6/26/2020 About the Journal – Selamat Datang ke Laman Web MJAS

https://mjas.analis.com.my/index.php/about-the-journal/ 1/2

About this Journal

Malaysian Journal of Analytical Sciences (MJAS) is a peer-reviewed research journal

that is devoted to the dissemination of new and original knowledge in all branches of

analytical chemistry. Manuscripts submitted may be experimentally based or may be

entirely theoretical and should contribute to any phase of analytical measurements and

concepts including electrochemistry, mass spectrometry, environmental analysis,

separation techniques, spectroscopy, instrumentation, surface analysis, bioanalysis,

structures, chemometrics and data processing. Manuscripts dealing with conventional

analytical methods should offer a significant, original application of the method, a

noteworthy improvement or results on an important analyte.

Malaysian Journal of Analytical Sciences is indexed in MyAIS (Malaysian Abstracting

and Indexing System), International Nuclear Information System (INIS Database) and

SCOPUS (Yearly SJR 2018 0.18, SiteScore 0.6, Impact per Publication (0.375), Quartile

Q4 (Analytical Chemistry) and H-index of 9). MJAS is also cited in MyCite, an indexing

system owned by Malaysian Citation Centre (MCC), the Ministry of Higher Education

(MOHE) Malaysia, with a 5-year Impact Factor of 0.061 (2017), Cited half-life of 2.8,

Q2 (Sciences), Q4 (Analytical Sciences) and H-index of 3.

ABOUT THE JOURNAL

Selamat Datang ke Laman Web MJAS

HOME THE JOURNAL ADVISORS & EDITORIAL BOARD INFO LIST OF ISSUES

CONTACT IMPORTANT LINKS

6/26/2020 About the Journal – Selamat Datang ke Laman Web MJAS

https://mjas.analis.com.my/index.php/about-the-journal/ 2/2

Q2 (Sciences), Q4 (Analytical Sciences) and H index of 3.

This Journal is published by the Malaysian Analytical Sciences Society (ANALIS) as

the sole scientific publication of the Society since 1995. From 2015, the Journal is

published in six issues per volume (per year) i.e. in February, April, June, August,

October and December.

Malaysian Journal of Analytical Sciences provides immediate open access to its content

on the principle that making published articles freely access and downloadable to the

public supports a greater global exchange of knowledge. Researchers from any countries

are invited to publish their research articles in this journal.

 

Publication Policy

Peer Review Process

Guidelines to Authors

MJAS Template Format

Malaysian Journal of Analytical Sciences

6/26/2020 Advisors & Editorial Board – Selamat Datang ke Laman Web MJAS

https://mjas.analis.com.my/index.php/advisors-editorial-board/ 1/4

International Advisors

Members

Fortunato Servilla III

(Philippines)

Muhammad Idiris Saleh

(Malaysia)

Ganden Supriyanto

(Indonesia)

R. Narayanaswamy

(U. Kingdom)

ADVISORS & EDITORIAL BOARD

Selamat Datang ke Laman Web MJAS

HOME THE JOURNAL ADVISORS & EDITORIAL BOARD INFO LIST OF ISSUES

CONTACT IMPORTANT LINKS

6/26/2020 Advisors & Editorial Board – Selamat Datang ke Laman Web MJAS

https://mjas.analis.com.my/index.php/advisors-editorial-board/ 2/4

Hassan Y. Aboul-Enein

(Egypt)

Sukiman Sarmani

(Malaysia)

Lee Hian Kee

(Singapore)

Wan Md Zin Wan Yunus

(Malaysia)

Editorial Board 2020

Editor-in-Chief

Md Pauzi Abdullah

(Universiti Kebangsaan Malaysia)

Editors

Ahmad Saat

(Universiti Teknologi MARA)

Amran Ab. Majid

(Universiti Kebangsaan Malaysia)

Bahruddin Saad

(Universiti Teknologi PETRONAS)

6/26/2020 Advisors & Editorial Board – Selamat Datang ke Laman Web MJAS

https://mjas.analis.com.my/index.php/advisors-editorial-board/ 3/4

Lee Yook Heng

(Universiti Kebangsaan Malaysia)

Mansor Ahmad

(Universiti Putra Malaysia)

Mhd Radzi Abas

(Universiti Malaysia Perlis)

Mohd Marsin Sanagi

(Universiti Teknologi Malaysia)

Mohd Talib Latif

(Universiti Kebangsaan Malaysia)

Nor Azah Yusof

(Universiti Putra Malaysia)

Norhayati Mohd Tahir

(Universiti Malaysia Terengganu)

Suhaimi Suratman

(Universiti Malaysia Terengganu)

Wan Aini Wan Ibrahim

(Universiti Teknologi Malaysia)

Zani @ Zaini Assim

(Universiti Malaysia Sarawak)

Wan Mohd A�q Wan Mohd Khalik

(Universiti Malaysia Terengganu)

Executive Editor

Harun Haji Hamzah

Site Administrator

6/26/2020 Malaysian Journal of Analytical Sciences (MJAS)

ejournals.ukm.my/mjas/index 1/1

OPEN JOURNAL SYSTEMS

Journal Help

USER

Username

Password ••••••••

Remember meLogin

NOTIFICATIONS

ViewSubscribe

FONT SIZE

INFORMATION

For ReadersFor AuthorsFor Librarians

HOME ABOUT LOGIN REGISTER ANNOUNCEMENTS ETHICS STATEMENT USER

GUIDES CURRENT ISSUES

Home > Malaysian Journal of Analytical Sciences (MJAS)

Malaysian Journal of Analytical Sciences (MJAS)

Malaysian Journal of Analytical Sciences (MJAS) is a peer-reviewed research journal that is devoted to thedissemination of new and original knowledge in all branches of analytical chemistry. Manuscripts submittedmay be experimentally based or may be entirely theoretical and should contribute to any phase of analyticalmeasurements and concepts including electrochemistry, mass spectrometry, environmental analysis,separation techniques, spectroscopy, instrumentation, surface analysis, bioanalysis, structures,chemometrics and data processing. Manuscripts dealing with conventional analytical methods should offer asignificant, original application of the method, a noteworthy improvement or results on an importantanalyte.

