The Levels of Quercetin and Antioxidant Activity of Patikan Kebo

Leaves Extract

Sherly Asri Widyaningrum

, Brian Wasita

and Adi Magna Patriadi Nuhriawangsa

Postgraduate Program of Nutrition Sciences, School of Postgraduate, Sebelas Maret University, Surakarta, Indonesia

Department of Anatomical Pathology, Faculty of Medicine, Sebelas Maret University, Surakarta, Indonesia

Department of Animal Science, Faculty of Agriculture, Sebelas Maret University, Surakarta, Indonesia

Keywords: Quercetin, Antioxidant Activity, Patikan Kebo Leaves Extract.

Abstract: Patikan kebo is a wild plant that is commonly found in tropical and subtropical areas on soil that is not so

moist. Patikan kebo contains a number of active compounds such as flavonoids, terpenoids, and several other

active compounds such as alkaloids and polyphenols. This study aims to determine the concentration of

quercetin and antioxidant activity in patikan kebo leaf extract. Samples were taken from the Swadaya Alam

Jaya Herbal Research Center for Medicinal Plants, and bioactive compounds were extracted from the leaves

of the patikan kebo plant through the mesarasi method with ethanol. Quercetin levels were analyzed using the

High-Performance Liquid Chromatography (HPLC) method, and antioxidant activity was carried out using

the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. The results showed that the patikan kebo leaf extract

showed that the content of quercetin compounds in 100 g of the patikan kebo leaf extract was 110.73 mg and

the IC

antioxidant activity results were 80,151 % and 79,962 %, respectively.

1 INTRODUCTION

Indonesia is a tropical island and is famous for its

natural wealth with various types of nutritious plants

that have properties (Widjaja et al., 2014). Globally,

there are 40,000 types of plants are known as

traditional medicines, 30,000 of which are in

Indonesia, of which 7000 types of plants have been

used as raw materials for herbal medicines. One of

them is the patikan kebo plant (Jumiarti and

Komalasari, 2017).

Patikan kebo is a wild plant that is easily found on

the surface of the soil that is not so moist, and contains

a number of active compounds such as flavonoids,

terpenoids, and a number of other active compounds

such as alkaloids and polyphenols (Asha et al., 2014).

The important bioactive components of patikan kebo

are flavonoids, which function as antioxidants.

Several studies have shown that antioxidant

compounds can play a role in fighting oxidative

damage through their reactions with free radicals in a

way that correlates with oxidative stress defense so

that antioxidant compounds can play a role in fighting

oxidative damage (Uppala and Reddy, 2014). One of

the flavonoid groups found in the patikan kebo plant

is quercetin (Huang et al., 2012) (Karim, Jura, &

Sabang, 2015). Quercetin may protect against the

environmental causes of free radicals (David et al.,

2016). The benefits of antioxidants and quercetin for

health are the basis for exploring plants or foods that

contain these compounds. One of them is the patikan

kebo plant.

The bioactive components in patikan kebo leaves

can be increased by extraction because this method

can attract and separate several compounds using

certain solvents (Leba, 2017). The extraction methods

that are widely used for solid samples into solvents

are soxtellation, percolation, and maceration. The

maceration method is a simple extraction method that

is most commonly used for the extraction of patikan

kebo. This method can also avoid the risk of damage

to compounds in plants (Tetti M, 2014).

Quercetin compounds and antioxidant activity

can be dissolved in polar solvents such as ethanol,

methanol, ethyl acetate, butanol, potreleum, and

others (Tran, 2020). So far, there have been no studies

analyzing compounds in patikan kebo leaves taken

from local soil in Gianyar, Bali, which have the

potential to contain quercetin and antioxidant activity.

This research is about the content of these compounds,

which can be used as empirical evidence that patikan

kebo leaf extract can be used for health. Based on the

Widyaningrum, S., Wasita, B. and Nuhriawangsa, A.

The Levels of Quercetin and Antioxidant Activity of Patikan Kebo Leaves Extract.

DOI: 10.5220/0011650500003608

In Proceedings of the 4th International Conference on Social Determinants of Health (ICSDH 2022), pages 89-93

ISBN: 978-989-758-621-7; ISSN: 2975-8297

 2023 by SCITEPRESS – Science and Technology Publications, Lda. Under CC license (CC BY-NC-ND 4.0)

above background, this study aims to determine the

concentration of quercetin and antioxidant activity in

patikan kebo leaf extract.

2 MATERIALS AND METHODS

2.1 Methods

This study used patikan kebo leaves, which were

taken homogeneously in the local area by the Jaya

Herbal Self-Help Research Center for Medicinal

Plants, Gianyar Regency, Bali, Indonesia. The

extraction of patikan kebo leaves was carried out at

the Center for Food and Nutrition Studies, Gadjah

Mada University (UGM), and the evaluation of

quercetin levels and antioxidant activity was carried

out at the Chemical Laboratory of the University of

Muhammadiyah Malang (UMM).

