The DPPH Radical Scavenging Activity of Freeze-dried and Spray-

dried Soluble Coffee Brew

I Dewa Ayu Susilawati1*), Fredy Akbar2), Didin Erma Endahyani3)
1, 2, 3
Faculty of Dentistry Universitas Jember, Indonesia
*Corresponding Author: dewasusi@unej.ac.id

Abstract
The antioxidant activity of soluble (instant) coffee brew was supposed to be different
from the regular coffee powder brew. It might be due to the different coffee processing that
affect to its chemical composition. This study aimed to analyze the antioxidant activity of
instant Robusta and Arabica coffee brew processed by freeze dried and spray dried.
Antioxidant activity was analyzed using DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate)
radical scavenging method and expressed as the percentage of DPPH radical inhibition.
This study examined six type samples i.e. 1) spray dried Robusta instant coffee (SDR), 2)
freeze dried Robusta instant coffee (FDR), 3) regular Robusta coffee powder (RR), 4) spray
dried Arabica instant coffee (SDA), 5) freeze dried Arabica instant coffee (FDA), and 6)
regular Arabica coffee powder (RA). Regular Robusta and Arabica coffee powder were
purchased from the same fabric and used as controls. Data were analyzed using Anova and
LSD. Result showed that instant coffee brew (both Robusta and Arabica) processed by
freeze drying and spray drying demonstrated a significantly higher (p<0,05) DPPH radical
scavenger activity compared to regular coffee powder brew. Freeze-dried Arabica instant
coffee brew showed the highest (p<0,05) antioxidant activity. Further studies were needed
to elucidate the composition of antioxidants components of different kinds coffee samples
from different processing.

Key words: Antioxidant; Arabica; Robusta; Coffee processing

INTRODUCTION

Coffee brew is one of the healthy drinks. Beside well known to boosts the mental alertness
and cognitive functions, it’s rich of antioxidants (Yashin et al, 2013) that can counteract to
the oxidants and free radicals, and therefore might prevent to oxidative stress and diseases.
Coffee was reported to reduce the production of neutrophils superoxide radical in vitro
[Susilawati et. al. 2019], study using atherosclerotic rat model showed that coffee
consumption protected cardiovascular health [Susilawati et.al. 2020]. ……

Coffee brew can be served from the regular or instant ground coffee. Regular ground coffee
brew is usually reminding the spent after brewing, while instant coffee is a totally soluble
coffee without reminding the spent. Instant coffee is a beverage that enables people to
quickly prepare hot coffee by adding hot water to the powder and stirring manually.

Instant coffee is commercially prepared by either freeze-dried or spray-dried processing

(Burmester, 2011). The distinction of coffee processing might cause the change in the
chemical composition and affect to the antioxidant activity (Jeong et al 2013; Odžaković et
al, 2016; Marcucci et.al. 2017). There was only a little study concern with the health benefit of
instant coffee. This study aimed to analyze the antioxidant activity instant coffee brew and
compare to the regular coffee brew

MATERIAL AND METHODS

There were six type samples in this study: 1) spray dried Robusta instant coffee (SDR), 2)
freeze dried Robusta instant coffee (FDR), 3) regular Robusta coffee powder (RR), 4) spray
dried Arabica instant coffee (SDA), 5) freeze dried Arabica instant coffee (FDA), and 6)
regular Arabica coffee powder (RA). The commercial regular coffee powder (Arabica and
Robusta) were purchased from the same fabric i.e. State Coffee Plantation PTPN XII
Indonesia. Coffee spray drying and freeze drying were processed in Agriculture
Technology Laboratory Faculty of Agricultural Technology, Universitas Jember Indonesis,
using equipment Vacuum freeze dryer (Toption TPV-50F) and LabPlant SD-06 Spray
Dryer. Reagent for DPPH (1,1-diphenyl 2- picrihydrazyl) was purchased from Sigma

Preparation of spray dried instant coffee

Spray dried instant coffee, in this study, was produced from the commercial regular coffee
powder. Basically, the extract of regular coffee powder was sprayed into hot air to form tiny
droplets. Then, the heat dries and turns them into a fine powder or small pieces (modification
method by Ianniciello et al, 2014). A total of 250 g regular coffee powder (Robusta or Arabica)
was brewed in 2 L boiled distillated water, stirred using ultrasonic stirrer for five minutes, and
then filtered (100 Mesh). The extract was sprayed into spray-dried apparatus, at the
temperature inlet 120oC and outlet 60oC. The fine powder was then collected and preserved
in the sterile plastic bag and keep at the room temperature until before being analyzed. This
process was yielding two kinds of samples i.e. spray dried Robusta instant coffee (SDR)
and spray dried Arabica instant coffee (SDA)

