Flavonoid Compound from Rambutan Bark

(Nephelium lappaceum L.)

Helmina Br Sembiring

and Mesy Jelisa

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia

Keywords: Isolation, Nephelium lappaceum L., Flavonoids, Flavonol.

Abstract: Flavonoids included polyphenol compounds that are found in many plants which are important compounds

in human food.The flavonoid group, including flavanone, flavone, dihydroflavonol, catechin, flavonol,

flavan-3-ols, isoflavones, auron, anthocyanidins, proanthocyanidins and chalcones, has a general structure of

C6-C3-C6. The aims of this study is to isolate and identify the group of flavonoid compound contained in

rambutanbark (Nephelium lappaceum L.). Rambutan bark powder was extracted maceration with methanol.

Methanol extract was dissolved with ethyl acetate repeatedly until the solution was negative flavonoids. Ethyl

acetate extract was dissolved with methanol and partitioned with n-hexane. The methanol extract which is a

total of flavonoids separated by column chromatography using chloroform: methanol (90:10; 80:20 and 70:30

(v/v)). The isolates obtained were purified by preparative thin layer chromatography and producing flavonoid

glycoside, yellow amorphous with Rf value of 0.25 using the chloroform: ethyl acetate eluent (50:50) v/v.

The pure isolate obtained were analyzed by UV-Visible Spectropometer, FT-IR, and H-NMR. Based on the

interpretation of spectroscopic data, the flavonoid compound isolated from rambutan bark wasflavonol group.

1 INTRODUCTION

Rambutan (Nephelium lappaceum L.) is an evergreen

tree (Sukmandari et al., 2017), a plant that identical

with Southeast Asian countries, in some areas of

Indonesia (Wahini et al., 2018). The dried rambutan

fruit peel is used in traditional medicine, cooking and

in the manufacture of soap. The roots bark and

rambutan leaves have various uses in medicine and in

the production of dye (Suganthi & Josephine, 2016).

Rambutan fruit peel contains flavonoids, tannins and

saponins (Hariana, 2006).

Flavonoids are the most extensive groups of

phenolics Flavonoids are secondary with low

molecular weight that have bioactivity (Weston &

Mathesiu, 2013). Flavonoids are polyphenol

compounds composed of 15 carbon atoms, with two

aromatic rings connected by a bridge consisting of

three carbon atoms (Crozier et al., 2006). The

flavonoid group, including flavanone, flavone,

dihydroflavonol, catechin, flavonol, flavan-3-ols,

isoflavones, auron, anthocyanidins,

proanthocyanidins and chalcones, has a general

structure of C6-C3-C6 (Rosa et al., 2010).

Flavonoids have a positive effect on human and

animal health. Flavonoids are widely used as a

therapy for disease and chemoprevention (Panche et

al., 2016). Bioactivities of these phenolic,

polyphenolic acids or essential oil are potential as

new leads for the development of pharmaceutical

(Saranya et al., 2017), antibacterial (Sembiring et al.,

2019 )and agricultural products to improve human

health and nutrition (Khadem & Marles, 2010)

.Flavonoids are able to treat diseases, such as cancer

and heart disease (Zhang et al., 2015) can be used to

protect the human body from free radicals (Megawati

et al., 2015), (Molyneux, 2004) and can reduce the

risk of cancer and inflammation (Kumar & Pandey,

2013).

Pangalinan et al., (2012) mentioned that rambutan

bark are effective as antifungal against Candida

albicans. The purpose of this study was to isolated

and identified the flavonoid group contained in the

bark of the rambutan (N. Lappaceum L.). Flavonoid

compounds were isolated by maceration extraction

and column chromatography methods. Flavonoid

compound identification was carried out by FT-IT,

UV-Visible and HNMR spectroscopy.

Br Sembiring, H. and Jelisa, M.

Flavonoid Compound from Rambutan Bark (Nephelium lappaceum L.).

DOI: 10.5220/0010139100002775

In Proceedings of the 1st International MIPAnet Conference on Science and Mathematics (IMC-SciMath 2019), pages 205-208

ISBN: 978-989-758-556-2

205

2 METHODS

2.1 Material

Rambutan bark was obtained from Gelugur Rimbun

Village, Pancur Batu District, Deli Serdang, North

Sumatra, Indonesia. Identification of plant was done

at Herbarium Medanense (MEDA) Universitas

Sumatera Utara. All chemicals used such as Silica Gel

(70 – 230 mesh), for column chromatography, FeCl

NaOH, Serbuk Mg, HCl

(p)

, H

4(p)

, kloroform,

silika gel 60 F

254

for thin layer chromatography, KLT

Preparative 60 F

254

and methanol were from E Merck,

methanol and ethyl acetate as solvent were distilled

before used (Saldanha et al., 2013).

2.2 Instrument

The

H NMR spectra were recorded with a (Agilent

500 MHz, Frekuensi 500 MHz), spectrometer

instrument with CD

OD as a solvent and TMS as an

internal standard and chemicalshifts are given in δ

(ppm). IR spectra were recorded on FT-IR (type

Mb3000, 485 – 8500 cm

-1

), UV spectra were recorded

on Spektrofotometer UV-Visible (Type UV – 1800

Shimadzu, 190 – 1100 nm), evaporation of solvents

with rotary evaporator (Heidolph), spotting

monitoring with lights of UV(254nm/356nm, UVGL

58).

