ANTIOXIDANT ACTIVITIES OF FOREST HONEY

IN SOME REGIONS IN SUMBAWA DISTRICT

(Aktivitas Antioksidan Madu Hutan Di Beberapa Kecamatan Di Kabupaten Sumbawa)

By: Dinar Suksmayu Saputri and Yolli Eka Putri

Faculty of Agricultural Technology, Sumbawa University of Technology

Abstract
Honey is a natural substance produced by honey bees from flower nectar and plant
secretions. Honey contains minerals, proteins, vitamins, organic acids, flavonoids, phenolic
compounds, and enzymes such as catalase, peroxide, glucose oxidation, and other
phytochemicals, honey also has enzymatic and non-enzymatic antioxidant activity. The main
composition of honey consists of 80% carbohydrates, 17-20% water, 1-2% minerals and
organic compounds.
This study aims to identify the antioxidant content of forest honey in several regions
in Sumbawa Regency. Analysis of antioxidant activity, total phenol, extraction of phenolic
compounds, and identification of phenolic compounds in honey were done to identify the
antioxidant content.
Honey samples were obtained from forest sites in Lape, Tepal, Punik and Moyo
Island. The results of the study showed that forest honey in some of these areas contained
phenolic compounds (gallic acid) of 0,0633 – 0,3875 mg/g and antioxidant activity between
3,3365 – 30,9680% with IC50 values of 60,2 – 572,3 mg / mL .

Keywords: forest honey, antioxidants, Sumbawa

I. Background
Honey is a natural substance produced by honey bees from flower nectar and plant
secretions. Honey is known to have enzymatic and non-enzymatic antioxidant activity. In
addition, honey is a very saturated sugar solution, contains minerals, proteins, vitamins,
organic acids, flavonoids, phenolic compounds, and enzymes such as catalase, peroxide,
glucose oxidation, and other phytochemicals. Depending on geographical and climatic
conditions, various types of honey contain various phytochemicals including polyphenols and
phenolic acids which act as antioxidants. The main polyphenols in honey are flavonoids with
a content that varies between 60 and 460 µg per 100g of honey (Sime et al , 2007).
Research on honey has been widely done, but research and special studies of local
honey produced from Indonesian honey bees, especially Sumbawa Island, are still rarely
found. In addition, research and studies on local honey are important because the composition
of honey, including antioxidant content, is influenced by differences in plant species, climate,
and environmental conditions. Sumbawa's vast and lush island forest is an abundant food
source for honey-producing bees. Sumbawa forest honey is mostly produced by Apis
dorsata bees .
Taking honey in the forest is a side income for farmers on the island of
Sumbawa. Sumbawa forest honey has been believed to have good health benefits for a long
time. It is hoped that this research can prove the antioxidant activity of Sumbawa forest
honey. By knowing the antioxidant activity of Sumbawa honey in counteracting free radicals,
it is hoped that it will increase the economic value of Sumbawa honey and provide more
detailed information to consumers about the healthy effect of Sumbawa honey.

Translated from original article published in Jurnal Tambora Vol. 2 No. 3 December 2017.
(http://jurnal.uts.ac.id/index.php/Tambora/article/view/170)
II. Methods
Study design, study area and period
The study was conducted at the Laboratory of Food Chemistry Sumbawa University
of Technology, Laboratory of Food Analysis – Faculty of Agricultural Technology Jember
University and Laboratory of Food Quality and Food Safety Department of Agricultural
Product Technology Brawijaya University. The study starts in January 2017, ends
in November 2017.
Materials and tools
The materials used are forest honey from Lape, Batu Lanteh, Labuhan Badas and
Moyo Island regions. The chemicals used include Folin-Ciocalteau reagent, Na2 CO3, gallic
acid, DPPH reagent, 0,1 N HCl, (NH4 )2SO4, starch indicator and 0,1 N iodine. The tools used
in this study include test tubes, beakerglass, spectrophotometers, burettes, pipettes, analytical
balance, etc.
Antioxidant Activity Analysis (Chua, LS et al ., 2013)
0,75 mL of honey and methanol solution with a concentration of 20 - 40 mg/mL added with
1,5 mL of DPPH solution, then the absorbance was measured at a wavelength of 517 nm.
Analysis of Total Phenols (Marshall, SM, 2013)
5 g honey dissolved with 25 mL aquadest. Each sample was then added with the Folin-
Ciocalteau reagent and Na2CO3 (15%). The solution was incubated at room temperature for 30
minutes and the absorbance measured at 765nm wavelength.
Vitamin C Analysis ( Siti, N. et al., 2016)
10 g of honey is added to 50 mL measuring flask and added distilled water to the boundary
line. The sample is filtered to separate the filtrate. 5 mL filtrate is taken using a volume
pipette, put in an erlenmeyer, 2 drops of starch and 20 mL aquadest are added. The sample is
titrated with 0,1 N iodine solution until the color turns solid blue.

