Article

Antioxidant Activity of Bioactive Compounds in the Ethyl Acetate Extract of

Putat Leaves (Planchonia valida)

Nelson^1*, Desy Vania Silaban¹, Faizar Farid¹, Ratih Dyah Puspitasari¹, Indra Lasmana Tarigan^1,2

Ilham Ifandi Ramadhan¹, Madyawati Latief^1,2

,

¹Department of Chemistry, Faculty of Science and Technology, Universitas Jambi, Indonesia

²Natural Product and Bioactive Compound Laboratory, Faculty of Science and Technology, Universitas Jambi,

Indonesia

Abstract

The Putat leaf (Planchonia valida) is a plant traditionally used by the community in the Tanjung Lanjut area, Jambi,

both as a salad ingredient and as a traditional medicine to treat skin diseases and promote health. This identifies

that the putat plant contains chemical compounds that potentially have antioxidant activity. In this study, putat leaf

samples (Planchonia valida) that potentially have antioxidant activity. In this study, Planchonia valida (P. valida) leaf

samples using n-hexane and ethyl acetate solvents through a multistep maceration process. Subsequently,

phytochemical screening and antioxidant activity tests were conducted to obtain information about fractions that

have the potential to be further isolated. The extraction percentage with a yield value of 1,28% was obtained in the

maceration process. Further isolation was carried out using isolate liquid chromatography and FT-IR instruments.

The obtained isolate was in the form of a wet solid, and through characterization, it was classified as a steroid

compound suspected to be ergosterol. Free radicals or oxidants have reactive effects on the human body and can

trigger various types of diseases. Antioxidants are known to have the ability to inhibit or neutralize free radicals in

the human body. This study tested antioxidant activity using the DPPH (1,1-diphenyl-1,1-picrylhydrazyl) method on

isolating F1. Isolate F1 was found to have antioxidant activity with a value of 714.637 ppm, falling into the range of

weak antioxidant activity

Keywords: Antioxidant, Bioactive Compounds, Planchonia valida

Graphical Abstract

*

Corresponding author

Email addresses: nelson@unja.ac.id

DOI: https://doi.org/10.22437/chp.v6i4.41632

(https://creativecommons.org/licenses/by/4.0 )

160

Chempublish Journal, 6(4) 2022, 160-167

Introduction

isolated that have antibacterial activity against

(+)-galokatekin, galokatekin-(4α→8)-galokatekin,

α-dimorphphenolic acid, hiptatatic acid, 3-β-

Otrans-p coumaroyltormentic acid, 3-β-O-cis-p-

kumaroyltormentic acid.

Indonesia is a country that has a high wealth of

natural resources. Various plant species thrive in

this country, one of which is the putat plant.

Planchonia valida or better known as putat can be

seen in the Lake Tangkas area and is the most

dominant plant compared to various types of

plants. Based on Jamiat et al (2019), putat plants

are woody plants with taproots, spherical stems

and have branches. Putat plants are theoretically

generally found in swampy areas that are

inundated by water or sometimes inundated by

water (Syukur, 2016).

Antioxidants are defined as compounds that can

delay, slow down and inhibit the process of lipid

oxidation. This compound can reduce the

negative influence of free radicals. Free radicals

are highly reactive molecules, which can disrupt

cell integrity, can react with cell structure

components, can react with cell structure

components such as enzin and DNA. In the body,

free radicals are continuously formed. This leads

to the formation of new, more reactive free

radicals, causing cell damage and death.

Therefore, antioxidants are needed in order to

protect the body from free radicals and reduce

their negative impacts (Handayani, 2020). In

order to fulfill the issues, the search for natural

antioxidant compounds is directed at natural

resources (Ipandi et al., 2016).

Putat leaves can be used to treat skin diseases

such as itching by pounding the leaves then

attached to the itchy area. Putat leaves are also

used as a mixture of postpartum bath water. In

addition to being a medicine, generally people

use putat leaves as a mixture of cold powder and

used to protect the skin from sun exposure when

in the field and can eliminate dark spots on the

face. (Supriningrum et al., 2019). This identifies

that putat plants contain chemical compounds

that are bioactive.

Free radicals in the body are very dangerous

materials. Free radical material is actually a

compound or molecule containing one or more

unpaired electrons in its outer orbital. The

presence of unpaired electrons results in the

compound being very reactive to find a partner.