Malaysian Journal of Analytical Sciences is indexed in MyAIS (Malaysian Abstracting and Indexing System),International Nuclear Information System (INIS Database) and SCOPUS. MJAS is also cited in MyCite, anindexing system owned by Malaysian Citation Centre (MCC), the Ministry of Higher Education (MOHE)Malaysia, with a 5-year Impact Factor of 0.035 (2011), Cited half-life of 2.3 and H-index of 2.

This Journal is published by the Malaysian Analytical Sciences Society or ANALIS as the sole scientificpublication of the Society since 1995. From this year (2015), the Journal is published in six issues pervolume (per year) i.e. in February, April, June, August, October and December.

Malaysian Journal of Analytical Sciences provides immediate open access to its content on the principle thatmaking published articles freely access and downloadable to the public supports a greater global exchangeof knowledge. Researchers from any countries are invited to publish their research articles in this journal.

ISSN: 1394-2506

made_pharmawa

6/26/2020 Malaysian Journal of Analytical Sciences - Impact Factor, Overall Ranking, Rating, h-index, Impact Factor, SJR, Publisher, ISSN, Scientifi…

https://www.resurchify.com/all_ranking_details_2.php?id=19171 1/6

SEARCH

Malaysian Journal of Analytical Sciences - Impact Factor, Overall Ranking, h-index, SJR, Rating, Publisher, ISSN, and other Important Metrics

Title Malaysian Journal of Analytical Sciences

Publication Type journal

Subject Area, Categories,Scope

Analytical Chemistry (Q4)

h-index 9

Overall Rank/Ranking 19171

SCImago Journal Rank(SJR)

0.18

Impact Factor 0.53

Publisher Faculty of Science and Technology, Universiti Kebangsaan Malaysia

Country Malaysia

ISSN 13942506

Enter Journal Title, Conference Name, Book Title or Publisher Name

2020 call for papersPeer Reviewed Journal, Indexed, Fast.

asrjetsjournal.org OPEN

resurchify

6/26/2020 MJAS | Malaysian Journal of Analytical Sciences

https://mjas.analis.com.my/mjas/v24_n1/v24_n1.html 1/2

Malaysian Journal of Analytical Sciences Vol 24 No 1 2020

Page

1. PREPARATION AND CHARACTERISATION OF pH PROBE MEMBRANE FOR DETERMINATION OF TOTAL ALKALINITY IN SEAWATER (Penyediaan dan Pencirian Membran Prob pH bagi Penentuan Jumlah Kealkalian dalam Air Laut) Noor Sheryna Jusoh, Azrilawani Ahmad, Marinah Mohd Ariffin, Hafiza Mohamed Zuki

| Abstract & References || Full Text: pdf ( 1131 K ) |

1

2. SYNTHESIS AND SOLID-STATE STRUCTURAL ELUCIDATION OF RHENIUM(I) COMPLEX WITH 1-CINNAMOYL-3-(PYRIDIN-2-

YL)PYRAZOLE(Sintesis dan Penentuan Struktur Pepejal Kompleks Renium(I) dengan 1-Sinamoil-3- (Piridin-2-il)Pirazola) Wun Fui Mark-Lee, Yan Yi Chong, Azizul Hakim Lahuri, Mohammad B. Kassim

| Abstract & References || Full Text: pdf ( 957 K ) |

11

3. STUDIES AND ABSORPTION ISOTHERMAL OF METHYLENE BLUE BY USING N,O-CARBOXYMETHYL CHITOSAN

(Kajian Kinetik dan Isoterma Serapan Metilena Biru Menggunakan N,O-Karboksimetil Kitosan) Putri Amirah Solehin Sulizi and Nadhratun Naiim Mobarak

| Abstract & References || Full Text: pdf ( 605 K ) |

21

4. AGAROSE-CHITOSAN-INTERGRATED MULTIWALLED CARBON NANOTUBES FILM SOLID PHASE MICROEXTRACTION COMBINED

WITH HIGH PERFORMANCE LIQUID CHROMATOGRAPHY FOR THE DETERMINATION OF TRICYCLIC ANTIDEPRESSANT DRUGS INAQUEOUS SAMPLES (Filem Agarosa-Kitosan Bersepadu Nanotiub Kabon Berbilang Dinding Pengestrakan Mikro Fasa Pepejal Digabungkan DenganKromatografi Cecair Prestasi Tinggi-Pengesanan Ultralembayung Untuk Penentuan Anti-Murung Trisiklik Di Dalam SampelAkueus) Wan Nazihah Wan Ibrahim, Mohd Marsin Sanagi, Nor Suhaila Mohammad Hanapi, Nursyamsyila Mat Hadzir, Noorfatimah Yahaya,Sazlinda Kamaruzaman

| Abstract & References || Full Text: pdf ( 747 K ) |

33

5. APPLICATION OF BOX-BEHNKEN DESIGN WITH RESPONSE SURFACE METHODOLOGY FOR OPTIMIZING OXYGEN COLOUR

INDICATOR FOR ACTIVE PACKAGING(Aplikasi Reka Bentuk Box-Behnken dengan Kaedah Gerak Balas Permukaan untuk Mengoptimumkan Penunjuk OksigenBerwarna bagi Pembungkus Aktif) Aishah Mohd Marsin and Ida Idayu Muhamad

