Isolation and Identification of Secondary Metabolite Compound

Extract Etil Acetate from the Leaves of Durian Durio zibethinus L.

Dyna Grace Romatua Aruan

1,6

, Tonel Barus

, Ginda Haro

and Partomuan Simanjuntak

4,5

Postgraduate Chemistry Study Program, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara,

Jl. Bioteknologi No. 1 Kampus USU, Medan, Indonesia

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

Jl. Bioteknologi No. 1, Medan 20155, Indonesia

Faculty of Pharmacy, Universitas Sumatera Utara

Puslit Bioteknologi-LIPI, Jalan Raya Bogor Km 46, Cibinong, 16911, Indonesia

Department Pharmacy, University of Pancasila, Jagakarsa, Jakarta, Indonesia

Faculty of Pharmacy and Health Sciences, University of Sari Mutiara Indonesia, Medan 20123, Indonesia

Keywords: Durio zibethinus, Malvaceae, Steroid, Indonesian Plant, Secondary Metabolite.

Abstract: The aim of this study was to isolate and identify secondary metabolite in ethyl acetate extract of durian

leaves (Durio zibethinus L.). Isolation and purification is carried out by maceration, partitioning (ethyl

acetate and water), fractionation (column chromatography) and preparative Thin Latyer Chromatography

(TLC). Interpretation of infrared (IR) spectra and phytochemical screening showed that the secondary

metabolite is a steroid compound.

1 INTRODUCTION

Indonesia is known as a tropical area and even the

largest for the world scale with a variety of plants

that are of high value to the community, namely

plants that have medicinal properties. One of them is

the durian plant. This plant is very familiar,

especially the fruit. Durian Leaves are traditionally

stew used as an antipyretic for children.

Phytochemicals of durian leaves (Durio zibethinus

L.) are flavonoids, steroids/terpenoids, and

glycosides in ethyl acetate extract (Aruan, 2019).

Steroids are a group of secondary metabolites. This

is very important in the medical field. Isolation in

the process of separating chemical contained in a

material includes four important stages of

maceration, partitioning, purification, and

identification. This research is based on the results

of Insanu 2011 study on flavonoids in ethanol

extracts from durian leaves. This study aim to

examine the chemical content of durian leaves, to be

developed further as a natural medicine.

2 MANUSCRIPT PREPARATION

2.1 Preparation Sample

Samples were collected from Tornaginjang Sibolga

Village, Medan, North Sumatra, Indonesia.

Fresh durian leaves are cleaned of dirt, weighed,

then dried in a drying cabinet that is not exposed to

direct sunlight, then the dried durian leaves are

crushed using a blender. Blending durian leaves

were tested for steroid / terpenoid identification. 1

gram of dried durian leaf is soaked with ethyl

acetate, then filtered using filter paper, the filtrate is

collected in a vaporizer cup and then evaporated on

a water bath and the Liebermann-Burchard reagent

is added through the wall of the evaporating cup. If

the color purple or red turns blue or blue or green

blue indicates the presence of steroids / terpenoids

(Harborne, 1987).

2.2 Pemurnian Sampel

Dried durian leaves, crushed using a blender,

macerated with ethanol, then partitioned using ethyl

acetate. Ethyl acetate extract was fractionated with

130

Aruan, D., Barus, T., Haro, G. and Simanjuntak, P.

Isolation and Identiﬁcation of Secondary Metabolite Compound Extract Etil Acetate from the Leaves of Durian Durio zibethinus L..

DOI: 10.5220/0008857801300132

In Proceedings of the 1st International Conference on Chemical Science and Technology Innovation (ICOCSTI 2019), pages 130-132

ISBN: 978-989-758-415-2

n-hexane: ethyl acetate (5: 1) eluent. The spot is

monitored under UV light at 265 and 365 nm and

then sprayed with 1% cerium sulfate in order to see

patches showing some amount of compound in the

ethyl acetate fraction. The results of the first column

chromatography obtained 8 fractions followed by

antidiabetic testing. Obtained F3 fraction which has

antidiabetic activity with 82.85% inhibition. F3

fraction was carried out by the second column

chromatography with eluent n-hexane: ethyl acetate

5: 1 and obtained fraction 6, 7, 8 with the same Rf

distance called F3-1 fraction. The F3-1 fraction was

then carried out preparative chromatography with

eluent n-hexane: ethyl acetate 5: 1 and obtained F3-

1-1 fraction with Rf 0.7 with a weight of 81 mg with

white powder. The purity fraction F3-1-1 was

determined by TLC and eluent n-hexane: ethyl

acetate 5: 1, n¬-hexane: acetone 5: 1, n-hexane-

chloroform 5: 1; 2-D TLC n-hexane: ethyl acetate 5:

1. Pure isolate F3-1-1 fraction was characterized and

identified using infrared spectrophotometry (IR).

3 RESULT AND DISCUSSION

The results of screening of phytochemical extract of

durian leaf ethyl acetate with Liebermann Burchad

reagent showed that the green solution of the

compound was a steroid group (Harborne, 1987).

The results of the identification of the F3-1-1

fraction using infrared spectrophotometry (IR) can

be seen in Figure 1 and the interpretation of the

spectrum in Table 1.

Figure 1: IR Spectrum F3-1-1.

In Table 1 shows the peak of the absorption

spectrum at 3410.15 cm-1 in the presence of OH

groups; at a wavelength of 2945.30 cm-1 there is a

stretching group C-H; at a wavelength of 1676.14

cm-1 group C = C alkene absorption peak; at the

other absorption peak 1454.33 cm-1 in the presence

of CH

groups; peak absorption spectrum of 1371.39

cm-1 indicates the presence of CH

groups; and peak

absorption spectrum at 1120.64 cm-1 indicates the

presence of C-OH groups (Cole, 1963). IR spectrum

information reinforces data that there are no

aromatic functional groups but only alkene groups in

isolation.

Table 1: Spektrum Interpretation of IR dari F3-1-1.

Wavenumber

-1

Functional Groups

3410.15 -OH

2945.30 C-H

1676.14 C=C

1454.33 CH

1371.39 CH

1120.64 C-O

4 CONCLUSIONS

The best effluent for purification of ethyl acetate

extract from durian leaves (Durio zibethinus L.) by

using thin layer chromatography (TLC) is n-hexane:

ethyl acetate (5: 1) eluent. The type of secondary

metabolites obtained from the separation of ethyl

acetate extract from durian leaves by screening,

TLC, and IR information are steroid groups.

ACKNOWLEDGEMENTS

This research was supported by Grant Assistance for

Scientific Research from the Ministry of Research

Technology and Higher Education of the Republic

of Indonesia, Research Center for the Indonesian

Institute of Biotechnology Science, LIPI, Serpong,

Indonesia, and Research Center for the Indonesian

Institute of Biotechnology. Sciences, LIPI,

Cibinong, Indonesia.

REFERENCES

D. G. R. Aruan., T. Barus., G. Haro., P. Simanjuntak.,

2019. Phytochemical Screening and Antidiabetic

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Extract from Durian Leaves (Durio zibethinus L.).

Oriental Journal of Chemistry, 2019, 35(1), 487-490

D. G. R. Aruan, T. Barus, G. Haro, P. Simanjuntak. 2019.

Toxicity and Antioxidant Activities of Extracts of N-

Hexane, H

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Isolation and Identiﬁcation of Secondary Metabolite Compound Extract Etil Acetate from the Leaves of Durian Durio zibethinus L.

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Insanu, M., 2011. Isolation Flavonoid from Durian Leaves

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