Evaluation of Antioxidant and Cytotoxicity Activity of Cyperus

rotundus L. Rhizome Extract and Fractions

Masfria

, Urip Harahap

, Denny Satria

Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Sumatera Utara,Medan, Indonesia

Department of Pharmacology Faculty of Pharmacy, Universitas Sumatera Utara,Medan, Indonesia

Doctoral Programme, Faculty of Pharmacy, Universitas Sumatera Utara,Medan, Indonesia

Keywords: antioxidant, cytotoxicity, Cyperus rotundus L., rhizome

Abstract: Excess oxygen as free radicals and unbalanced antioxidant protection capabilities result in many

diseases such as breast cancer. The purpose of this study was to evaluate the radical activity, total

phenolic and cytotoxicity on 4T1 cell lines extracted from ethanol (EE), n-hexane (nHF), ethylacetate

(EAF) and residual (RF) fractions of Cyperus rotundus L. rhizomes. maceration method to obtain

ethanol extract and fractionated with ethylacetate and n-hexane. DPPH method to measure antioxidant

activity, colorimetric method to measure total phenolic content, and MTT assay to measure cytotoxicity

activity. Antioxidant activity from DPPH assay measured as IC

were 41.12 ± 0.03; 212.43 ± 0.10;

57.34 ± 0.03; 194.08 ± 0.05 µg/mL respectively.Total phenolic content measure were 138.25 ± 0.73;

51.93 ± 1.71; 213.01 ± 2.83; 111.07 ± 0.68 mg/GAE respectively. Cytotoxic activity were found to have

of139.92 ± 0.91; 125.71 ± 0.57; 3685.36 ± 201.93; 3999.98 ± 437.39µg/mL respectively. The

results reveal that Cyperus rotundus L.rhizomes haveantioxidant and cytotoxicity activities. Our further

study is to assess the anticancer mechanism of Cyperus rotundus L. rhizomes.

1 INTRODUCTION

Reactive oxygen species (ROS) such as superoxide

anions, singlet oxygen, hydrogen peroxide and

hydroxyl are always produced in biological reactions

or from the surrounding environment (Nagulendra,

2007). Spesies reaktif adalah molekul yang memiliki

fluiditas elektronik dan sangat reaktif. Produksi

radikal bebas oksigen yang berlebihan dan

mekanisme pertahanan antioksidan yang tidak

seimbang menyebabkan banyak penyakit, terutama

penuaan dan kanker (Yang, 2004) (Nagmoti, 2012)

(Jamuna, 2012) (Rosidah, 2008).

Breast cancer is the most common in the 1st

estimated incidence of cancers, and 2nd highest

incidence of death occurs in women who have

cancer in the USA published in recent research

(Siegel, 2015).

Cyperus rotundus L. has been traditionally

used as a medicine for abdominal pain, menstrual

irregularities, and intestinal disorders, (Peerzada,

2015). Its extract is used as antimutagenic, radical

scavenging, insecticide and volatile oils as

antibacterial and cytotoxic based on the results of

bioassay (Hadi, 2007) (Memariani, 2017)

(Vitaglione, 2004) (Lanciotti, 2004) (Kilani, 2008)

(Tenore, 2011) (Nam, 2016) (Heman, 2014) (Sonwa

and Konig, 2001) (Liu, 2016) (Kilani, 2007)

(Jirovetz, 2004) (Liu, 2016). The aim of our research

was to assess the antiproliferative and antioxidant

effect of ethanol extract from Cyperus rotundus L.

rhizome.

2 MATERIALS AND METHODS

2.1 Plant and Chemicals Material

Extract and fractions were obtained from Masfria.

The chemicals used are sodium bicarbonate (Merck),

Dymethylsulfoxide (Sigma), DPPH (TCI), MTT salt

(Sigma), folin ciocalteu (Sigma), and distilled water.

Masfria, ., Harahap, U. and Satria, D.

Evaluation of Antioxidant and Cytotoxicity Activity of Cyperus Rotundus L. Rhizome Extract and Fractions.

