The Level of Flavonoid and the Antioxidant Activity of the Growol

Flour

Yasinta Nofia

, Brian Wasita

1,2

and Tri Nugraha Susilawati

Clinical Nutrition, Graduate School, Universitas Sebelas Maret, Surakarta, Indonesia

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

Department of Microbiology/Research Ethics Committee, Faculty of Medicine, Universitas Sebelas Maret, Surakarta,

Indonesia

Keywords: Growol Flour, Flavonoid, Antioxidant Activity

Abstract: Growol a fermented product made from cassava, has the potential as a functional food. The food is origin

from the Kulon Progo area, Yogyakarta. Growol flour contains high dietary fiber and can be consumed as

alternative food for people with Diabetes Mellitus (DM). This study aims to determine the concentration of

flavonoid and the antioxidant activity of growol flour. The flavonoid levels were determined by using the

FeCl

method followed by a spectrophotometry assay. The antioxidant activity of growol was measured by

using the 2,2-Diphenyl Picrylhydrazyl (DPPH) method. The assays showed that the average level of growol’s

flavonoid was 6.8004 mg/100g while the average antioxidant activity showed that inhibition percentage was

37.232%. Since growol flour contains flavonoids and have antioxidant activity, the food has the potential to

be consumed as an optional diet therapy for people with diabetes.

1 INTRODUCTION

Cassava is originating from the Americas. The plant

also called wood potato or kasepe and Manihot

Esculenta Crantz in Latin. Cassava contains

flavonoids, alkaloids, saponins, phenolics, and

glycosides (Hasanah et al., 2020; Yi et al., 2011). It is

known as a source of staple food after rice and corn

and is one of the plants that contain cyanogenic.

Despite its well-known "cyanogenic" content that

indicated its toxicity, processing cassava by

fermentation method is a way to reduce these toxic

components as is true for other plant cyanogenic

glycosides (Pramudita et al., 2017; Wu et al., 2012).

Growol is one of the popular fermented foods made

from cassava and considered safe.

Growol is a product of fermented cassava from

Kulon Progo, Yogyakarta, that has the potential as a

functional food. The growol fermentation goes

through an immersion stage for 3 days which can

reduce the level of cyanogenic toxicity so that it is at

a safe level. In addition, during this process,

fermentation by probiotic lactic acid bacteria (LAB)

occurs. LAB is gram-positive and beneficial for

human health including stimulating the immune

system, balancing intestinal flora, reducing the risk of

tumors, improving blood lipids, premature aging, and

its function in antioxidant activity (Ayeni et al., 2011;

Kuda et al., 2010; Lee et al., 2010). Fermentation of

foods can also increase antioxidant activity by

increasing the release of flavonoids from plant foods,

making it a useful method to increase the supply of

natural antioxidants (Hur et al., 2014).

Flavonoids are bioactive compounds that can act

as antioxidants. Antioxidant compounds can inhibit

the entry of free radicals into the human body by

giving electrons to free radical molecules so that these

molecules become stable (Hasim et al., 2016). To our

knowledge, the flavonoid levels and the antioxidant

activity in growol has not been well studied. In

previous studies, only research related to the analysis

of prebiotics contained in growol flour was carried

out (Puspaningtyas et al., 2019). Thus, this study aims

to determine the flavonoid levels and the antioxidant

activity of growol flour.

2 METHODS

This research was conducted at the Central

Laboratory of Food and Nutrition Studies (PSPG) in

Noﬁa, Y., Wasita, B. and Susilawati, T.

The Level of Flavonoid and the Antioxidant Activity of the Growol Flour.

DOI: 10.5220/0010759100003235

In Proceedings of the 3rd International Conference on Social Determinants of Health (ICSDH 2021), pages 257-260

ISBN: 978-989-758-542-5

257

June 2021. The type of research in this study was

experimental laboratory in vitro to determine the

levels of flavonoids and the antioxidant activity in

growol flour.

2.1 Instrument and Material

The instrument used in this research are knife,

container, steam pot, cabinet dryer, grinder, 60 mesh

sieve, test tube, scale, UV-Vis spectrophotometer.

The main ingredients used in the production of

growol flour are cassava, ethanol 96% and methanol.

2.2 Growol Production

The process of making growol is carried out with

modifications and refers to previous research

(Puspaningtyas et al., 2019).



Figure 1: The process of making growol

2.3 Growol Flour Production

Growol flour manufacture refers to previous research

(Puspaningtyas et al., 2019).



Figure 2: The process of growol flout poduction

2.4 Measuring Flavonoid Levels

About of 1-2 grams of growol flour was weighed and

put in a test tube, and then added with 10 ml of 96%

ethanol. Next, 1 ml of mother liquor and add 5 ml of

FeCl3 solution (forming a red color) was added. It

was then diluted to 10 ml using 96% ethanol. UV-Vis

spectrophotometer was used for tera with a

wavelength of 520 nm.

2.5 Measuring Antioxidant Activity

2.5.1 Antioxidant Activity/Radical

Scavenging Activity

Antioxidant Activity/ Radical Scavenging Activity (RSA)

is conducted refer to previous research (Yen & Cheng,

1995)

. About 1-2 g of growol flour was weighed and

dissolved using methanol at a certain concentration.