| Abstract & References || Full Text: pdf ( 1405 K ) |

42

6. APPLICATION OF MICROWAVE-ASSISTED EXTRACTION COUPLED WITH DISPERSIVE LIQUID-LIQUID MICROEXTRACTION FOR

THE DETERMINATION OF POLYCYCLIC AROMATIC HYDROCARBONS IN VEGETABLES (Aplikasi Pengekstrakan Berbantukan Mikrogelombang Gandingan dengan Pengekstrakan Mikro Cecair-Cecair Serakan bagiPenentukan Hidrokarbon Aromatik Polisiklik dalam Sayur-Sayuran) Chai Mee Kin, Tan Yeong Hwang, Wong Ling Shing

| Abstract & References || Full Text: pdf ( 457 K ) |

53

7. CORROSION INHIBITION STUDY ON GLYCEROL AS SIMULTANEOUS GAS HYDRATE AND CORROSION INHIBITOR IN GAS

PIPELINES (Kajian Perencatan Kakisan oleh Gliserol sebagai Perencat Serentak untuk Hidrat Gas dan Kakisan dalam Saluran Paip Gas) Vinayagam Sivabalan, Belkhir Walid, Yoann Madec, Ali Qasim, Bhajan Lal

| Abstract & References || Full Text: pdf ( 803 K ) |

62

8. PHYTOCHEMICAL SCREENING AND FTIR SPECTROSCOPY ON CRUDE EXTRACT FROM Enhalus acoroides LEAVES

(Saringan Fitokimia dan Spektroskopi FTIR Ekstrak Mentah Daun Enhalus acoroides) Made Pharmawati and Luh Putu Wrasiati

| Abstract & References || Full Text: pdf ( 554 K ) |

70

9. EFFECT OF BMIM-CHLORIDE ON PHYSIO CHEMICAL PROPERTIES OF NANOFIBER MEMBRANE FOR DOMESTIC WASTEWATER

TREATMENT (Kesan BMIM-Klorida ke atas Sifat-Sifat Fisio-Kimia Membran Nanogentian untuk Rawatan Sisa Air Domestik) Ahmad Tarmizi Mohd, Nur Syakinah Abd Halim, Mohd Dzul Hakim Wirzal, Muhammad Roil Bilad, Nik Abdul Hadi Md Nordin,Zulfan Adi Putra, Abdull Rahim Mohd Yusoff

| Abstract & References || Full Text: pdf ( 779 K ) |

78

10. CHEMICAL COMPOSITION OF ESSENTIAL OILS FROM LEAF EXTRACT OF PANDAN, Pandanus amaryllifolius ROXB.

(Komposisi Kimia Minyak Pati daripada Ekstrak Daun Pandan, Pandanus amaryllifolius Roxb.) 87

6/26/2020 MJAS | Malaysian Journal of Analytical Sciences

https://mjas.analis.com.my/mjas/v24_n1/v24_n1.html 2/2

Maisarah Mohamed Zakaria, Uswatun Hasanah Zaidan, Suhaili Shamsi, Siti Salwa Abd Gani

| Abstract & References || Full Text: pdf ( 674 K ) |

11. ELECTROCHEMICAL DETERMINATION OF DOPAMINE AND URIC ACID IN BLOOD SERUM USING ANIONIC SURFACTANTS ATCARBON PASTE ELECTRODES(Penentuan Dopamin dan Asid Urik dalam Serum Darah Secara Elektrokimia Menggunakan Surfaktan Anionik pada PerekatElektrod Karbon) Liridon Berisha, Egzontina Shabani, Arsim Maloku, Granit Jashari, Tahir Arbneshi

| Abstract & References || Full Text: pdf ( 1211 K ) |

97

12. MULTI-SPECTROSCOPIC AND CHEMOMETRICS ANALYSIS FOR FORENSIC DETERMINATION OF BLOOD SPECIES

(Multi-Spektroskopi dan Analisis Kemometrik untuk Penentuan Spesies Darah dalam Forensik) Durga Devi Sandran, Yusmazura Zakaria, Noor Zuhartini Md Muslim, Nik Fakhuruddin Nik Hassan

| Abstract & References || Full Text: pdf ( 594 K ) |

107

13. MAGNETIC KAOLINITE COMPOSITE FOR LEAD REMOVAL IN AQUEOUS SOLUTION

(Komposit Kaolinit Bermagnetik Untuk Penyingkiran Plumbum Dari Larutan Akueus) Izzan Salwana Izman, Siti Nor Atika Baharin, Ruhaida Rusmin

| Abstract & References || Full Text: pdf ( 1012 K ) |

115

14. PENENTUAN KEPEKATAN LOGAM BERAT DALAM SEDIMEN DI MUARA SUNGAI KEMAMAN, TERENGGANU

(Determination of Heavy Metal Concentrations in Sediment of Kemaman River Estuary, Terengganu) Ahmad Shamsuddin Ahmad, Suhaimi Suratman, Noor Azhar Mohamed Shazili, Ong Meng Chuan, Mohammed Faizal Abdul Rahim

| Abstract & References || Full Text: pdf ( 987 K ) |

125

15. INSIGHTS OF THE CHEMICAL CONSTITUENTS AND HYDROTHERMAL CARBONIZATION OF Crescentia cujete L.

(Pencirian Jujukan Kimia dan Pengkarbonan Hidrotermal bagi Crescentia cujete L.) Judith Clarisse Jose, Glenn Oyong, Michael Dominic Ajero, Irving Chiong, Esperanza Cabrera, Maria Carmen S. Tan

| Abstract & References || Full Text: pdf ( 1142 K ) |

134

16. EXTRACTION OF 4-OCTYLPHENOL AND 4-NONYLPHENOL IN RIVER WATER USING SOLID-PHASE EXTRACTION AND HIGH-

PERFORMANCE LIQUID CHROMATOGRAPHY(Pengekstrakan 4-Oktilfenol dan 4-Nonilfenol di dalam Air Sungai Menggunakan Pengekstrakan Fasa Pepejal dan KromatografiCecair Prestasi Tinggi) Mohd Zahid Baharom, Nurulnadia Mohd Yusoff, Wan Mohd Afiq Wan Mohd Khalik, Marinah Mohd Ariffin, Jamilah Karim, SyazrinSyima Sharifuddin

| Abstract & References || Full Text: pdf ( 776 K ) |

146

Malaysian Journal of Analytical Sciences, Vol 24 No 1 (2020): 70 - 77

70

S

PHYTOCHEMICAL SCREENING AND FTIR SPECTROSCOPY ON CRUDE

EXTRACT FROM Enhalus acoroides LEAVES

(Saringan Fitokimia dan Spektroskopi FTIR Ekstrak Mentah Daun Enhalus acoroides)