DOI: 10.5220/0010092208210824

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

821-824

ISBN: 978-989-758-449-7

821

2.2 Preparation of Extract

Extraction with ethanol 96% and fractionation were

followed by previous study (Satria, et al., 2015:

Anggraeni, et al., 2014; Hasibuan, et al., 2015).

2.2.1 Free Radical Scavenging Activity

Assay

Free radical scavenging activity was determined

based on (Satria, 2017) procedure and IC

value

was described with regression analysis

2.2.2 Determination of Total Phenol

Concentration

Briefly, 0.1 mL extract and fraction (500 μg / ml)

were used in analysis and the procedure based on

(Rosidah, et al., 2008). The concentration is

expressed as GAE in mg per gram of extract.

(Jamuna, 2012) (Sitorus, 2017).

2.3 Cytotoxicity Assay

The cells were exposed with ethanol extract, n-

hexane, ethylacetate, residual fraction. In this test,

4T1 cell lines were seeded in DMEM medium, and

the procedure was followed by previous study

(Harahap, 2018). The data absorbed from each well

is converted into viable percentage of cells (Satria,

2014) (Dalimunthe, 2018).

2.4 Statistical Analysis

Load data as mean ± SD. SPSS 22 software is used

to analyze all statistics.

3 RESULTS AND DISCUSSION

3.1 Radical Scavenging Activity

DPPH, which is a stable free radical, is centered on

nitrogen and produces a deep purple color in

methanol solution so that it can be used to test the

antiradical effect of plant samples determined in

terms of the ability of the active compound to

contribute to hydrogen. Both antioxidants transfer

electrons or hydrogen atoms to DPPH, thus

neutralizing the freeradical character. The DPPH test

has been widely used as a fast, accurate and

reproducible parameter for finding high antioxidant

activity from pure compounds from plant extracts

(Koleva, 2002) (Meir, 1995). Low IC

values reflect

high fraction antioxidant activity, because

concentrations are important for inhibiting radical

oxidation at 50% low. IC50 for ethanol extract, n-

hexane, ethylacetate, the residual fraction in DPPH

test was 41.12 ± 0.03; 212.43 ± 0.10; 57.34 ± 0.03;

194.08 ± 0.05 µg / mL respectively.

3.2 Total Phenolicand Total Flavonoid

Contents

The Folin Ciocalteau method for measuring total

phenolic content (TPC) claimed by based on the

reduction of the phosphomolybdic-phosphotungstate

to the blue colour in alkaline solution (Cicco, 2009).

Ethanol, n-hexane, ethylacetate, residual fraction of

Cyperus rotundus L. were found to contain various

levels of phenolic content. 138.25 ± 0.73; 51.93 ±

1.71; 213.01 ± 2.83; 111.07 ± 0.68 mg GAE /

grespectively.

Phenolic compounds are recognised as

antioxidant (Shahidi and Wana Sundara, 1992) and

they are compounds in plants which important

cause of their radical scavenging ability due to their

hydroxyl groups. (Hatano, 1989) (Diab, Reham and

Shin, 2015) (Dai and Mumper, 2010) (Heim,

Tagliaferro and Bobilya., 2002).

3.3 Inhibitory Concentration 50%

(IC

)

The percentage of cell viability was measured by the

MTT method after being stored for 24 hours. In each

n-hexane treatment, ethanol extract, ethylacetate,

residual fraction represents inhibition of cell growth.

IC50 value of ethanol extract, n-hexane, ethyl

acetate, residual fraction was 139.92 ± 0.91; 125.71

± 0.57; 3685.36 ± 201.93; 3999.98 ± 437.39 μg /

mL, respectively. Cytotoxicity estimates of natural

products related to the level of active compounds in

this plant include Cyperus rotundus L.

These plants contain coumarin flavonoids,

monoterpene, sesquiterpenes, saponins, and steroids

which can be measured as active substances

(Nidulaga, 2017) (Nidigula, 2016) (Yadav, 2010).

ICOSTEERR 2018 - International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches

822

ACKNOWLEDGEMENTS

This research was funding through “Hibah

Penelitian Guru Besar 2018” from University of

Sumatera Utara.

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