You do this by binding or attacking the electrons

of molecules around it. Free radicals are

generally bound to large molecules such as lipids,

proteins, and DNA (carriers of traits). If this

happens, it will result in cell damage or cell

growth that cannot be controlled (Leksono,

2018).

Based on research conducted by Shaumi (2019),

through phytochemical screening of putat leaf

shoot powder (Planchonia valida) stated that it

positively contains all chemical compounds,

namely alkaloids, tannins, flavonoids, steroids,

and saponins. This statement is also supported

by research conducted by Supriningrum et al.,

(2019) which explains that alkaloids, flavonoids,

tannins, saponins and steroids are among the

chemical components contained in putat leaves.

Putat leaves have many useful ingredients for the

body that can be used as antibacterials. Plants

that contain antibacterial are usually used as

medicinal plants. Most medicinal plants are

plants that are often used for traditional

medicine and have even been widely managed as

modern medicine.

According to Syamsudin et al., (2019), research

from the genus Planchonia that has been reported

includes phytochemical tests there are saponin

compounds acylated with triterpenoids from

Chemically, free radicals are compounds that

have unpaired electrons, which often look for

other molecules to be stable. Free radicals or

oxidants can have an effect on cell damage in the

human body due to oxidation reactions (Hasanah

et al., 2017). According to Iglesias et al (2016), the

DPPH test is considered more sensitive for

determining

the

antioxidant

activity

of

compounds that are less polar. According to

Karadag (2009) in Wulansari (2018), the DPPH

method has the advantage that the analysis

method is simpler, faster, easier to observe and

Planchonia

careya

species.

6

chemical

compounds from Planchonia careya have been

160

Chempublish Journal, 6(4) 2022, 160-167

more sensitive to samples with relatively small

concentrations so that the results obtained are

more accurate (Wahdaningsih et al., 2013).

a blender and filtered using a 60 mesh sieve so

that putat leaf simplisia is produced. A dry

sample of 1.4 kg was macerated in stages.

Multilevel maceration is performed to separate

the compounds in the sample according to their

degree of polarity. The initial solvent used for

maceration is n-hexane as much as 1.5 L. Then

maceration is continued with ethyl acetate

solvent. Stirring is carried out for 3 hours then

allowed to stand for 48 hours. The extract

obtained from ethyl acetate solvent is then

concentrated using a rotary evaporator. From

ethyl acetate extract, dry extract was obtained as

much as 16.8 grams with a yield of 1.28%.

The results of research conducted by Pratiwi et al

(2019), obtained data showing that the ethyl

acetate fraction has better antioxidant activity

than n-hexane fraction extract. Antioxidant

activity comes from compounds found in the

fraction of red dragon fruit skin among which are

known to be positive based on phytochemicals

are

alkaloids,

terpenoids,

phenolics

and

flavonoids. Tests were carried out on the n-

hexane fraction, ethyl acetate fraction and

methanol fraction, it was found that in the n-

hexane fraction, ethyl acetate fraction and

methanol fraction had less active antioxidant

activity with an IC₅₀value of n-hexane fraction of

198.05μg/ml while in the ethyl acetate fraction of

Phytochemical

analysis was performed to identify various

bioactive compounds, including alkaloids,

screening.

Phytochemical

flavonoids, tannins, saponins, and steroids. A

positive result was characterized by a distinct

199.527μg/ml

and

methanol

fraction

of

color change and/or the formation of

a

445.255μg/ml.

precipitate, signifying a chemical interaction

between the reagent and the plant extract.