Made Pharmawati1* and Luh Putu Wrasiati2

1Biology Department, Faculty of Mathematics and Natural Sciences

2Department of Agro-Industrial Technology, Faculty of Agricultural Technology Udayana University, Kampus Bukit Jimbaran, Badung, 80361, Bali, Indonesia

*Corresponding author: made_pharmawati@unud.ac.id

Received: 25 November 2019; Accepted: 9 January 2020

Abstract Seagrass provides key ecological services in marine ecosystems, such as stabilising sediment, providing oxygen and acting as a nursery ground for marine biota. Seagrass has also been reported to have antioxidant activity that is useful for humans. One seagrass species, Enhalus acoroides, is widely distributed in Indonesia. This study aims to screen the phytochemical compounds,

determine the functional groups and evaluate the profile of pigments present in E. acoroides leaf extract, which were collected from Semawang Beach, Sanur, Bali, Indonesia. The leaf extract was prepared using chloroform: ethanol (9:1) and tested for the presence of saponin, phenols, tannins and flavonoids. The functional groups and pigment profile were determined via Fourier-transform infrared spectroscopy (FTIR) and thin-layer chromatography (TLC), respectively. The results showed that the E. acoroides leaf extract contained phenols, tannins and flavonoids. The major functional groups found in the leaf extract were hydroxyl groups, lipids, alkanes, secondary amines, fatty acids, benzenoid compounds and phenols. The FTIR analysis also identified the presence of chlorophyll and carotenoids in the extract, which was further supported by the TLC analysis. This research shows that E. acoroides is a potential source of antioxidants and provides an opportunity for the development of natural

products from E. acoroides in drug discovery. Keywords: Enhalus acoroides, FTIR, chromatography, phytochemical compound, seagrass

Abstrak Rumput laut memainkan peranan penting dalam ekosistem marin yang menstabilkan sedimen, membekalkan oksigen dan berfungsi sebagai halaman bagi biota marin. Rumput laut juga dilaporkan mengandungi aktiviti antioksidan yang bermanfaat bagi manusia. Spesis rumput laut, Enhalus acoroides di jumpai meluas di Indonesia. Kajian ini bertujuan menyaring sebatian fitokimia, penentuan kumpulan berfungsi dan menilai profil pigmen yang wujud di dalam ekstrak daun E. acoroides, yang

diambil dari Pantai Semawang, Sanur, Bali, Indonesia. Ekstrak daun disediakan menggunakan klorofom: etanol (9:1) dan diuji untuk penentuan kehadiran saponin, fenol, tannin dan flavonoid. Kumpulan berfungsi dan profil pigmen telah ditentukan masing-masing melalui spektroskopi inframerah transformasi Fourier (FTIR) dan kromatografi lapisan nipis (TLC). Hasil kajian menunjukkan ekstrak daun E. acoroides mengandungi fenol, tannin dan flavonoids. Kumpulan berfungsi utama yang dijumpai dalam ekstrak daun adalah kumpulan hidroksil, lipid, alkana, amina sekunder, asid lemak, sebatian benzoid dan fenol. Analisis FTIR juga mengenalpasti kehadiran klorofil dan karotenoid di dalam ekstrak, yang mana ia juga disokong oleh hasil analisis TLC. This research shows that E. acoroides is a potential source of antioxidants and provides an opportunity for the development of natural products from E. acoroides in drug discovery. Hasil kajian mendapati E. acoroides berpotensi sebagai sumber

antioksidan dan potensinya dalam pembangunan sumber semulajadi dalam penemuan ubat. Kata kunci: Enhalus acoroides, FTIR, kromatografi, sebatian fitokimia, rumput laut

Pharmawati & Wrasiati: PHYTOCHEMICAL SCREENING AND FTIR SPECTROSCOPY ON CRUDE

EXTRACT FROM Enhalus acoroides LEAVES

71

Introduction

Plant extracts have great potency and can be used for a variety of purposes. Approximately 80% of the world’s

population relies on traditional medicine for health care, and most therapies use plant extracts and their active

compounds [1], suggesting that two-thirds of all plant species have medicinal value [2]. Research has shown that

most medicinal plants contain antioxidant properties [3]. Nowadays, natural antioxidants are used in cosmetics,

foods and pharmaceutical products due to the antioxidants’ free radical scavenger activity. Exposure to environmental pollution, dietary xenobiotics and radiation leads to the generation of reactive oxygen species (ROS),

which induce oxidative stress [4]. Antioxidants prevent the formation of ROS, neutralise ROS and repair the

damage caused by ROS [5].

There are many plant species whose phytochemical content is unknown. One group of plants that has not yet been

explored in depth is seagrass, which is the only flowering plant (Angiospermae) that grows in a marine

environment. It can form a seagrass meadow that consists of one or a few dominant species [6]. In Indonesia, 13

seagrass species have been reported [7, 8], and nine species have been found in Bali alone [9]. One such species is

Enhalus acoroides (L.f.) Royle, the largest tropical seagrass species in Southeast Asian waters [10]. A previous

study on E. acoroides from India has reported that this species contains alkaloids, phenols, flavonoids, steroids,

tannins and saponin. Furthermore, it has been shown that aqueous extracts of E. acoroides have a high radical

scavenging activity [11]. Several studies on E. acoroides from Indonesia have shown that methanol extract of E. acoroides contains phenols, flavonoids and terpenes [12-14].