2.3 Procedure

Isolation flavonoid compounds were done based on

(Megawati et al., 2015) and (Hostettmann et al.,

1995) with a slight modification. Rambutan bark

powder (1900 g) was macerated for 2 days using 11

L methanol (until all samples were submerged with

methanol). Maserat is accommodated and the solvent

is evaporated with a rotary evaporator and dried with

a water bath until a dry methanol extract was

obtained. Dry methanol extract was re-extracted with

ethyl acetate to separate tannins. The filtrate obtained

was evaporated with a rotary evaporator and water

bath until all the ethyl acetate solvent evaporated.

Ethyl acetate extract was redissolved with methanol

and repartitioned with n-hexane until the n-hexane

layer was colorless. The methanol layer was re-

concentrated with a rotary evaporator and dried with

a water bath to obtain a dry extract of methanol.

The dried methanol extract (5g) was added to the

column chromatographic containing silica gel slurry,

eluted with chloroform: methanol (90:10; 80:20;

70:30 v / v) slowly. The isolates were collected in

vials every 10 ml, then analyzed with TLC. The

fractions that have the same Rf value are combined.

Fractions of 41-85 have the same Rf value, combined

then purified with preparative TLC with cloroform:

etil asetat (40:60) (v/v), ) and produced one band spot

at the Rf 0.25. The band spot was crushed, eluted with

metanol: etil asetat (1:1)

, evaporated to obtain 7.9

mg pure isolate in the form of yellow amorphous. The

pure isolate was identification by UV-Vis, FT-IR and

H-NMR spectroscopy.

3 RESULTS AND DISCUSSION

The sample used in this study was the rambutan bark,

Nephelium lappaceum L. (Figure 1A), family

Sapindaceae. The pure isolate isolated from the bark

rambutanis a yellow amorphous (Figure 1B) and

identified by using UV-Vis, FT-IR and

H-NMR

spectroscopic analysis.

Figure 1: A. Rambutan plant; B. isolate.

The UV-Visible (CH

OH) spectra was shown in

Figure 2. Based on the spectra, the isolate isolated

from rambutan bark was fllavonol group, because

presence of spectraat λ

max

280,00 nm. Absorption

band II at maximum wavelength (max λ) 280.00 nm

is a flavonoid of the flavonol group (Andersen &

Markhan, 2006). FT-IR spectra of pure isolated was

shown in Figure 3. The FT-IR spectra for pure

isolates showed (KBr, ν max, cm

-1

) 3433.29(O-H),

2854,65 and 2924,09 (C-H sp

stretching),1627.92

(C=O),1527.62(C=C), 1381.03 (C-Hsp

bending) and

1273,02(C-O).All of these vibrations are common

vibrations found in flavonoid compounds. The

stretching vibration of the C-H sp

bond is not

detected because it overlaps with the broad vibration

of the O-H bond (Pavia et al., 2001).

IMC-SciMath 2019 - The International MIPAnet Conference on Science and Mathematics (IMC-SciMath)

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

RambutanBark

Figure 2: The UV-Visiblespectra of isolate.

Figure 3: The FT-IR Spectra of pure isolate.

Figure 4: The

H NMR spectra of pure isolate.

H NMR spectra of pure isolate wasdepected in

Figure 4. Based on 1H NMR spectra (Methanol-D6,

500 MHz, (ppm)), chemical shift (δ (ppm)): δ 5.914

and 5.918 (1H, d, H-6), δ 5.939 and 5.944 (1H, d, H-

8), δ 6.750 and 6.766 (1H, d, H-5’), δ 6.790 and 6.807

(1H, d, H-6’) and δ 6.972 and δ 6.977 (1H, d, H-2’),

)), the isolate had five aromatic protons and eight

rhamnose protons. The type and number of protons in

this isolate are the same as the type and number of

protons of quercetin compounds reported by Huang

et al.,(2013) But it has a slight difference in chemical

shift and splitting pattern, due to differences in the

frequency of the spectroscopic used. Substituent -OH

bind toC-3, C-7, C-3’, dan C-4’. However, C-5 has a

glycoside bond because no -OH peaks are found at a

chemical shift of 12.22 ppm (Claramunt et al., 2006).

Rhamnose is thought to bind to flavonoids because of

the presence of peaks δ (ppm) 4,1702 dan δ 4,1792

(1H, d, H-1’’), δ 3,6495 (4H, m, H-2’’,H-3’’,H4’’,H-

5’’) and δ 1.289 and 1.307 (3H, d, -

rhamnoside)ofrhamnose. The chemical shift of

rhamnose protons are similar to the chemical shift of

rhamnoseprotons reported by Plazonić et al, 2009)

Based on interpretation data of the UV-Visible,

FT-IR and 1H-NMR spectra, the flavonoid compound

isolated from rambutan bark was flavonol group

bound torhamnose at C-5with the structure shown as

in Figure5.

Figure 5: Structure of isolate.

4 CONCLUSIONS

Isolate obtained from 1900 g rambutan bark (N.

lappaceum L.) is a flavonoid 7.9 mg yellow

amorphous with an Rf value of 0.25 using the

chloroform: ethyl acetate eluent (50:50) v / v. Based

on the spectra and interpretation data the flavonoid

compound isolated from rambutan bark was flavonol

group.

Flavonoid Compound from Rambutan Bark (Nephelium lappaceum L.)

207

ACKNOWLEDGEMENTS

We would like to thank to Herbarium medananse

(MEDA), Laboratory of Natural Sciences Chemistry

Faculty of mathematical and Science Universitas

Sumatera Utara. We would also like to thank to Roch

Fitni for analysis of Spektrofotometer UV Visible and

FT-IR LPPT UGM Jl. Kaliurang Km. 4 Sekip Utara

Yogyakarta andand Elvira Hermawati for the analysis

H-NMR, Laboratory of Organic Chemistry, ITB

Bandung.

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