Material and Methods

Materials and Instrumentations

The detection of alkaloids involved the addition

of 2N hydrochloric acid (HCl) to the sample,

followed by treatment with Mayer's reagent,

which resulted in the formation of a yellow

precipitate. Furthermore, Bouchardat reagent

induced a red coloration, whereas Dragendorff

The materials used are Putat Leaves (Planchonia

valida) obtained from Tanjung Lanjut Village,

Muaro Jambi regency, Indonesia. Chemical were

used in this research Ethyl Acetate, Aquadest

(Sigma-Aldrich), N-Hexan, Silica Gel G60 (0.063-

0.200 mm and 0.040-0.063 mm), H_2SO42N, HCl

2N, Mayer Reagent, Dragendrof Reagent,

Bouchardat Reagent, HCl, Magnesium powder,

DPPH powder, Standard solution, Vitamin C

reagent

generated

a

brown

precipitate,

confirming the presence of alkaloids (Ningrum et

al., 2016). The presence of flavonoids was verified

through the Flavonoid Test, where the ethyl

acetate extract of putat leaves exhibited an

orange coloration, indicative of flavonoid

compounds (Vanessa et al., 2014). The Tannin

Test yielded a blackish-green coloration upon the

addition of 1% FeCl3 solution, signifying a

positive result. Similarly, the presence of

saponins was confirmed by the formation of

stable, persistent foam, which remained intact

even after the addition of 2N HCl (Wardana and

Tukiran, 2016). The detection of steroids was

established through the Steroid Test, in which

treatment with Liebermann-Burchard reagents

(anhydrous acetic acid and concentrated sulfuric

(ascorbic

instrumentation

evaporator

Chromatography

acid),

Methanol

p.a..

vacuum

Vacuum

Thin

The

rotary

Liquid

Layer

used

RV

are

10),

(IKA

(KVC),

(KLT),

UV-Vis

Spectrophotometry (Thermo Fisher Scientific,

Singapore), FT-IR Spectrophotometry (Thermo

Fisher Scientific), as well as other equipment such

as analytical balances.

Methods

Sample Preparation and Extraction. The sample

to be studied is the leaf part of the putat plant.

The sample is washed with water and dried by

aeration. Next, the putat leaves are mashed with

acid) produced

characteristic

Additionally, the test differentiated terpenoids,

which exhibited an orange or purple-red

a

of

bluish-green coloration,

steroid compounds.

161

Chempublish Journal, 6(4) 2022, 160-167

coloration, from steroids that displayed a bluish-

green hue (Sangi et al., 2013).

IC₅₀value, indicating the concentration at which

50% inhibition is achieved.

Isolation Active Compound. Liquid vacuum

column chromatography (KVC) was performed

using stationary phase silica gel with sample

ratio: silica gel (1:20). The sample extract is

impregging using silica gel, then added to the

column that has contained the stationary phase.

While the mobile phase used is n-hexane: ethyl

acetate and ethyl acetate: methanol with

variations in ratio (10:0; 9:1; 8:2; 7:3; 6:4; 5:5; 4:6;

3:7; 2:8; 1:9; and 0:10). The fraction obtained is

accommodated and then evaporated. The results

of column chromatography were performed

again KLT. Silhouettes that have identical smudge

patterns are combined based on the Rf value on

the chromatogram. Eluents that have one spot

stain are then tested using 3 different eluents,

where if KLT remains one spot stain then isolates

are obtained.

Data Analysis.

Determination of %Yield. Determination of the

percentage of extract yield was calculated using

equation 1.

Mass of Extract (g)

%Yield =

× 100%

(1

Mass of Simplicia (g)

Determination of Antioxidant Performance.

The determination of the % inhibition value is

done using the equation 2.

% inhibition =

control absorbance−sample absorbance

x 100%

(2)

control absorbance

IC₅₀Value. The IC₅₀value can be obtained by

plotting a graph of the relationship of sample

concentration with % inhibition. The x-axis is the

sample concentration while the y-axis is

%inhibition. Then calculated the regression

equation y = a + bx where to calculate the value

of IC₅₀can be known from replacing the value of

y in the regression equation with the value of 50.

Antioxidant activity. The preparation of the

DPPH solution involves dissolving 1.97 mg of

DPPH powder into 100 mL of methanol p.a. to

achieve a 50 μM concentration, resulting in a dark

purple solution (Furqan and Nurman, 2020). For

the preparation of the test solution, variations in

concentrations of 50 ppm, 30 ppm, and 10 ppm

are achieved by first weighing 50 mg of putat

extract and dissolving it in 10 mL of methanol p.a.

to create a 1000 ppm standard solution. Dilution

is then performed to obtain the desired

concentrations. The positive control solution is

prepared by dissolving 0.001 g of ascorbic acid in

5 mL of methanol p.a., forming a 100ppm

standard solution, from which further dilutions

are made to create solutions with concentrations

of 50 ppm, 30 ppm, and 10 ppm. The negative

control solution is composed of methanol p.a.

with the addition of DPPH solution. For the

antioxidant activity test, 0.2 mL of each test

solution, as well as the negative and positive

control solutions, are pipetted into separate vials

and combined with 3.8 mL of 50 μM DPPH

solution. After homogenizing the mixtures, they

are incubated in the dark for 30 minutes.