Several techniques are available to identify the phytochemical compounds in plant extracts. For example, Fourier-

transform infrared spectroscopy (FTIR) is a method used to identify the functional groups in gaseous, liquid and

solid materials via infrared radiation beams [15]. It has been used previously to identify the biomolecules present in

plant extracts from Erythrina variegata [16], Myristica dactyloides [17] and Urtica dioica [18] among others. Thin-

layer chromatography (TLC) is another method used to identify the compounds in a mixture via separating non-

volatile mixtures. Many previous studies have employed TLC to analyse the bioactive compounds in medicinal

plants [19, 20], plant pigments [21] and food additives and processing [22].

The objectives of this study were to screen the phytochemical compounds in the leaf extract of E. acoroides from Semawang Beach, Sanur, Bali, Indonesia, identify the pigments contained in the leaf extract and determine the

functional groups present in the E. acoroides leaves before and after extraction with chloroform: ethanol (9:1).

Materials and Methods

Preparation of extract

The E. acoroides leaves were collected from Semawang Beach, Sanur, Bali, Indonesia, washed and air-dried for

three days. The leaves were further dried in an oven at 50 ºC for one day until brittle. The dried leaves were

disrupted using a blender and sieved with a 60-mesh sieve. The voucher specimen was deposited in the herbarium

collection at Herbarium Biologi Udayana (HBU-MP10), Biology Department, Udayana University. The

identification of E. acoroides based on morphological characteristics was conducted following the processes of den

Hartog and Kuo [23] and McKenzie and Yoshida [24].

As much as 20 g of dried powder was extracted from 200 mL of chloroform: ethanol with a ratio of 9:1 (v/v). The

extraction was performed using a Soxhlet extractor for three hours. The solvent was filtered using Whatman filter

paper, and the filtrate was evaporated under a vacuum using a rotary evaporator at 40 ºC and 100 mbar (IKA®,

RV10). The extraction was performed three times.

Yield and phytochemical screening

The yield was expressed as a percentage and calculated by dividing the weight of the E. acoroides extract by the

weight of plant powder used and, then, multiplying by 100%. A preliminary phytochemical screening of the E.

acoroides extract was performed in triplicate using various chemical tests following standard methods [25] to

determine the presence of saponin, phenols, tannins and flavonoids. Saponin was tested via a foam test. As much as

2 ml of extract was added to water, and the mixture was shaken. The presence of saponin was indicated by the formation of foam. Phenols were determined by the ferric chloride test, in which ferric chloride was added to the

Malaysian Journal of Analytical Sciences, Vol 24 No 1 (2020): 70 - 77

72

E. acoroides extract. A positive result was shown by the change of colour from green to black. The presence of

tannins was tested by the addition of ferric chloride and sulphuric acid to diluted the extract, and a positive result

was shown by black colouring and the formation of black deposits. Flavonoids were tested by adding ammonia and

sulphuric acid to diluted extract; a yellow colour indicated the presence of flavonoids.

FTIR analysis The FTIR analysis was conducted on non-extracted leaf powder and the chloroform: ethanol-extracted sample. The

50 ºC oven-dried E. acoroides leaves were blended into a fine powder. As much as 1 mg of the sample was mixed

with 50 mg KBr (FTIR-grade); then, some of the mixture was placed into the sample holder. All investigations were

performed with an IRPrestige-21 (Shimadzu). The chloroform: ethanol-extracted E. acoroides leaf paste was diluted

with two drops of chloroform: ethanol (9:1). One drop of the diluted extract was then mixed with 50 mg KBr. The

scanning absorption range was 400 to 4000 cm−1.

TLC separation

The separation and identification of pigments were performed using TLC on a 20-cm x 20-cm cellulose plate

(Merck). The solvent mixture consisted of petroleum ether, acetone and n-propanol at a ratio of 90:10:0.45 [26].

The TLC cellulose plate was first activated by drying the plate in an oven at 105 °C for at least 45 minutes. The

extract was applied on the plate and allowed to dry. Then, the plate was placed in the TLC tank, and the spots were migrated.

Results and Discussion

The yield of the extract was 1.57% ± 0.23. The yield from this study, wherein chloroform: ethanol (9:1) was used,

was higher than the yields of previous E. acoroides extracts from Wakatobi, Sulawesi, Indonesia, and Madura, East

Java, Indonesia, wherein only chloroform was used as a solvent. The yield of the crude extract from Wakatobi was

0.55%, while that from Madura was 0.8% [27]. Chloroform is a non-polar solvent; therefore, it only extracts non-

polar compounds. Ethanol is a polar solvent, so the addition of ethanol in the present study caused the extraction of

more polar compounds in the leaf sample. Additionally, it is possible that the environmental growth conditions of E.

acoroides affected the extraction yield.

The phytochemical compounds contained in the extract of E. acoroides are shown in Table 1. Saponin was not

considered to be present in the extract because foam stability was not detected, i.e. the foam lasted less than five

minutes. However, phenols, tannins and flavonoids were detected in the chloroform: ethanol (9:1) E. acoroides leaf

extract. The presence of phenols and tannins in E. acoroides leaves from Indonesian coastal waters has been

previously reported as well [13, 27]. A previous methanol extract of E. acoroides leaves from East Lombok, Nusa

Tenggara Barat, Indonesia, was shown to contain phenols, tannins and terpenes, but flavonoids were not detected

[13]. Another study tested a methanol extract of E. acoroides leaves from the coastal waters of Ambon, Indonesia,

and found the presence of flavonoids [14]. Furthermore, a previous chloroform extract of E. acoroides leaves from

India was reported to contain flavonoids [11]. The presence of phenols, tannins and flavonoids in E. acoroides

indicates the seagrass species’ potential as a source of antioxidants.

