Rapid evaluation and comparison of natural products

and antioxidant activity in calendula, feverfew, and

German chamomile extracts

19/447943/PFA/0194 Fella
8 Syifa
 LATAR BELAKANG

Feverfew (Tanacetum
parthenium) termasuk dalam
keluarga Asteraceae dan
secara empiris digunakan
untuk pengobatan migrain
Parthenolide, suatu
seskuiterpen lakton,
merupakan senyawa aktif
dalam daun feverfew
Parthenolide dapat detemukan
dalam spesies Asteraceae lain
seperti Chamomile German
(Matricaria recutita)
Migrain kemungkinan
disebabkan karena
terbentukanya radikal bebas
dalam otak dan menyebabkan
kerusakan oksidatif
Adanya senyawa antioksidan
dalam fevervew mungkin
berkontribusi dalam
pengobatan migrain.
 TUJUAN

Membandingkan fingerprint
feverview, German
Chamomile, dan marigold
dengan HPTLC

Menentukan jumlah
kandungan apigenin,
chamazulene, bisabolol dan
parthenolide dalam ekstrak
daun dan kepala bunga
ketiga sampel

Mengembangkan metode
standarisasi RP HPTLC-DPPH*
untuk mengukur dan
membandingkan aktivitas
antioksidan antar ekstrak
dan antar senyawa aktif
 BAHAN DAN METODE
Bahan
German chamomile, feverview
dan marigold ditanam selama
5 bulan pada suhu konstan
21oC dengan cahaya 12h/hari
Standaridised supercritical
CO2 extract
Standar: asam galat, apigenin,
chamazulene, bisabolol,
parthenolide, quercetin, rutin
Bahan HPTLC
Solven
Larutan Standar
0.1 mg/ml tiap larutan standar
dipreparasi dengan dilarutkan
dalam etanol absolut dan
disimpan dalam 4oC
Larutan ekstrak
supercritical CO2
50 mg/ml tiap larutan ekstrak
dipreparasi dengan dilarutkan
dalam kloroform dan
disimpan dalam 4oC
 BAHAN DAN METODE
Ekstraksi
10 g daun segar dipetik dan
dibekukan. Daun beku
dihancurkan dengan mortar
dan stamper, kemudian
dimasukkan dalam thimble
dan di refluks dalam soxhlet
selama 12h menggunakan
100ml etanol absolut.
Sari dipekatkan dengan rotary
evaporator hingga 10 ml,
dimasukkan dalam labu ukur
25ml add etanol absolut.
Simpan dalam 4oC
HPTLC
Plate fase normal HPTLC
dilakukan pre-wash sebelum
blank run dengan etanol,
dikeringkan, aktivasi di oven
pada 110oC 15 menit. Sampel
dispray 8mm menggunakan
Linomat 5.
Larutan untuk derivatisasi post-
kromatogram
Tiap plate di-spray menggunakan
larutan 0,4% w/v DPPH
 BAHAN DAN METODE
HPTLC
Plate kromatogram
dikembangkan dalam
chamber AMD 2 (Automated
Multiple Development
Chamber)
Fase gerak normal phase
heksan: EA: AA (20:10:1)
Fase gerak reverse phase
Aseton: air (90:10)
Analisis plate menggunakan
TLC-visualizer dibawah UV
366 nm dan 256 nm, dan
cahaya tampak
HPTLC
Plate yang telah
dikembangkan di foto
sebelum dan setelah
derivatisasi dengan
menyemprotkan larutan
anisaldehid (0.5:50:1) atau
0.4% DPPH.
HPTLC kuantitatif dianalisis
menggunakan VideoScan
Digital Image Evaluation
software
 BAHAN DAN METODE

Validasi metode

Kuantifikasi metode HPTLC Variasi antar replikasi

untuk apigenin, bisabolol, diekspresikan dalam %RSD
chamazulene dan
parthenolide dinilai dengan Ditentukan pula LOD dan
plotting peak area vs jml LOQ
sampel dengan range
tertentu.

Linieritas pada HPTLC-DPPH*

fase normal dan RP dievaluasi
pada standar asam galat.