Absorbance is then measured at a wavelength of

517 nm using UV-Visible spectrophotometry. The

percentage of inhibition, which reflects the DPPH

scavenging activity, is calculated, along with the

Results and Discussions

The results of the data stated that in ethyl acetate

extract of putat leaves there are flavonoid

compounds, tannins and steroids. Method in

conducting phytochemical tests by looking at

color change reactions using color reagents. The

results of phytochemical tests on ethyl acetate

extract of putat leaves (Planchonia valida) can be

seen in Table 1.

Table 1. Secondary metabolites of Ethyl Acetate

Extract

Secondary Methabolite

Result

Alkaloids

Flavonoids

Tannins

Saponins

Steroids

-

+

-

+

Antioxidant of Crude Extract

The results of the antioxidant activity test of ethyl

acetate extract of putat leaves can be seen in

162

Chempublish Journal, 6(4) 2022, 160-167

Faction

Vial Bottle

Order

4

Fraction Weight

Table 2. Antioxidant activity of ethyl acetate

extract

(g)

1

2

3

4

0.050

0.056

0.060

0.050

Samples

Linear

Regression

y= 0.331x +

32.828

IC₅₀

(ppm)

Activity

5-8

9-15

16-18

Extract

Ethyl

Acetate

Ascorbic

acid

51.87

14.94

Strong

R2= 0.8421

Furthermore, qualitative antioxidant activity tests

were carried out on all fractions obtained to

determine the intensity of the resulting color

change. The qualitative test begins with the

elution of the fraction obtained using ethyl

methanol eluents in a ratio of 7 and 3 so that

Y

=

Very

Strong

2.34x

+ 14.943

R2 = 0.9731

The IC₅₀value of ethyl acetate extract of putat

leaves was obtained using the calculation of the

linear regression equation of ethyl acetate

extract of putat leaves in Figure 5 is y = 0.331x +

32.828 and R 2 = 0.8421. The y coefficient of this

equation expresses, while the x coefficient

expresses the amount of concentration required

to reduce 50% of DPPH radical activity. The linear

regression curve in Figure 1 illustrates that with

increasing concentration of extract, the greater

the % inhibition, which means the higher the

antioxidant activity. The following is a calculation

of the IC₅₀value of ethyl acetate extract of putat

leaves.

stains

are obtained on

the KLT

plate.

Furthermore, the KLT plate is sprayed using

DPPH solution to see the resulting color change

and presented through Figure 2.

Figure 2. Qualitative activity test using DPPH

sprayer against Ethyl Acetate Fraction

In Table 2, it was found that the IC₅₀value of 51.87

ppm is still in the strong range at the level of

antioxidant power according to Jun et al (2003),

due to it has an IC₅₀value in the range (50-100

ppm), so that ethyl acetate extract has the

potential to proceed to compound isolation at

the next stage

From Figure 2, it can be seen that the dominant

color intensity is found in fraction 1. In addition

to being based on the resulting stain pattern,

grouping fraction 1 is based on a less tailless stain

pattern, it can be possible that fraction 1 could

potentially contain pure compounds. Therefore,

recrystallization is carried out to remove the

Isolate of Planchonia valida

conyaminants

contained

in

the

sample.

A total of 15 grams of extract was impregnated

using 15 grams of silica gel. The stationary phase

used is 40 grams of silica gel. The sample was

eluted using ethyl acetate eluent: methanol in a

gradient, the polarity increased by 10%. From the

isolation process carried out, 22 vials were

accommodated based on the color of the

Purification is carried out by recrystallization

using solvents from nonpolar (n-hexane),

semipolar (ethyl acetate), polar (methanol) after

which it is dried.

Isolates

were

monitored

by

TLC

using

appropriate eluents (Methanol : ethyl acetate =

3:7) and stains were obtained. A single stain was

obtained on the KLT plate using Methanol eluent:

Ethyl Acetate = 3: 7. This identifies that the

isolates obtained are already pure. However, to

ensure that the monitor is carried out again by

KLT using a different eluent development (3

eluent system). The eluent used is chloroform ;

resulting tape. Furthermore,

a

Thin Layer

Chromatography (TLC) test is carried out by

observing the same stain pattern from each vial

to identify the combined fraction. The following

is a table of KVC result fraction grouping.