Table 1. Qualitative phytochemical composition of chloroform: ethanol (9:1) leaf extract of E. acoroides

Compounds Colour Change Results

Saponin Unstable foam -

Phenols Dark green to black +

Tannins Dark green to black and black deposits formed +

Flavonoids Dark green to yellow ++

˗ = undetected, + = low, ++ = moderate

Pharmawati & Wrasiati: PHYTOCHEMICAL SCREENING AND FTIR SPECTROSCOPY ON CRUDE

EXTRACT FROM Enhalus acoroides LEAVES

73

The chemical bonds or functional groups present in the dried leaf powder and leaf extract of E. acoroides were

predicted using FTIR. The bonds were determined by interpreting the infrared absorption spectra. Figure 1 shows

the FTIR spectrum of the dried leaf powder, while Table 2 shows the interpretation of the chemical bonds in the

non-extracted leaf powder of E. acoroides. Strong bonds were found at 3552 cm−1, 1654.92 cm−1 and 2054 cm−1,

while the others varied from weak to medium. These results demonstrated the presence of hydroxyl groups, lipids,

alkanes, amino acids and phosphorus compounds.

Figure 1. FTIR spectrum of dried leaf powder of E. acoroides

Table 2. FTIR spectral peak values and functional groups of dried leaf powder of E. acoroides

Spectrum

No.

Wave Number

cm−1

Wave Number

cm−1

[28, 29]

Functional Group Assignment Predicted Compound

1 3552 3500–3650 v; O-H stretching vibration (free) Hydroxyl group

2 2945.30 ~2955 m; CH3 asymmetry stretching Lipid

3 2887.44 2880–2890 w; C-H stretching vibration Alkane

4 2360.87 2250–2700 m; NH component Amino acid, amino-related component

5 2054 2050–2140 m-w; symmetry -NH3+ stretching

vibration; broad

Free amino acid and their

hydrohalides

6 1654.92 1580–1660 s; C=C stretching vibration C=C conjugated with C=O

7 1411.89 1350–1410 w; C-H deformation vibration Secondary alcohol-free

8 1163.08 1155–1165 w-m; CH3 rocking vibration P-O-C2H5; Organic

phosphorus compound

9 927.76 925–930 m; C-C skeletal vibration Alkane

10 858.32 820–920 w-m; C-C skeletal vibration Alkane

v = variable, m = medium, s = strong

Table 3 demonstrates the functional groups of the chloroform: ethanol leaf extract of E. acoroides. The FTIR

spectra are shown in Figure 2. The peaks at 1743 cm−1, 1219 cm−1 and 771 cm−1 were strong, while the others varied

Malaysian Journal of Analytical Sciences, Vol 24 No 1 (2020): 70 - 77

74

from weak to medium. Various compounds were found, including hydroxyl groups, lipids, alkanes, secondary a

mines, benzenoid compounds and phenols.

Figure 2. FTIR spectrum of E. acoroides leaf extract

Table 3. FTIR spectral peak values and functional groups of E. acoroides leaf extract

Spectrum

No.

Wave Number

cm−1

Wave Number

cm−1

[28, 29]

Functional Group Assignment Predicted Compound

1 3612.67 3500–3650 v; O-H stretching vibration (free) Hydroxyl group

2 2931.80 ~2930 m; CH2 asymmetry stretching Lipid

3 2858.51 2800–2900 w; C-H stretching vibration Alkane

4 2401.38 2400–2600 w-m; N-D stretching vibration Imine

5 1743.65 1734–1745 v; C=O of esters Membrane lipid, fatty acid

6 1544.98 1490–1580 w; N-H deformation vibrations Secondary amine

7 1463.97 1440–1465 m; C-H deformation vibration R-CH3; Alkane

8 1219.01 1165–1225 C-C skeletal vibration -C(CH3)3 Alkane

9 771.53 700–900 s; C-H out-of-plane bending (oop)

vibration for substituted benzenes ring

Benzenoid

10 677.01 610–690 w; C-H wagging vibration Vinyl hydrocarbon

compounds

11 422.41 375–450 w; aromatic C-OH in-plane bending vibration

Phenol

v = variable, m = medium, s = strong

As can be seen in Tables 2 and 3, there were differences in the functional groups present between the dried leaf

powder and the chloroform: ethanol (9:1) leaf extract of E. acoroides. The FTIR identified that the leaf extract

contained strong C-H out-of-plane bending (oop bend) vibration for substituted benzene ring, indicating the

Pharmawati & Wrasiati: PHYTOCHEMICAL SCREENING AND FTIR SPECTROSCOPY ON CRUDE

EXTRACT FROM Enhalus acoroides LEAVES

75

presence of phenols and flavonoids in the crude E. acoroides extract. Flavonoids are polyphenols characterised by

two benzene rings joined by a linear carbon chain [30]. The identification of benzenoid compounds via FTIR

spectrophotometry supported the findings from the phytochemical screening, which detected the presence of

phenols and flavonoids. The amines, imines, alkanes and phenols present were considered the major functional

groups of bioactive compounds [31].

The peak around 1734–1745 cm−1 was assigned as C=O ester [31], which may be related to pheophytin and

chlorophyll [32]. In this study, the position of the peak was at 1743 cm−1 (Table 3). The carotenoids were predicted

to be present in the dried leaf powder of E. acoroides. The FTIR spectrum at 1654 cm−1 (Table 2) was assigned as

the C=O conjugate. The conjugated double bond in carotenoids has been reported as the structure responsible for

light absorption [33]. A previous study stated that a peak at 1654 cm−1 is due to chlorophyll and protein content

[34].

The separation of the E. acoroides extract via TLC is presented in Table 4. Eight spots were detected, and the Rf

values ranged from 0.11 to 0.94. The colours were separated into bluish-green, dark green, yellow and yellowish-

grey, and the predicted pigments included chlorophyll b, ethyl-chlorophyllide a, Mg-free chlorophyll b, lutein, Mg-

free chlorophyll a, pheophytin and β-carotene.

