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Free radical scavenging capacity, antibacterial activity and
essential oil composition of turmeric (Curcuma domestica)
varieties grown in Ethiopia

Belay Haile Kebede, Sirawdink Fikreyesus Forsido, Yetenayet B. Tola, Tessema Astatkie

By: Belay Haile

11th Annual Research Conference of Jimma University and 10th Global Knowledge Exchange
Network, April 2021. 

Jimmaa
Ethiopia

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Presentation Out lines

 Introduction

 Objectives

 Material and Methods

 Statistical Analysis

 Result and Discussion 2

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Introduction
Humankind is often vulnerable to oxidative stress due to ROS
and RNS as well as to diseases caused by bacterial infections
(Niamsa and Sittiwet, 2009; Tanvir et al., 2017).

Peoples take antioxidants and antimicrobials in a form of

commercial food additives that are produced synthetically to
counter the effects of oxidative stress and growth of food
spoiling microbes (Saǧ dıç and Ozcan, 2003; Akter et al., 2019).

Due to consumer health concerns related to the safety of food

containing synthetic chemicals, there is a growing interest in
using natural antioxidants and antibacterial compounds (Norajit
et al., 2007; Vallverdú -Queralt et al., 2015).
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… Introduction
Turmeric is a known spice indispensable for food
preparation and is reported to possess different chemical
properties and biological activities.

EOs and diarylheptanoid /curcuminoids are the major

secondary metabolites of turmeric that are used in a wide
range of pharmacological applications (Ravindran et al.,
2007; Prasad and Aggarwal, 2011).

 These active principles of turmeric rhizomes show a wide

range of biological activities such as antibacterial,
antifungal, anticancer, insect repellant and anti-snake
venom activity (Negi et al.,1999; Kocaadam and S¸ anlier,
2017; Dosoky and Setzer, 2018).
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… Introduction
However, the amount of bioactive extracts, antioxidant
and antimicrobial properties depend on the type of
chemical composition and cultivated turmeric
varieties (Tanvir et al., 2017; Akter et al., 2019).

Much research on turmeric varieties in Ethiopia has

been done on agronomy and effect of postharvest
operation on physicochemical quality parameters.

However, there is no study on the chemical

composition of essential oil and pharmacological
activities of turmeric varieties grown in Ethiopia.

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Objectives
General objective:

To quantify bioactive constituents and evaluate antioxidant and anti

bacterial activities of turmeric varieties grown in Ethiopia.

Specific objectives:

1. To compare curcuminoid content, total polyphenol and extract

yields of different turmeric varieties.

2. To determine the chemical composition of turmeric essential oil

3. To evaluate DPPH free radical scavenging capacity and

antibacterial activities against E. coli, S. aureus and Salmonella
typhimurium, for turmeric cultivated in Ethiopia.
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Material and Methods

Fresh Powder
rhizomes

Curing
Solvent Hydrodistilla
extraction tion

Anti- Essential oil

Drying
Antioxidant bacterial characterizat
activity ion

Grinding Anti-
Antioxidant bacterial
activity
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Measured Data

TPC of the turmeric extracts was determined following the

Singleton et al. (1999) procedure adopted by Tanvir et al. (2017)
using Folin-Ciocalteu reagent.
 It was expressed as mg GAE/gram of extracts on a DW basis of

plant material after plotting calibration curve for different

concentrations (5–30 μ g/mL) of Gallic acid.

Total curcuminoids content was determined by UV-Visible

spectrophotometry at 425 nm.
 Calibration curve was produced from serial concentrations of (1.6,

3.2, 4.8, 6.4 and 8.0 μ g/mL) of curcumin as the standard

Free radical scavenging was determined using DPPH reducing assay

 Vitamin C (ascorbic acid) was used as a positive control.

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Antibacterial activity

 Turmeric extracts were dissolved in DMSO to get 100 mg/mL

concentration.