Table 3. Grouping of KVC yield fraction of ethyl acetate

extract of putat leaves

163

Chempublish Journal, 6(4) 2022, 160-167

ethyl acetate (1:9); DCM: Chloroform (2:8) and

DCM: n-hexane (1:9) and produced a dead stain.

This identifies that the compounds contained in

F1 isolates are pure (Figure 3).

Chemical Structure Determination

Characterization

using

a

UV-Vis

spectrophotometer to determine the presence of

chromophore groups contained in an organic

compound. The working principle of the UV-Vis

spectrophotometer is the interaction between

radiation in the range of 200-800nm wavelengths

passed to a compound. This interaction results in

transitions between electronic energy in organic

molecules. The following are the results of

measuring the wavelength of isolates using a UV-

Vis spectrophotometer (Figure 4).

Isolates

were

monitored

by

TLC

using

appropriate eluents (Methanol : ethyl acetate =

3:7) and stains were obtained. A single stain was

obtained on the KLT plate using Methanol eluent:

Ethyl Acetate = 3: 7. This identifies that the

isolates obtained are already pure. However, to

ensure that the monitor is carried out again by

KLT using a different eluent development (3

eluent system). The eluent used is chloroform ;

ethyl acetate (1:9); DCM: Chloroform (2:8) and

DCM: n-hexane (1:9) and produced a dead stain.

This identifies that the compounds contained in

F1 isolates are pure (Figure 3).

Figure 4. UV-Vis Spectrum of F1 Isolates Ethyl

Acetate 1λ = 235nm and 2λ = 286nm.

Figure 3. F1 isolate KLT results using eluents

(from left to right) DCM : ethyl acetate = 1:9,

chloroform : ethyl acetate = (1:9) and DCM :

Chloroform = (2:8).

The purpose of characterization by UV-Vis

spectrophotometer aims to analyze compounds

containing chromophore groups, which are part

of molecules that absorb UV light and visible

light. Figure 4 shows the maximum absorption at

wavelength. UV-Vis spectra show that isolates

provide two absorption peaks: in band 1 λ =

235nm and band 2 λ = 286nm. Absorption at 235

indicates the existence of a conjugated diema

system in the structure. Meanwhile, absorption

in 286 indicates the existence of an aromatic

system with certain substituents.

Phytochemical screening

Based on the table above, it can be seen that

ethyl acetate isolate gave positive results in

testing

Lieberman

steroid

class

compounds

reagents which

using

are

Burchard

characterized by the formation of green / blue

color in the sample. After obtaining preliminary

information related to the compounds contained

in the isolate, it is necessary to characterize

spectrophotometrically at the next stage.

Table 4. Phytochemical screening results of ethyl

acetate isolate

Secondary Metabolites

Alkaloids

Result

-

Tannins

Flavonoids

Saponins

Steroids

-

+

Figure 6. IR Spectrum of F1 Ethyl Acetate

164

Chempublish Journal, 6(4) 2022, 160-167

Identification of isolates by IR spectrophotometry

showed that absorption at wavenumbers

2920.76 cm-1 and 2851.25 cm-1 is suspected to

be absorption of the C-H group^.This conjecture is

strengthened by the presence of bending

vibrations in the wavenumber region 1462.83

cm-1 which indicates the presence of CH 2

groups. The absorption band in the wavenumber

region 13,78,30cm-1 shows the bending of the C-

H group of CH₃. The absorption band at

wavenumber 1734.83 cm-1 is thought to be an

absorption for the C=O group, as Hiroshi et al

(1976) reported, there is an O-H bond (hydroxyl)

and has a double bond C=C (ena) and a C=O bond

(carbonyl). The presence of O-H (hydroxyl) bonds,

having a C=C (ena) double bond and a C=O

(carbonyl) bond identifies that isolates of Fraction

1 Ethyl Acetate are Ergosterol compounds.

seen that the IC₅₀value of isolates is greater than

the IC₅₀value of ascorbate acid, which is 14.9488

ppm < 714.6377 ppm. A higher IC₅₀value of a

sample indicates weaker antioxidant activity.