Table 4. Identification of compounds based on Rf values and colour from TLC profiling

Fraction

No.

Rf

Value

Colour Colour from

Reference [26]

Identification Maximum

Spectra (nm)

1 0.11 Bluish-green Blue-green Chlorophyll a 655

2 0.21 Dark green Green Chlorophyll b 435 and 645

3 0.24 Bluish-green Blue-green Ethyl-chlorophyllide a 434 and 654

4 0.28 Yellow Yellow Mg-free chlorophyll b 654

5 0.33 Light yellow Yellow Lutein 649

6 0.39 Yellowish-grey Grey Mg-free chlorophyll a 630 and 656

7 0.53 Grey Grey Pheophytin 424 and 658

8 0.94 Orange Yellow-orange β-carotene 425

The maximum spectra for each spot of TLC for fractions one to eight are presented in Table 4. Based on a

comparison to Lichtenthaler and Buschmann [35], fraction five with a maximum absorption at 654 nm was

identified as chlorophyll b, and fraction eight with a maximum absorption at 425 nm was identified as β-carotene.

The other fractions were unidentified. The presence of β-carotene was in agreement with the results from the FTIR

analysis, which confirmed the presence of chlorophyll, pheophytin and carotenoids in the E. acoroides extract,

wherein a functional group of C=O ester was identified (Table 3).

Based on the phytochemical compounds discovered in the FTIR and TLC analyses, E. acoroides has the potential to be used for biomedicinal applications, as phenolic compounds, tannins, flavonoids, chlorophyll and carotenoids are

known to have antioxidant activities [36]. These findings provide an opportunity for the development of natural

products from E. acoroides in drug discovery.

Conclusion

The phytochemical compounds present in the chloroform: ethanol (9:1) leaf extract of E. acoroides were phenols,

tannins and flavonoids. The major functional groups identified were hydroxyl groups, lipids, alkanes, secondary

amines, fatty acids, benzenoid compounds and phenols. The FTIR analysis also identified the presence of

chlorophyll and carotenoids in the extract, which was then confirmed by TLC profiling. Further studies will focus

on the fractionation and purification of potential bioactive compounds present in the extract.

Malaysian Journal of Analytical Sciences, Vol 24 No 1 (2020): 70 - 77

76

Acknowledgement

This research was funded by Ministry of Research, Technology and Higher Education of the Republic of Indonesia

with Grant Number 492.27/UN14.4.A/LT/2019

References

1. Winston, J. C. (1999). Health-promoting properties of common herbs. The American Journal of Clinical Nutrition, 70(3): 491s-499s.

2. Krishnaiah, D., Sarbatly, R. and Nithyanandam, R. (2011). A review of the antioxidant potential of medicinal

plant species. Food and Bioproducts Processing, 89(3): 217-233.

3. Saeed, N., Khan, M. R. and Shabbir, M. (2012). Antioxidant activity, total phenolic and total flavonoid

contents of whole plant extracts Torilis leptophylla L. BMC Complementary and Alternative Medicine, 12: 221.

4. Al-Gubory, K. H. (2014). Environmental pollutants and lifestyle factors induce oxidative stress and poor

prenatal development. Reproductive BioMedicine Online, 29: 17-31.

5. Ighodaro, O. M. and Akinloye, O. A. (2018). First line defence antioxidants-superoxide dismutase (SOD),

catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence

grid. Alexandria Journal of Medicine, 54(4): 287-293.

6. Björk, M., Short, F., Mcleod, E. and Beer, S. (2008). Managing seagrasses for resilience to climate change.

IUCN, Gland, Switzerland: pp. 8-18. 7. Kuriandewa, T. E., Kiswara, W., Hutomo, M. and Soemodihardjo, S. (2003). The seagrasses of Indonesia. In:

Green, E. P. and Short, F. T., Eds., World Atlas of Seagrasses, University of California Press, Barkeley: pp.

172-182.

8. Kuo, J. (2007). New monoecious seagrass of Halophila sulawesii (Hydrocharitaceae) from Indonesia. Aquatic

Botany, 87(2): 171-175.

9. Pharmawati, M., Nurkamila, U. S. and Stevanus. (2016). RAPD fingerprinting key and phylogenetic of nine

seagrass species from Sanur coastal water, Bali, Indonesia using matK sequences. Biodiversitas, 17(2): 687-

693.

10. Ogawa, H. and Namba, N. (2002). Ecological characteristics of tropical seagrasses, especially Enhalus

acoroides. Fisheries Science, 68(sup2): 1767-1770.

11. Amudha, P., Vanitha, V., Mohanasundaram, S, Bharathi N. P. and Jayalakshmi, M. (2017). Phytochemical analysis and invitro antioxidant screening of seagrass Enhalus acoroides. International Journal of Research

and Pharmaceutical Science, 8(2): 251-258.

12. Santoso, J., Anwariyah, S., Rumiantin, R. O., Putri, A. P., Ukhty, N. and Yoshie-Stark, Y. (2012). Phenol

content, antioxidant activity and fiber profile of four tropical seagrasses from Indonesia. Journal of Coastal

Development, 15(2): 189-196.

13. Windyaswari, A. S., Purba, J. P., Nurrahmah, S. S., Ayu, I. P., Imran, Z., Amin, A. A., Kurniawan, F., Pratiwi,

N. T. M. and Iswantari, A. (2019). Phytochemical profile of seagrass extract (Enhalus acoroides): A new

marine source from Ekas Bay, East Lombok. IOP Conference Series: Earth and Environmental Science, 278,

012081.

14. Tuapattinaya, P. M. J. and Rumahlatu, D. (2019). Analysis of flavonoid levels of Enhalus acoroides in different

coastal waters in Ambon Island, Indonesia. International Journal of Applied Biology, 3(1): 70-80.