 Sterile filter paper discs that have 8 mm diameter were impregnated

with 50 μ l of diluted extract solution and then incubated for 15 min.

 The serial dilution was then surface spread onto nutrient agar plates
with 0.1 ml of bacterial culture standardized to 0.5 McFarland standards
(1.5×108 CFU ml-1) and incubated at 37 °C for 24h.

 The inhibition zones' diameter was measured, and the mean value of
three independent experiments was used in the statistical analysis.

 The control discs contained DMSO only, and the standard antibiotic
gentamycin (10 μ g/disc) was included in the assay as a positive control.

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Analysis of essential oil content

□ GC fitted with quadrupole MS, and DB-5MS fused silica capillary

column (30 m×0.25 mm [internal diameter film thickness 0.25 mm]).

□ Characterization of chemical components from EO of turmeric samples

were done based on
 their retention time; retention indices determined using a

homologous series of n-alkanes (C 8 –C18) under the same

temperature-programmed conditions, co-injection with standards.

 by comparing their retention indices and mass spectra with National

Institute of Standards and Technology (NIST) and Wiley, and then by
comparing the given mass spectral data with literature data.

□ The relative amount of each component of the essential oil and its
fractions were expressed as a percentage of the peak area relative to
the total peak area.

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Statistical analysis

The analysis of variance was completed using SAS

version 9.4 software (SAS, 2014).

Multiple means comparison was completed using

Duncan's method (Montgomery, 2020) and letter
groupings were generated at the 5% level of
significance.

Excel (in Microsoft Office, Microsoft, WA, USA)

was used to produce the figures.

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Result and Discussion

Table 1. Mean total curcuminoids and the yield of different extracts

obtained from the three varieties.

Variety % Curcuminoids % Methanol % Essential oils

extract
Bonga51/71 6.49b 13.62a 6.27a
HT3 /2002 5.12c 10.58b 3.92b
Dame 6.81a 13.42a 6.37a

Within each column, means sharing the same letter are not significantly different at the 5% level.

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…Result and Discussion

Table 2.Mean concentrations of TPC, scavenging capacity and IC50 of

turmeric varieties obtained from the four treatments.

Total phenolic content % DPPH scavenging IC50

Treatment
(mg GAE /g DW) capacity of 16 μ g/mL (µg/mL)
Bonga 51/71 96.52a 40.80c 44.32a
HT3/2002 83.21b 43.68b 32.25b
Dame 97.55a 46.58a 23.05c
Control - 48.83a 17.81d

Within each column, means sharing the same letter are not significantly different at the 5% level.
DW= dry weight basis; IC 50 = Half maximal (antioxidant activity) effective concentration.

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…Result and Discussion

90
80
70
%Scavenging actities

60
50
40 Bonga 51/71
30 HT3/2002
20 Dame
10
0
0 16 33 50 66 100 130
Concentration (µg/ml)

Figure 1. DPPH radical scavenging activity of three turmeric

varieties at different concentration (µg/mL) in methanol

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…Result and Discussion

Table 3. Inhibition effect of turmeric extracts, having antibacterial
activity against test bacterial.
Treatment
Test Pathogen Essential Methanol Gentamicin DMSO
oils extract
S.aureus +ve +ve +ve -ve
E.coli -ve -ve +ve -ve
S. typhimurium -ve -ve +ve -ve

+ve=inhibit bacterial growth; -ve=not inhibit bacteria growth.

 Among the tested bacterias, S. aureus (Gram-positive) was the only sensitive
organism to the plant extracts, which is in agreement with previous reports
(Norajit et al., 2007; Chouhan et al., 2017).
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…Result and Discussion

□ The varying degrees of sensitivity of the bacterial test organisms may be

due to the intrinsic tolerance of microorganisms (Naz et al., 2010).

□ The resistance of Gram-negative bacteria against turmeric extracts is due

to the rigid lipopolysaccharide on an outer membrane (Chouhan et al.,
2017).