Therefore, it can be concluded that the

compound demonstrates antioxidant activity

that is classified as weak, yet it still holds potential

as an antioxidant

Table 6. Antioxidant activity of Isolate

Samples

Linear

Regression

y= 0.0138x

+ 40.138

IC₅₀(ppm)

Isolate (F1)

714.63

R2 = 1

Ascorbic

acids

Y = 2.34x

+ 14.943

R2 = 0.9731

14.94

Conclusion

The ethyl acetate fraction of Putat Leaves

(Planchonia valida) contains a class of steroid

compounds. The antioxidant activity of extracts

and isolates of ethyl acetate fraction of Putat

Leaves (Planchonia valida) expressed in IC50

values is classified as strong and classified as very

weak

Figure 6. Chemical structure of Ergosterol

Table 5. Interpretation and Comparison of IR

Spectrum

Acknowledgement

Interpretation

Wavenumbers (cm^-1)

This research is supported by LPP Universitas

Isolate

2920.76

Ergosterol

2956.29

Jambi

through

Skema

Penelitian

Dasar

DIPA-

C-H

Penugasan

with

number:

SP

Alkanes/aromatics

C-H

O23.17.2.677565/2022 and Contract Number

272/UN21.11/PT.01.05/SPK/2022, July 2022.

2851.25

2871.43

Alkanes/aromatics

C=O

C=N, C=C

OH Bending

C-N aromatic

C-OH

1734.83

1462.11

1378.30

1244.29

1172.16

-

Author Contributions

1460.98

1369.64

1158.32

Conceptualization, N and DVS.; Methodology, FF

and RDP; Software, ILT and ML.; Validation: N and

ML; Formal Analysis, ILT and ML.; Investigation,

IIR and N.; Resources, N and FF.; Data Curation,

DVS and RDP; Writing

–

Original Draft

From the results of antioxidant activity tests in

Table 6 conducted on F1 isolates and ascorbate

acid, it is known that F1 isolates have an IC₅₀value

of 714.6377 ppm. This indicates that F1 isolates

have weak antioxidant levels (Jun et al, 2003). This

is reinforced by comparing the IC₅₀value of

isolates with the IC₅₀value of ascorbate, it can be

Preparation, DVS, N; Writing – Review & Editing,

ILT and ML; Visualization: N and ML.; Supervision,

N and ML; Project Administration, ML

Conflic of Interest

The authors declare no conflict of interest

165

Chempublish Journal, 6(4) 2022, 160-167

References

Pratiwi, D. I., R. A. Syarif., R. Waris dan Faradiba.

2019. “Isolasi Senyawa Antioksidan Ekstrak

Metanol Kulit Buah Naga Merah (Hylocereus

polyrhizus). Jurnal Fitofarmaka Indonesia. Vol. 6

(1) : 340-346.

Shao, S.,M. Hernandez., K. G. john., Kramer., L.

Danny dan R. Tsao. 2010. Ergosterol Profiles,

Fatty Acid Composition, and Antioxidant

Activities of Button Mushrooms as Affected by

Tisuue Part and Develpmental Stage. Food

Chem. Vol. 58 (22) : 11616-11625.

Shaumi, N. 2019. Karakterisasi Spesifik dan Non

Spesifik Simplisia Serbuk Pucuk Daun Putat

(Planchonia valida). Skripsi. Sekolah Tinggi Ilmu

Kesehatan : Samarinda.

Supriningrum, R., Fatimah, N., dan Purwanti, Y. E.

2019. Karakterisasi spesifik dan non spesifik

ekstrak etanol daun putat (Planchonia valida).

AI Ulum sains dan Teknologi. Vol 5 (1) : 6-12.

Suryelita., S. B. Etika dan N. S. Kurnia. 2017.

Isolasi Dan Karakterisasi Senyawa Steroid Dari

Daun Cemara Natal (Cupressus funebris Endl.).

Vol. 18 (1)

Ardji, A. Widiyantoro dan L. Destiarti. 2018.

Karakterisasi Senyawa Steroid Dari Fraksi

Diklorometana Batang Tanaman Andong

(Cordyline

Fruticosa)

dan

Aktivitas

Sitoktosiknya Terhadap Sel Hela. JKK. Vol. 7(1)

:48-52.