15. Khan, S. A., Khan, S., B., Khan, L. U., Farooq, A., Akhtar, K. and M. Asiri A. M. (2018). Fourier transform infrared spectroscopy: Fundamentals and application in functional groups and nanomaterials characterization.

In: S. K. Sharma, S. K., Ed., Handbook of Materials Characterization. Springer International Publishing AG,

Springer Nature, Switzerland: pp.317-344.

16. Hemmalakshmi, S., Priyanga, S. and Devaki, K. (2017). Fourier transform infra-red spectroscopy analysis of

Erythrina variegata L. Journal of Pharmaceutical Sciences and Research, 9(11): 2062-2067.

17. Rajiv, P., Deepa, A., Vanathi, D. P. and Vidhya, D. (2017). Screening for phytochemicals and FTIR analysis of

Myristica dactyloids fruit extract. International Journal of Pharmacy and Pharmaceutical Sciences, 9(1): 315-

318.

18. Maobe, M. A. G. and Nyarango, R. M. (2013). Fourier transformer infra-red spectrophotometer analysis of

Urtica dioica medicinal herb used for the treatment of diabetes, malaria and pneumonia in Kisii region,

Southwest Kenya. World Applied Sciences Journal, 21(8): 1128-1135.

Pharmawati & Wrasiati: PHYTOCHEMICAL SCREENING AND FTIR SPECTROSCOPY ON CRUDE

EXTRACT FROM Enhalus acoroides LEAVES

77

19. Kumar, S., Jyotirmayee, K. and Sarangi, M. (2013). Thin layer chromatography: A tool of biotechnology for

isolation of bioactive compounds from medicinal plants. International Journal of Pharmaceutical Science

Review and Research, 18(1): 126-132.

20. Porika, R., Poojari, S., Lunavath, V. and Mamidala, E. (2014). Preliminary phytochemical investigation and

TLC analysis of P. angulata fruit extract. IOSR Journal of Pharmacy and Biological Sciences, 9(2): 11-14.

21. Forgacs, E. and Cserhati, T. (2002). Thin-layer chromatography of natural pigments: New advances. Journal of Liquid Chromatography and Related Technologies, 25(10-11): 1521-1541.

22. Zahra, N., Alim-un-Nisa, Fatima, Z., Kalim, I. and Saeed, K. (2015). Identification of synthetic food dyes in

beverages by thin layer chromatography. Pakistan Journal of Food Science, 25(4): 178-181.

23. den Hartog, C. and Kuo, J. (2006). Taxonomy and biography of seagrasses. In Larkum, T., Orth, R. J. and

Duarte, C. M., Eds. Seagrasses: Biology, ecology and conservation. Springer, The Netherlands: pp. 1-23.

24. McKenzie, L J. and Yoshida, R. L. (2009). Seagrass-watch. Proceeding of a workshop for monitoring seagrass

habitats in Indonesia. The Nature Conservancy, Coral Triangle Center, Sanur, Bali: pp. 29-32.

25. Harborne, J. B. (1984). Phytochemical methods: A guide to modern technique of plant analysis. (2nd ed).

Chapman and Hall. London. pp. 37-168.

26. Bacon, M. F. and Holden, M. (1967). Changes in chlorophylls resulting from various chemical and physical

treatments of leaves and leaf extracts. Phytochemistry, 6: 193-210.

27. Dewi, C. S. U., Kasitowati, R. D. and Siagian, J. A. (2018). Phytochemical compounds of Enhalus acoroides from Wanci Island (Wakatobi) and Talango Island (Madura) Indonesia. IOP Conference Series: Earth and

Environmental Science, 137: 012045.

28. Spectroscopic Tools. Access from http://www.science-and-fun.de/tools/. [Access online 21-25 October 2019]

29. Socrates, G. (2004). Infrared and Raman characteristic group frequencies tables and charts. John Wiley &

Sons Ltd, Baffins Lane, Chichester, West Sussex PO19 IUD, England: pp. 1-340.

30. Corcoran, M. P., McKay, D. L. and Blumberg, J. B. (2012). Flavonoid basics: Chemistry, sources, mechanisms

of action, and safety. Journal of Nutrition in Gerontology and Geriatrics, 31(3): 176-189.

31. Pratheeba, T., Ragavendran, C. and Natarajan, D. (2015). Larvicidal, pupicidal and adulticidal potential of

Ocimum gratissimum plant leaf extracts against filariasis inducing vector. International Journal of Mosquito

Research, 2 (2): 1-8.

32. Li, X., Zhou, R., Xu, K., Xu, J., Jin, J., Fang, H. and He, Y. (2018). Rapid determination of chlorophyll and pheophytin in green tea using Fourier transform infrared spectroscopy. Molecules, 23: 1010.

33. Mezzomo, N. and Ferreira, S. R. S. (2016). Carotenoids functionality, sources, and processing by supercritical

technology: A review. Journal of Chemistry, 2016: 1-16.

34. Kushwaha, K., Saxena, J., Tripathi, B. K. and Agarwal, M. K. (2014). Detection of carotenoids in

psychrotrophic bacteria by spectroscopic approach. Journal of BioScience and Biotechnology, 3(3): 253-260.

35. Lichtenthaler, H. K. and Buschmann, C. (2001). Chlorophylls and carotenoids: Measurement and

characterization by UV-VIS spectroscopy. In: Wrolstad, R. E., Acree, T. E., An, H., Decker, E. A., Penner, M.

H., Reid, D. S., Schwartz, S. J., Shoemaker, C. F. and Sporns, P., Eds., Current protocols in food analytical

chemistry (CPFA), John Wiley and Sons, New York: pp. F4.3.1-F4.3.8.

36. Lachowicz, S., Oszmiański, J. and Wiśniewski, R. (2018). Determination of triterpenoids, carotenoids,

chlorophylls, and antioxidant capacity in Allium ursinum L. at different times of harvesting and anatomical

parts. European Food Research Technology, 244(7): 1269-1280.