Isolation and Identification of DPPH Radical (2,2-diphenyl-1-pikrylhidrazyl) Scavenging Active Compound in Ethyl acetat fraction of Piper acre Blume

https://doi.org/10.22146/mot.48173

Hifdzur Rashif Rijai(1*), Nanang Fakhrudin(2), Subagus Wahyuono(3)

(1) Faculty of Pharmacy, Universitas Mulawarman, Samarinda, East Kalimantan, Indonesia
(2) Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia
(3) Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia
(*) Corresponding Author

Abstract


Piper acre Blume, known as Black Betel (local name), is a plant that is widely used by the people of East Kalimantan, especially in Samarinda, for the treatment of illness. Leaves (3-4 months old) are collected from Samarinda, extracted, fractionated, and monitored by DPPH antiradical activity. The isolation of the Piper acre Blume is performed on the active fraction, and the structure identification is based on spectroscopic data of the compound.  The leaves were dried, pulverized, and macerated with MeOH. Dried MeOH extract was obtained upon evaporation of the solvent. The extract was then fractionated by vacuum liquid chromatography (vlc), eluted gradually by solvents having different polarities (n-hexane, ethyl acetate and methanol). The fractions obtained were monitored using TLC [n-hexane: ethyl acetate (3: 1 v/v)] that was visualized by UV254 nm, UV366 nm and DPPH. The isolation was performed by preparative TLC [SiO2, n-hexane: ethyl acetate (3: 1)] on ethyl acetate fraction that showed the highest DPPH antiradical value. A single compound was obtained, and it appeared as a round spot and pure according to TLC performances at 3 different solvent systems. The isolated Piper acre Blume compound displayed the IC50 value on the anti-radical DPPH (measured at λ 520 nm) as 10.41µg/mL. The IR spectrum (KBr) showed –OH band (3450 cm-1), aliphatic bands [alkene, 3010 cm-1; alkana 2900 cm-1), an aromatic overtone bands (1900-200 cm-1) and a strong C=O band (1725 cm-1). The NMR (1H- and 13C-) (mono and 2D) indicated the present of a p-di-substituted aromatic signals (δ, 7.54 and 7.52, d, J =6 Hz, 1 H each), 2 methyl (δ, 0.96, d, J = 7.0 Hz, 6 Hs), a triplet signal (δ, 4.22 ppm). Other signals of CH- and CH2 were shown as m signals at δ, 1.64 and 1.34 ppm.  Based on those data, the compound was identified as isoamyl p-OH benzoate that is grouped as parabens used as a preservative in the pharmaceutical preparations. In conclusion, the anti-radical (DPPH) active compound present in the leaves of Piper acre Blume is identified as isoamyl p-OH benzoate, having IC50 value anti-radical DPPH 10,41µg/mL.


Keywords


anti-radical; DPPH; ethyl acetate; isoamyl p-OH benzoate; Piper acre Blume

Full Text:

PDF


References

Fessenden, R.J. and Fessenden, J.S., 1994. Organic Chemistry, Third. ed. Erlangga.

Molyneux, P., 2004. The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin J. Sci. Technol, , 2 26: 211–219.

Rija’i, A.J., 2013. ' Phytochemical Study of Secondary Metabolite Content in Piper acre Blume. Extracts and Bioactivity Tests on Shrimp Larvae (Artemia Salina Leach)', Skripsi, Islamic Bandung University, Bandung.

Rija’i, H.R., Syafnir, L., dan Rismawati, E., 2015. Antioxidant Activity Test of Black Betel Leaf Extract (Piper acre Blume.) With DPPH Free Radical Reduction (1,1-difenil-2-pikrilhidrazil). Prosiding SPeSIA, 58–64.

Rija’i, L., Kuncoro, H., dan Herman, 2015. Forest betel (P. acre Blume) a Wild Plant from East Kalimantan, Indonesia Potentially as Resources of Drug Compounds for Cancer. Journal of Pharmaceutical Sciences and Research, 7: 401–404.

Wahdaningsih, S., Prawita Setyowati, E., dan Wahyuono, S., 2011. Free Radical Scavenging Activity of Alsophila glauca J. Sm. Traditional Medicine Journal, 16: 156 – 160.




DOI: https://doi.org/10.22146/mot.48173

Article Metrics

Abstract views : 3084 | views : 3137

Refbacks





Copyright (c) 2019 Majalah Obat Tradisional

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

©Majalah Obat Tradisional (Traditional Medicine Journal)
 ISSN 2406-9086
Faculty of Pharmacy
Universitas Gadjah Mada