Instruments and Materials

The tools used in this study were digital scales, test

tube glass, UV-Vis spectrophotometer, Whatman no.

1 filter paper, shaker and rotating evaporator, mortar

hammer, measuring pipette, suction cup, measuring

flask, glass funnel, erlenmeyer, beaker, test tube,

cuvette. The main ingredients used in the

manufacture of patikan kebo leaves extract are

patikan kebo leaf simplisa and 96% ethanol.

Methanol p.a, DPPH 0.2 mM (Dissolve as much as

0.078 g in methanol p.a to 1000ml volume), distilled

water, H3Po4 solution, and 25% acetonitrile.

Extraction of the Patikan Kebo

The patikan kebo leaves that have become simplisa

are soaked in a 96% ethanol solution with a ratio of 1

g simplisa to 10 ml ethanol for ± 48 hours while

stirring regularly using a shaker at 150 rpm at 600 C.

After going through the soaking process, the patikan

kebo leaves are filtered The filtrate was taken using

Whatman filter paper number 1. Then the filtrate was

inserted into a rotatory evaporator to make it more

concentrated and obtain patikan kebo leaves extract.

Evaluation of the Total Quercetin

Analysis of quercetin levels was carried out using the

HPLC method on a Shimadzu UFLC system,

equipped with a LC 20AT quaternary gradient pump

using an SPD-M20A PDA diodearray detector

(DAD). The data were obtained on a liquid

chromatography solution administrator data system

Evaluation of the Antioxidant Activity

For the determination of antioxidant activity, first

sample preparation, i.e. sample taken 0.1 ml dissolved

in methanol pa to 100 ml in a measuring flask, to

obtain a sample solution of 100 ppm, then

homogenized and allowed to stand for about 30

minutes, then filtered and if necessary centrifuged at

3000 rpm for 10 minutes then supernatant was taken.

Next, 1.5 ml of the sample solution was added with 3

ml of 0.2 mM DPPH solution, then the mixture was

shaken until homogeneous and allowed to stand for

30 minutes, then the absorbance was measured at 516

nm. The procedure was carried out on a blank of 1.5

ml of methanol pa. The antioxidant activity of the

sample could be determined by calculating the

percentage of DPPH absorption inhibition with the

formula:

Antioxidant Activity (%) =



1 −







𝑥 100%

Information :

b = Absorbance Blank

s = Absorbance Test Sample

3 RESULTS AND DISCUSSION

This study aims to determine the concentration of

quercetin and antioxidant activity in patikan kebo leaf

extract. Samples were taken from the Jaya Herbal

Bali Self-Help Research Center for Medicinal Plants

and extracted using the maceration method with

ethanol as a solvent to obtain the samples used in this

study. The advantage of using the maceration method

is that there is no heating process, so it is unlikely that

the active substances in simplisa are damaged or

decomposed (Istiqomah, 2013). The solvent used in

this study was ethanol. Extraction solvents such as

ethanol, methanol, dichloromethane, ethyl acetate,

water, and hexane are widely used to extract

antioxidants and other compounds, but ethanol

solvents have a high polarity when compared to other

types of organic solvents. Ethanol has a low boiling

point. Ethanol is also non-toxic and dangerous, so it

tends to be safe (Bakhouche et al., 2015; Hoon et al.,

2015).

The patikan kebo leaf extract contains a number

of phenolic and flavonoid compounds (Asha et al.,

2014), one of which is quercetin. The Patikan kebo

leaves extract contains a number of phenolic and

flavonoid compounds (Asha et al., 2014), one of

which is quercetin. Quercetin is a compound

classified as a flavanol, which is the main polyphenol

flavonoid found in various plants (Anand et al., 2016).

These compounds have anticancer, antiviral, and anti-

ICSDH 2022 - The International Conference on Social Determinants of Health

inflammatory properties, as well as the ability to treat

metabolic disorders, cardiovascular disease, arthritis,

allergies, and inflammation (Batihah et al., 2020).

Quercetin can also control postprandial blood sugar

levels by activating α-glucosidase enzyme inhibitors

(Jadhav, 2012). In this study, the levels of quercetin

were identified using the HPLC method, which is a

method of separating a mixture of substances using a

mobile and stationary phase, the separation technique

in HPLC occurs because of differences in absorption

power, solubility, partitioning, molecular size, ion

size, and vapor pressure on the components carried by

the mobile phase through the stationary phase (Aulia

et al., 2016).

Table 1: Results of analysis of quercetin levels using HPLC.