III. Results and Discussion

Antioxidant Activity
Honey antioxidant activity analysis was carried out using the DPPH (2,2-Diphenyl -1-
picrylhydrazyl) method. DPPH is a free radical that is stable at room temperature and
produces a purple solution in alcohol. DPPH will be reduced by the presence of
antioxidants. Analysis of antioxidant activity in honey samples is determined based on the
ability of antioxidants in honey to fight free radicals (DPPH). The electrons in the unpaired
DPPH will form bonds with hydrogen donated by antioxidants in honey. This reaction will
change the purple color of DPPH solution to yellow. The degree of color change is measured
using UV- visible spectrophotometer to determine the antioxidant activity of honey (Pontis,
JA et al ., 2014; Chua et al ., 2013).
The analysis found that the percent of free radical inhibition of Sumbawa honey was
3,34% for Tepal honey, 3,24% for Punik honey, 9,87% for Lape honey and 30,97% for Moyo
Island honey. Percent of free radical inhibition by Sumbawa honey can be seen in the graph
below,

Translated from original article published in Jurnal Tambora Vol. 2 No. 3 December 2017.
(http://jurnal.uts.ac.id/index.php/Tambora/article/view/170)
 35 30.97

% Free Radical Inhibition

30
25
20
15
9.87
10
3.34 3.24
5
0
Tepal Punik Lape Moyo Island
Sample Region

Figure 1. Antioxidant Activity of Sumbawa Honey

These results indicate that the antioxidant activity of honey samples taken from
several regions in Sumbawa is smaller than the honey in other studies. Honey from Apis
dorsata bees in Malaysia has 59,89% free radical inhibition; 21,06 – 32,14% in Java and
Sumatra honey; 17,7 – 82,68% in gelam and pineapple trees in Malaysia; 57,5% in Indian
honey and 44,55% in Algerian honey (Moniruzzaman et al. (2013); Chayati et al . (2014);
Husein et al. (2011)). Antioxidant activity of a material is influenced by the content of
phytochemical compounds that have antioxidant properties. These compounds include
phenolic acids, flavonoids, enzymes (glucose oxidase and catalase), ascorbic acid,
carotenoids, organic acids, amino acids and proteins. The antioxidant activity of the phenolic
component contributes to human health (Khalil, MI et al ., 2010).
Antioxidant activity is also expressed in the IC50 parameter. IC50 states the
concentration of the sample needed to inhibit 50% of free radicals, which in this case is
DPPH. The lower the IC50 value, the smaller the concentration of the sample used to ward
off 50% of free radicals, which means that the sample is stronger in warding off free radicals
(Pontis et al . 2014). IC50 values of honey samples from several regions in Sumbawa can be
seen in the graph below:

Translated from original article published in Jurnal Tambora Vol. 2 No. 3 December 2017.
(http://jurnal.uts.ac.id/index.php/Tambora/article/view/170)
 700

600 572.29 562.3

500

IC 50 400
304.93
300

200

100 60.2

0
Tepal Punik Lape Moyo island
Sample Region

Figure 2. IC50 Value of Sumbawa Honey

Total Phenol
Analysis of phenolic compounds were using the Follin-Ciocalteau method. The
reaction between the Follin-Ciocalteau reagent with phenolic compounds will form a
complex blue color that can absorb radiation (Pontis, JA et al. , 2014). The standard phenol
compound used in this analysis is gallic acid. From the analysis, it was found that total phenol
honey from Tepal was 0,0704 mg/g, 0,063 mg/g in Punik, 0,2927 mg/g in Lape and 0,3875
mg/g in Moyo Island.
0.45
0.39
0.40
0.35
Total Phenol (mg/g)