Furqan, M., dan Nurman, S. 2020. Ekstrak polar

Kopi hijau arabika sebagai antihiperglikemi

pada mencit (Mus Musculus). Journal of

Healthcare Technology and Medicine,Vol. 6 (2)

: 1323-1331.

Iglesias,

C.G ^., E. Salas ^., N. Barouh ^., B. Baréa,

Panya ^., M.

C. Figueroa-Espinoza.

2016.

Antioxidant activity of protocatechuates

evaluated by DPPH, ORAC, and CAT methods.

Vol. 194 : 749-757

Ipandi, I., L. Triyasmono dan Prayitno. 2016.

Penentuan

kadar

Flavonoid

Total

dan

Aktivitas Antioksidan Ekstrak Etanol Daun

Kajajahi (Leucosyke capitellata Wedd.). Jurnal

Pharmascience. Vol. 3(10 : 93-100.

Handayani, S., I. Kurniawati dan F. A. rasyid. 2020.

Syamsudin, D., A. H. Alimuddin dan B. Sitorus.

Isolasi Dan Karakterisasi Senyawa Fenolik Dari

Daun Putat (Planchonia valida Blume). 2019.

Indonesian Journal of Pure and Applied

Chemistry. Vol. 5 (2) : 85-98.

Syukur, M. 2016. Habitat Pohon (Barringtonia

acutangular) pada Kawasan Berhutan Sungai

Jemelak Kabupaten Sintang. PIPER. 12 (23) : 74-

82.

Uji Aktivitas Antioksidan Ekstrak Daun Karet

Kebo

(Ficus

elastica)

dengan

Metode

Peredaman Radikal Bebas DPPH (1,1-Diphenyl-

2-Picrydazil). Journal of Pharmacy. Vol 6910 :

141-150.

Jamiat., Iskandar dan M. Idham. 2019. KEARIFAN

LOKAL MASYARAKAT DALAM MELESTARIKAN

LEBAH MADU ALAM DENGAN TEKNIK TIKUNG

DI KAWASAN SIAWAN BELIDA KAPUAS HULU.

JURNAL HUTAN LESTARI. Vol. 7 (2) : 743-752.

Leksono, W.B., Pramesti, R., Santosa, G.W., &

Setyati,

Metanol Dan N-Heksana Terhadap Aktivitas

Antioksidan Ekstrak Rumput

Laut Gelidium sp.. 21(1), 9-16

Manae, M., Gheevarghese,R dan Chiosa, V. 2007.

MolecularInteraction Of ACholineterase

Activator With Studid By

HighResolution Spectrometry.

Vanessa, M., R. L. Munhoza., R. P. José., A. C.

João., E. Zequic., M. Leite., C. Gisely., J. P.

Lopesa dan Melloa.

(2014). Extraction Of

Flavonoids From Tagetes Patula: Process

Optimization And Screening For Biological

Activity. Rev Bras Farmacogn Vol 24 : 576-583.

Wardana, A.P., dan Tukiran. 2016. Skrining

fitokimia dan aktivitas antioksidan ekstrak

W.A.

(2018).

Jenis

Pelarut

kloroform

polycephalum). Prosiding Semnas Kimia dan

Pembelajarannya, Jurusan Kimia FMIPA

Universitas Negeri Surabaya: 4.

tumbuhan

gowok

(Syzygium

Ergosterol

FTIR

Department of Physical Chemistry. 2(4) : 33-

37.

Ningrum, R., Elly, P., dan Sukarsono. 2016.

Identifikasi Senyawa Alkaloid Dari Batang

Wahdaningsih, S., S. Wahyuono dan E. P.

Setyowati. 2013. Isolaso dan Identifikasi

Senyawa Antioksidan Daun Pakis (alsophilia

glauca J.SM) dengan Metode DPPH (2-2-

difenil-1 pikrihidazil )”. Tradisional Medichine

Journal. Vol. 18 (1): 38-45.

Karamunting

Sebagai Bahan Ajar Biologi Untuk SMA kelas X.

Jurnal Pendidikan Biologi Indonesia. 2(3): 231.

(Rhodomyrtus

tomentosa)

166

Chempublish Journal, 6(4) 2022, 160-167

Wulansari, A. N. 2018. “Alternatif Cantigen Ungu

(Vaccinium varingiaefolium) Sebagai

Antioksidan Alami: Review”. Farmaka. Vol 6(2):

419-429

167