds using gas chromatography-mass spectrometry (GC-mass spectrometry (MS))
To isolate and identify the bioactive compounds of the P. betle leaf extract, TLC was initially performed as a qualitative method to document the extract constituents
Summary
Piper betle L. belonging to the family Piperaceae is a climbing vine used in alternative medicine due to its numerous therapeutic properties, which include its antibacterial, antifungal, antioxidant, cytotoxic, antihelminthic, antiprotozoal, antidiabetic, hepatoprotective, and immunomodulatory properties [1]. The plant is known to be widely distributed in India, Sri Lanka, Malaysia, Indonesia, Thailand, China, Philippines, and other subtropical countries [2]. It has been reported to have broad spectrum antimicrobial activities against various bacterial strains [1] and fungi [3, 4]
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