0.29
0.30
0.25
0.20
0.15
0.10 0.07 0.06
0.05
0.00
Tepal Punik Lape Pulau Moyo
Figure 3. Total Phenol (mg / g) of Sumbawa honey
Sample Region
The total value of honey phenols taken from several regions in Sumbawa is almost the
same as the results of other honey studies. Total of phenol Tualang honey were 0,22 – 0,38
mg/g; 0,17 – 1,32 mg/g in Japan multifloral honey; 0,23 – 0,73 mg/g in Portuguese honey;
0,29 mg/g in Echium vulgare; 0,03 – 0,16 mg/g in Czech honey and 0,25 – 0,54 mg/g in
Brazilian honey (Khalil et al. (2012); Kumazawa et al. (2012); Ferreira et al., (2009);
Nagai et al., (2012); Lachman et al., (2010); Pontis et al., (2014)).
Although total phenol content of Sumbawa honey were not much different from
honey from other regions, its antioxidant activity were distinct compared to other
studies. This is due to many disorders in the Follin - Ciocalteau method. Disturbances in this
method include the Follin - Ciocalteau reagent (a mixture
of phosphotungstic acid and phosphomolibdic ) which also reacts with compounds other than
phenol compounds such as vitamin C, so the total phenol yield becomes excessive. Other
reducing compounds such as sugar and amino acids can also interfere with analysis. This
causes a positive error ( positive error ) on the results (Ferreira et al ., 2009).

Translated from original article published in Jurnal Tambora Vol. 2 No. 3 December 2017.
(http://jurnal.uts.ac.id/index.php/Tambora/article/view/170)
 The content of phenol compounds and antioxidant activity in honey is directly
proportional to the color of honey. Honey color is related to the presence of pigments such as
carotenoids and flavonoids. Darker colored honey has higher total phenol and antioxidant
activity (Kumazawa et al ., 2012).
Vitamin C
Ascorbic acid (vitamin C) acts as an antioxidant by donating two electrons from the
double bond between the 2nd and 3rd carbon, donating these electrons which prevents other
compounds from oxidizing. This vitamin C oxidation produces new free radical compounds,
namely semidehydroascorbic or ascorbyl radicals which are more stable than other free
radicals and do not last long (Padayatty, SJ et al ., 2013; Siti, N. et al., 2016).
The content of vitamin C in honey from several districts in Sumbawa Regency can be
seen in the graph below:
35 32.72

30
Vitamin C (mg/100g)

25
20
15
10.13 10.33
10 7.18
5
0
Tepal Punik Lape Moyo island
Sample Region

Figure 6. Vitamin C Content (mg/100g) in Honey Samples

From the graph it can be seen that vitamin C content in honey samples were
consecutive from Moyo Island, followed by samples from Lape, Punik and Tepal
region. These results are in accordance with the antioxidant activity of honey, where the
antioxidant activity (% inhibition and IC50) of honey from Moyo Island is highest compared
to other regions of honey samples. But these results are smaller than previous
studies. Vitamin C in Tualang, Gelam and acacia honey from Malaysia contains 52,2 – 67,4
mg/100g). Honey from Nigeria contains 18,52 – 25,16 mg/100g of vitamin C. Vitamin C in
honey were varied by storage and processing, so it is rarely detected in honey that has
undergone processing such as screening and heating (Buba, F. et al . , 2013; Chua, LS et al .,
2013).

IV. CONCLUSION
Conclusions obtained from the study of the Antioxidant Activities Of Forest Honey
In Some Regions In Sumbawa Besar as follows:
a. Sumbawa forest honey from Moyo Island has higher phenol compounds and antioxidant
activity than honey from Lape, Tepal and Punik.
b. Antioxidant activity, the content of phenolic compounds and vitamin C of forest honey in
some regions in Sumbawa Besar Regency is smaller than other areas.
c. Need to improve the quality of Sumbawa forest honey