Then, 1 ml of mother liquor was taken and put in a

test tube. Next, 1 ml of 1,1, 2,2–Diphenyl Picryl

Hydrazyl (DPPH) solution, 200 micromolars, was

added. Then, it was incubated in a dark room for 30

minutes. It was diluted to 5ml using methanol. A

blank was made by adding 1 ml of DPPH solution

into 4 ml of ethanol. The tera was at a 515 nm

wavelength.

% Inhibition =

  



x100%

(1)

3 RESULTS AND DISCUSSION

Growol is a fermented food typical of Kulon Progo

made from cassava through a process of soaking,

Sortin

The clean cassava is washed

with running water

Soaking cassava for 3 days

Cassava is washed again and

filtered (4 times)

Cassava is squeezed and dried

Milling

Raw Growol

Raw growol steaming

Growol

Dried in a cabinet dryer at 80

± 6 hours

Floured using a grinder

Growol Four

Sieved through a 60-mesh sieve

ICSDH 2021 - International Conference on Social Determinants of Health

258

washing, grinding, and steaming to produce mature

growol (Puspaningtyas et al., 2019).

The levels of total flavonoid in growol flour were

determined by using UV-Vis spectrophotometer with

calorimetric method. The use of a spectrophotometer

in determining flavonoids is due to the presence of a

conjugated aromatic system in flavonoids which can

show strong absorption bands in the ultraviolet and

visible spectrum regions. The maximum wavelength

used is 520 nm.

FeCl3 is used for testing the levels of flavonoid in

growol flour, followed by UV-Vis spectrophotometer.

Flavonoids are one of the natural compounds that can

act as antioxidants and hypolipidemics (Nwose et al.,

2017). They can also act as antidiabetic including

improvements in glycolysis pathways, mitochondrial

function, improving insulin sensitivity, reducing

oxidative stress and gluconeogenesis (Vinayagam

and Xu, 2015). Nwose et al., (2017) state that

flavonoid in cassava has anti-inflammatory,

hypolipidemic, and management effects on metabolic

syndrome disease. The levels of flavonoid are

presented Table 1.

Table 1: Flavonoid Levels in Growol Flour.

Sample

Results

(mg/100g)

Average

(mg/100g)

Growol Flour

6.7317

6.8004

6.8691

The flavonoids in growol flour is the result of the

fermentation process in growol. During the

fermentation, there was an increase in flavonoids due

to the presence of sugar-breaking enzymes and

degraded phenolic complex compounds because of

lactic acid bacteria activity. These enzymes support

the formation of flavonoid compounds due to the

addition of phenol groups

(Dwiputri, 2018).

These studies have shown that regular flavonoid

consumption of 5-100 mg/kg by oral or

intraperitoneal injection can provide an

antihyperglycemic effects (Ghorbani Ahmad, 2017).

In experimental animal studies with type 2 diabetes

mellitus, regular flavonoid consumption of 5-100

mg/kg can decreased fasting blood glucose and non-

fasting blood glucose (A. Hunyadi et al., 2012; C.Y.

Hsu et al., 2014; R. Jadhav, 2012). The mechanism of

regular flavonoid consumption on the

antihyperglycemic effect is the reduced absorption of

glucose from the small intestine by inhibiting

glucosidase and α-amylase involved in carbohydrate

digestion (O.M. Ahmed et al., 2010; R. Jadhav, 2012;

S. Jo, E. Ka et al., 2009; Y.Q. Li et al., 2009). Our

study showed that the levels of total flavonoid in

growol flour is 6.8004 mg/100 g. The application of

growol flour with a concentration of 6.8004 mg/100g

can also able to provide antihyperglycemic treatment.

Antioxidant activity is influenced by active

compounds contained in foods such as flavonoids

(Dewi, S. R. et al., 2018). The antioxidant activity of

growol flour is measured using the 2,2-Diphenyl

Picryl Hydrazyl (DPPH) method. DPPH is a method

of detecting antioxidant activity in a simple, fast, and

inexpensive way, and requires only a small number of

samples. This method is a stable and free radical at

room temperature and is very soluble in methanol and

is very easily oxidized by temperature and air. In

addition, DPPH is in powder form and is purple black

in color (Molyneux, 2004). The results of antioxidant

activity test presented in Table 2

The antioxidant activity in fermented food can

inhibit the entry of free radicals into the body and

improve the levels of glucose and lipid in the blood

(Bajaj & Khan, 2012; Ceriello & Testa, 2009; Li et

al., 2012). The result of antioxidant activity test

showed that inhibition percentage found in growol

flour is 37.2317% due to the presence of lactic acid

bacteria formed during immersion and the

fermentation process (Li et al., 2012; Nugraheni,

2011; Putri et al., 2012). The antioxidant activity is

due to the presence of flavonoids in growol flour.

Table 2: Antioxidant Activity in Growol Flour.

Sample Results (%) Average (%)

Growol Flour

37.1244

37.2317

37.3390

4 CONCLUSIONS

The levels of flavonoid and the antioxidant activity

with percentage inhibition in growol flour were

6.8004 mg/100g

and 37.2317% influenced by LAB

during the fermentation process in making the growol.

The levels of flavonoids and the antioxidant activity

in growol flour can be an alternative prevention or

diet therapy for people with diabetes mellitus.

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