□ The presence of lipoteichoic acids in Gram-positive bacteria, i.e., their

lipophilic ends in the (less complex, single-layer) membranes facilitates
the penetration of hydrophobic compounds present in essential oils.

□ The increased membrane permeability, consequently inducing leakage of

ions and important cell contents, finally leading to cell death (Stanojević et
al., 2015).
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…Result and Discussion

Z o n e o f In h ib itio n (m m )

Essential oil Methanol extract

a
9.00 b
8.00 c
7.00
6.00 d d d
5.00
4.00
3.00
2.00
1.00
0.00
Bonga 51/71 HT3/2002 Dame

Figure 2. Zone of inhibition (mm) by essential oil and methanol extract from different
turmeric varieties against S. aureus (Gram-positive bacteria).

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…Result and Discussion

Table 4. Constituents of essential oil obtained from Dame and Bonga51/71
Composition (%)
S/No Constituent identified a Molecular formula RI b Dame Bonga 51/71
1 α-Phellandrene C10H16 9.4 0.57 0.41
2 o-Cymene C10H14 10.4 0.36 0.24
3 Eucalyptol C10H18O 10.8 0.40 0.21
4 Trans- α-Bergamotene C15H24 25.06 0.12 0.13
5 Caryophyllene C15H24 25.61 0.36 0.39
6 (E)- β-Famesene C15H24 26.54 0.22 0.17
7 β-Curcumene C15H24 27.38 0.13 0.15
8 α-Curcumene C15H22 27.50 4.20 3.74
9 α-Zingiberene C15H24 28.0 4.96 4.53
10 β-Sesquiphellandrene C15H24 29.1 5.51 4.85
11 Caryophyllene oxide C15H24O 30.5 0.44 0.41
12 Humulene epoxide I C15H24O 30.7 0.17 0.19
a
The compoundsare listed in the order of elution from a DB-5MS column.
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b
Retention indices (RI) relative to C8-C18 n-alkanes on the DB-5MS column.
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…Result and Discussion

Table 4. Continue….
Composition (%)
S/No Constituent identified a Molecular formula RI b Dame Bonga 51/71
13 (E)-Nuciferol C15H22O 32.8 0.90 0.64
14 (Z)-α-trans-Bergamotene C15H24 33.0 0.99 0.62
15 β-Bisabolene C15H24 28.5 1.24 1.02
16 ar-Turmerol C15H22O 31.6 1.21 0.95
17 Bergamotol, Z- α -trans- C15H24O 32.98 0.99 0.62
18 7-epi-cis-sesquisabinene hydrate C15H26O 33.6 0.56 0.45
19 ar-Turmerone C15H20O 36.9 25.20 25.47
20 Tumerone C15H22O 37.3 32.41 35.16
21 Curlone C15H22O 39.8 17.98 18.19
22 6R, 7R-Bisabolone C15H24O 43.7 0.27 0.25
23 (Z)-α-Atlantone C15H22O 44.7 0.59 0.58
24 (E)-Atlantone C15H22O 47.1 0.67 0.76
a
The compoundsare listed in the order of elution from a DB-5MS column.
b
Retention indices (RI) relative to C8-C18 n-alkanes on the DB-5MS column. 19
 …Result and Discussion

O O O

Ar-Turmerone -Turmerone -Turmerone (= Curlone)

Figure 3: Chemical structures of key volatile components in the essential oil in

turmeric

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11/03/2021

Conclusions

 The investigation on turmeric varieties has shown that Dame variety

exhibited a prominent antioxidant and antibacterial activities followed
by Bonga 51/71variety.

 In vitro test on antibacterial activities of turmeric EOs and methanol

extract confirmed S. aureus was the only susceptible to extracts among
three bacterial strains.

 The other finding of this study revealed that, curlone, ar-turmerone and
tumerone were the major components of EOs.

 It is possible to say, this research will provide information on the

functional and therapeutic potential of turmeric from Ethiopia.
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Acknowledgements

TNSRC
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