Preparation of freeze dried instant coffee

Freeze dried instant coffee was prepared from commercial regular coffee powder (same
sample for making SD instant coffee). Principally, the extract of regular coffee powder was
frozen and then dried at a low temperature in a vacuum. A total of 250 g regular coffee powder
(Robusta or Arabica) was brewed in 2 L boiled distillated water, stirred using ultrasonic stirrer
for five minutes, and then filtered (100 Mesh). The supernatant was then frozen at minus
temperature (-50oC ) for 48 h until forming fine powder. This procedure was yielding
samples freeze dried Robusta instant coffee (FDR) and freeze dried Arabica instant coffee
(FDA)
Metode pengukuran aktivitas antioksidan pada kopi menggunakan metode DPPH
Metode yang sering digunakan untuk mengukur aktivitas antioksidan salah satunya
adalah menggunakan radikal bebas DPPH (1,1-diphenyl-2-picrihydrazyl). DPPH
merupakan radikal bebas yang stabil pada suhu kamar dan sering digunakan untuk menilai
aktivitas antioksidan dari beberapa senyawa bahan alam. Metode ini dipilih akrena mudah
cepat, sederhana dan peka untuk menilai aktivitas antioksida dari bahan alam. Pada metode
DPPH adalah aktivitas penghambatan radikal bebas yang merupakan interaksi bahan
antioksidan dengan DPPH secara transfer elektron maupun radikal hydrogen dan akan
menetralkan karakter radikal bebas DPPH. Setelah larutan sampel dicampur dengan DPPH,
aktivitas peredaman dapat ditandai dengan adanya perubahan warna dari ungu, ungu pudar,
hingga kuning.

Uji antioksidan seduhan bubuk kopi

Penetapan konsentrasi ppm bagaimana ?
sampel masing masing dipreparasi dengan cara sampel dilarutkan menggunakan
aquades dengan konsentrasi 100 ppm, 250 ppm, 500ppm, 750 ppm, 1000 ppm, dan 1500.
Sampel kemudian difilter menggunakan kertas saring. Filtrat yang diperoleh kemudian diuji
aktivitas antioksidan menggunakan metode DPPH. Filtrat masing masing konsentrasi
kemudian dimasukkan dalam kuvet dengan perbandingan 100 µl larutan induk kopi, 400 µl
etanol, dan 500 µl DPPH. Kemudian, di inkubasi selama 30 menit pada suhu ruangan
dengan kondisi tertutup hingga terjadi perubahan warna dari aktivitas DPPH. Semua
sampel yaitu sampel ekstrak yang telah di inkubasi di uji nilai absorbansinya menggunakan
spektrofotometer Uv-vis pada panjang gelombang 514 nm.

Penentuan % inhibisi
Untuk mencari % inhibisi dapat dihitung menggunakan rumus sebagai berikut
|blanko|−|sampel|
% inhibisi = x 100 %
|blanko|

HASIL
Pengujian aktivitas antioksidan dilakukan untuk mengetahui % antioksidan sampel.
Dari hasil pengujian, % inhibisi dari sampel dapat dilihat pada tabel 1.

Tabel 1. Coffee scavenging activity against DPPH radical

Coffee scavenging activity against DPPH radical (Percentage of inhibition)
RB RS RF AB AS AF
100 ppm 1.69±1.50 8.32±1.10 29.77±0.71 7.36±0.34 15.36±3.93 16.18±0.37
250 ppm 13.85±1.58 34.75±2.36 66.71±0.65 26.84±0.87 38.91±1.27 59.67±0.92
33.88±0.8 87.19±1.9 69.63±6.0
500 ppm
3 61.22±0.89 6 54.36±1.76 9 82.76±1.16
54.36±0.5 83.50±0.1 85.09±1.5
750 ppm
5 78.09±2.23 5 72.19±1.24 8 84.46±0.37
72.51±1.9 84.72±1.0 82.71±1.8
1000 ppm
5 77.22±1.48 5 83.08±0.84 1 84.07±0.52
 82.99±3.3 84.72±2.0
1500 ppm
3 85.32±1.07 83.81±2.21 83.08±0.84 0 84.46±0.30

RR: regular Robusta coffee powder RA: regular Arabica coffee powder
SDR: spray dried Robusta instant coffee SDA: spray dried Arabica instant coffee
FDR: freeze dried Robusta instant coffee FDA: freeze dried Arabica instant coffee
 100
90
80
persen inhibisi (%) 70
r
60 b
50 rs
rf
40
a
30 b
20 a
s
10 af
0
100 250 500 750 1000 1500
konsentrasi (ppm)

RR: regular Robusta coffee powder

SDR: spray dried Robusta instant coffee
FDR: freeze dried Robusta instant coffee
RA: regular Arabica coffee powder
SDA: spray dried Arabica instant coffee
FDA: freeze dried Arabica instant coffee

Figure 1. Coffee scavenging activity against DPPH radical

Discussion
Conclusion
Aknowledgement

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