Sampel ID RT min Result (µg/g)

atikan kebo leaf extrac

6,739 110726,51549

Based on the results, it can be seen that the quercetin

compound was seen at a retention time of 6,739

minutes. From the results of this study, it can be seen

that the patikan kebo leaf extract contains quercetin at

a concentration of 110.73 mg/gr. The ethanol solvent

used is able to attract quercetin compounds, because

kuers is a flavonoid group compound and can be

dissolved in ethanol solvents (Sukmawati et al., 2019).

Factors that affect the production of secondary

metabolites are divided into two factors, namely

extrinsic and intrinsic. Extrinsic in the form of

climatic conditions and the soil where it grows, while

the intrinsic factors are in the form of the genes of the

plant concerned, as well as climatic conditions, which

include temperature, rainfall, lighting, and soil

elevation, while soil conditions include the content of

nutrients, minerals, moisture, pH, rocks, sand, and

mud (Liu et al., 2017).

Previous studies reported that having tested

quercetin levels in patikan kebo leaf extract with

various solvents showed results of 276.6 mg/gr, 189.2

mg/gr, 164.1 mg/gr, 124.3 mg/gr, and 84,8 mg/gr

(Tran, 2020). The difference in quercetin value results

depends on differences in plant species, solvents and

concentrations, as well as the method used at the time

of extraction (Hardinsyah et al., 2019).

The antioxidant activity test of patikan kebo

leaves extract was identified using the DPPH method,

because the test is simple, fast, and easy and only

requires a small sample (Shalby et al., 2013). The

working principle of DPPH (2,2-Diphenyl-1-

picrylhydrazyl) is that antioxidant compounds will

donate hydrogen atoms to DPPH when the DPPH

solution reacts with antioxidant compounds. Then,

measured by UVVis at a wavelength of 516nm, there

was a color change (from dark purple to yellow or

pale yellow) when the extract sample showed the

ability to absorb DPPH free radical activity (Kedare

et al., 2011). The results of the research on

antioxidant activity in patikan kebo leaves extract can

be seen in the following table.

Table 2: Antioxidant Activity of Patikan Kebo Leaf Extract

(%) concentration 100 ppm.

ppm concentration ul Abs

%antioxida

nt activit

atikan kebo leaf extrac

1 0.105 80,151

2 0.106 79,962

Based on table 2, patikan kebo leaf extract has

antioxidant activity of 80.151% and 79.962%,

respectively. The ability of antioxidants as free

radical scavengers is associated with the ability of

these antioxidants as proton donors. The number of

hydrogen protons that can be donated is influenced by

the number and position of the aromatic hydroxyl or

hydroxyl groups of the phenolic component. The

more aromatic hydroxyl groups, the more effective

the chain reaction inhibition in the oxidation process

is by donating hydrogen atoms or acting as free

radical acceptors (Septiana & Asnani, 2013).

Research conducted by Karim, K et al., (2015)

reported the optimum results of antioxidant activity

in patikan kebo leaf extract of 99.21%. Antioxidants

are said to be very strong if they have an IC

value

of less than 50 ppm, strong categories range from 50-

100 ppm, moderate antioxidants are 100-150 ppm,

and weak antioxidants are in the range of 150-200

ppm. al., 2017). The smaller the IC

value of a

compound, the more effective the compound is as an

antioxidant (Putri et al., 2013). The difference in

antioxidant activity values obtained from each type of

extract may be due to differences in the content and

number of active compounds contained in the extract

(Purwanto et al., 2017).

The level of polarity of the solvent also greatly

affects the antioxidant activity. In addition, the

decrease in temperature during storage can also affect

the levels of phytochemicals or antioxidant activity

(Li et al., 2012). Plants that have high antioxidant

bioactive compounds can help the body prevent

damage by radical compounds or decrease body

resistance (Mensaah et al., 2014).

The results of the analysis of phytochemical

compounds in patikan kebo leaf extract proved to

contain quercetin and strong antioxidant activity.

Therefore, to continue the potential pharmacological

use of patikan kebo leaf extract, it is necessary to

conduct appropriate preclinical research.

The Levels of Quercetin and Antioxidant Activity of Patikan Kebo Leaves Extract

4 CONCLUSIONS

Based on the results of the research, it was concluded

that the patikan kebo leaf extract contained a

quercetin compound of 110.73 mg/gr. The

antioxidant activity obtained from patikan kebo

leaves extract has a strong antioxidant activity

category.

ACKNOWLEDGEMENTS

In particular, the authors would like to thank the

Center for Food and Nutrition Studies Laboratory,

Gadjah Mada University (UGM) and Chemistry

Laboratory, University of Muhammadiyah Malang

(UMM) who have helped the authors in collecting

data, as well as my supervisor who has taken the time

to provide guidance.

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The Levels of Quercetin and Antioxidant Activity of Patikan Kebo Leaves Extract