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V. REFERENCES
Campone, L. et al., 2014. Determination of Phenolic Compounds in Honey Using
Dispersive Liquid-Liquid Microextraction. Journal of Chromatography A 1334: 9 –
15
Chayati, I. dkk., 2014. Kandungan Komponen Fenolat, Kadar Fenolat Total dan
Aktivitas Antioksidan Madu dari Beberapa Daerah di Jawa dan Sumatera. MGMI
Vol. 6 No.1: 11 – 24.
Buba, F. et al., 2013. Analysis of Biochemical Composition of Honey Samples From
North-East Nigeria.Biochemistry and Analytical Biochemistry Vol. 2 Issue 3.
Chua, L.S., Rahaman, N.L.A., Adnan, N.A., and Tan, T.T.E., 2013. Antioxidant Activity
of Three Honey Samples in Relation with Their Biochemical Components. Journal
of Analytical Methods in Chemistry Vol. 13: 1-8
Darmawan, S. dan Agustarini, R., 2011. Penurunan Kadar Air Madu Hutan Alam
Sumbawa. Prosiding Seminar Nasional Peranan Hasil Litbang HHBK dalam
Mendukung Pembangunan Kehutanan: 313-321
Ferreira, I.C.F.R. et al., 2009. Antioxidant Activity of Portuguese Honey Samples:
Different Contributions of the Entire Honey and Phenolic Extract. Food Chemistry
Vol. 114 issue 4: 1438 -1443
Hussein, S.Z., Yusoff, K.M., Makpol, S., and Yusof, Y.A.M., 2011. Antioxidant
Capacities and Total Phenolic Contents Increase with Gamma Irradiation in Two
Types of Malaysian Honey. Molecules Vol. 16: 6378-6395
Khalil, M.I., Sulaiman, S.A., and Boukraa, L., 2010. Antioxidant Properties of Honey
and its Role in Preventing Health Disorder. The Open Nutraceuticals Journal Vol.
3: 6-16
Khalil, MD.I. et al., 2012. Content and Antioxidant Properties of Processed Tualang
Honey Collected From Different Regions in Malaysia. International Journal of
Pharmacy and Pharmaceutical Sciences Vol. 4: 214 – 219.
Kumazawa, S. et al., 2012. Antioxidant Activity in Honeys of Various Floral Origins:
Isolation and Identification of Antioxidants in Peppermint Honey. Food Science
and Technology Research Vol. 18 (5): 679 – 685.
Lachman, J. et al., Contents of Major Phenolic and Flavonoid Antioxidants in Selected
Czech Honey. Czech Journal of Food Science Vol. 28: 412 - 426
Marshall, S.M., 2013. Identification and Concentration of Phenolic and Carbonyl
Compounds in Florida Honeys. Thesis-University of Florida: Florida
Moniruzzaman, M. et al., 2013. Physicochemical and Antioxidant Properties of
Malaysian Honey Produced by Apis cerana, Apis dorsata and Apis melifera. BMC
Complementary and Alternative Medicine Vol. 13: 1 - 12
Nagai, T. et al., 2012. Functional Property of Honey from Echium vulgare. Food and
Nutrition Science Vol. 3: 614 – 620
Nascimento, D.L. et al., 2012. Extreme Effects of Season on The Foraging Activities and
Colony Productivity of a Stingless Bee (Melipona asilvai Moure, 1971) in
Northeast Brazil. Psyche: A Journal of Entomology Vol. 2012 artile ID 267361
Neupane, B.P. et al., 2015 Antioxidant Properties of Honey From Different Altitudes of
Nepal Himalayas. Polish Journal of Food and Nutrition Science Vol. 65 No.2: 87 -
91
Pontis, J.A., Costa, M.A., Silva, S.J.R., and Flach, A., 2014. Color, Phenolic, Flavonoid
Content and Antioxidant Activity of Honey from Roraima, Brazil. Food Science and
Technology Vol. 34(1): 69-73
Translated from original article published in Jurnal Tambora Vol. 2 No. 3 December 2017.
(http://jurnal.uts.ac.id/index.php/Tambora/article/view/170)
Prica, N. et al., 2014. Moisture and Acidity as Indicators of the Quality of Honey
Originating From Vojvodina Region. Arhiv Veterinarske Medicine Vol. 7 No.2: 99
-109
Puskadija, Z. et al., 2007. Influence of Weather Conditions on Honey Bee Visits (Apis
melifera carnica) During Sunflower (Helianthus annuus L.) Blooming Period.
ISSN 1330-7142
Sime, D., Atlabachew, M., Abshiro, M. R., and Zewde, T., 2015. Total Phenols and
Antioxidant Activities of Natural Honeys and Propolis Collected from Different
Geographical Regions of Ethiopia. Bull Chem Soc Ethiop. 29 (2): 163-172.
Siti, N. dkk., 2016. Penetapan Kadar Vitamin C pada Jerami Nangka (Artocarpus
hetertopophyllus L.). Jurnal Farmasi Sains dan Praktis Vol.II No1
Trimanto dan Sialiaan, F.A., 2017. Botanical Survey in Moyo Island, West Nusa
Tenggara, Indonesia: Inventory of Flora Collection at Forest. The Journal of
Tropical Life Science Vol. 7 No.2: 158 – 166
Wiriadinata, H. dkk. 2013. Floristic Study of West Sumbawa, Indonesia. A Journal of
Taxonomic Botany, Plant Sociology and Ecology Vol. 13(5): 391-455.

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(http://jurnal.uts.ac.id/index.php/Tambora/article/view/170)