Repeatability (presisi) dinilai

dengan dilakukan replikasi
pada 2 konsentrasi standar yg
berbeda
HASIL DAN PEMBAHASAN
 HPTLC fingerprint
visible 254 nm
chamazulen
e
bisabolol

parthenolide

apigenin

1. Chamomile supercritical flower head 7. Parthenolide and bisabolol;

extract; 8. Chamazulene;
2. Chamomile Soxhlet leaf extract; 9. Apigenin (light blue), parthenolide and
3. Feverfew supercritical plant extract; bisabolol
4. Feverfew Soxhlet leaf extract;
5. Marigold supercritical flower extract;
Validasi metode HPTLC
 Kuantifikasi HPTLC

• Apigenin hanya bisa terkuantifikasi pada

chamomile.
• Chamazulene dan bisabolol tertinggi
terdapat pada chamomile dan calendula
(flower head).
• Parthenolide tertinggi terdapat pada
Parhenolide dan chamazulene merupakan suatu seskuiterpene yang terdiri
dari 3 unit isoprene. Dalam tanaman, kedua senyawa tsb terbentuk dari
perkusor seskuiterpen yaitu farnesyl diphosphate (FPP).

ada 2 kemungkinan siklisasi FPP yaitu membentuk parthenolide dan

chamazulene (hasil degradasi dari proazulenic seskuiterpen lacton seperti
e.g matricin).
 HPTLC-DPPH*

Pemisahan senyawa menggunakan HPTLC kemudian plate di semprot dengan

larutan DPPH untuk deteksi aktivitas antioksidan

Dibandingkan HPTLC normal phase dan reverse phase

RP HPTLC lebih direkomendasikan karena tidak ada pengaruh dari reaksi senyawa
radikal dan antioksidan
Pada normal phase terjadi reaksi antara muatan positif silica gel dengan senyawa
polifenol, sehingga akan menguatkan reaksi dengan DPPH

DPPH merupakan radikal bebas stabil yang berwarna ungu gelap dan akan berubah
menjadi kuning pucat ketika direduksi oleh senyawa antioksidan dalam sampel.
Elektron bebas pada atom nitrogen akan direduksi dengan menerima atom hidrogen
membentuk hydrazine.
 HPTLC-DPPH*

Aktivitas antioksidan ditentukan dengan membandingkan area dan intensitas warna dari
pita kuning yang terbentuk setelah disemprot DPPH dengan kurva kalibrasi asam
galat.
Akurasi dan presisi pada RP HPTLC lebih tinggi

Akurasi pada normal phase baik kecuali pada konsentrasi 1.0 µg

01. OVERVIEW
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02. FEATURES OF THE TOPIC

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03. ASSIGNMENT
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04. ABOUT THE TOPIC

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1.
Despite being red, Mars is
actually a cold place. It’s full of
iron oxide dust, which gives the
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2.
Venus has a beautiful name and
is the second planet from the
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MARS VENUS
Despite being red, Mars is Venus is the second
actually a cold place planet from the Sun

SATURN MERCURY
Saturn is composed Mercury is the smallest
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helium
 FEATURES OF THE TOPIC

Jupiter is a gas giant Saturn is a gas giant, Despite being red,

and the biggest composed mostly of Mars is actually a
planet in the Solar hydrogen and helium cold place full of iron
System oxide dust
 PLANTS IN THE WORLD

6,500,000
Total amount of land plants
in the world

To modify this graph, click on it, follow

2,200,000
the link, change the data and paste Total amount of water plants
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 8.7 M
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● Venus has a beautiful name,

but it’s terribly hot

● Mercury is the closest planet

to the Sun

● Despite being red, Mars is

actually a cold place
DID YOU KNOW THIS?

Mercury is the closest

planet to the Sun

Venus is the second

planet from the Sun
Mercury is the closest planet
to the Sun and the smallest
one in the Solar System—it’s
only a bit larger than the
Moon

PROBLEM

SOLUTION

Venus has a beautiful name

and is the second planet from
the Sun. It’s terribly hot, even
hotter than Mercury
 THIS IS A TABLE

01 02 03
Saturn is composed of Neptune is the farthest Venus is the second
hydrogen and helium planet from the Sun planet from the Sun

Jupiter is the biggest

Despite being red, Mars Mercury is the closest
planet in the Solar
is actually a cold place planet to the Sun
System
AWESOME WORDS
 PROCESS

Jupiter is the
Venus is the
01 second planet
03 biggest planet in
the Solar
from the Sun
System

Despite being
Neptune is the
red, Mars is
actually a cold 02 farthest planet 04
from the Sun
place
 EXERCISE

1. Despite being red ____, is 2. Mercury is the smallest 3. Saturn is a ___ giant and
actually a cold place planet in the _____ ______ has several rings

4. _____ is the second planet 5. Neptune is the ________ 6. Jupiter is the _______ planet
from the Sun planet from the Sun in the Solar System
 OVERVIEW DIAGRAM

Botanical Method

Herbarium Botanical Garden ICN

Despite being red, Jupiter is the Saturn is composed

Mars is actually a biggest planet in mostly of hydrogen
cold place the Solar System and helium
 ASSIGNMENT

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Venus has a beautiful name, but
is its atmosphere poisonous?

2.
Does Mercury’s name has
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metal? B.

3.
What gives Mars its distinctive
C.
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