ISSN: 2347-467X, Vol. 08, No. (1) 2020, Pg.

155-163

Current Research in Nutrition and Food Science

www.foodandnutritionjournal.org

The Effect of Cultivar Variation on Total Phenolic

Contents and Antioxidant Activities of Date
Palm Fruit (Phoenix dactylifera L.)
JEERAWAN HINKAEW, YURAPORN SAHASAKUL,
NATTAPOL TANGSUPHOOM and UTHAIWAN SUTTISANSANEE*

Institute of Nutrition, Mahidol University, Phutthamonthon, Nakhon Pathom, Thailand.

Abstract
Date palm fruit (Phoenix dactylifera L.) is generally consumed in form
of dry fruits (Tamr stage); however, fresh date palm fruit in Khalal stage
is currently of interest due to its high total phenolic contents (TPCs) Article History
and antioxidant activities. Therefore, this study aimed to investigate
the water based extraction conditions such as temperature (30-90°C), Received: 29 October 2019
Accepted: 20 March 2020
shaking time (0.5-6 hours) and solid-to-liquid ratio (100-500 mg/mL)
of date palm fruit in Khalal stage. This will help to establish extraction Keywords
protocol for future food applications. Under optimized extraction
conditions, TPCs and antioxidant activities of cell culture originated (CO) Antioxidant Activities;
Cultivar Variation;
and seed originated (SO) date palm fruit were compared to examine Extraction Conditions;
the effect of cultivar variation. The results suggest that optimization of Khalal Stage;
extraction conditions was reached using the concentration of 100 mg/ Phoenix dactylifera L.
mL, extraction temperature of 50°C, and shaking time of one hour. Under
these extraction conditions, CO exhibited TPCs of 3.47±0.33 mg GAE/g
DW as well as antioxidant activities of 0.0025±0.00, 16.13±0.81 and
123.21±9.77 µmol TE/g DW measured by DPPH radical scavenging,
FRAP and ORAC assays, respectively. Interestingly, SO exhibited
higher TPCs (3.87±0.23 mg GAE/g DW) and antioxidant activities
(0.0021±0.00, 19.23±0.80 and 185.68±9.29 µmol TE/g DW by DPPH
radical scavenging, FRAP and ORAC assays, respectively).The findings
from this study will promote consumption of date palm fruits in Khalal
stage as healthy food and support its future development for food.
Also, this will enhance the growth of date palm fruit with its attendant
economic benefits.

CONTACT Uthaiwan Suttisansanee uthaiwan.sut@mahidol.ac.th Institute of Nutrition, Mahidol University, Phutthamonthon,

Nakhon Pathom, Thailand.

© 2020 The Author(s). Published by Enviro Research Publishers.

This is an Open Access article licensed under a Creative Commons license: Attribution 4.0 International (CC-BY).
Doi: http://dx.doi.org/10.12944/CRNFSJ.8.1.14
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 156

Introduction temperatures, shaking time and solid-to-liquid ratio

Date palm (Phoenix dactylifera L.) is a largely of date palm fruit at Khalal stage in Barhi cultivar
agricultural plant in the Middle East and North Africa (collected in Thailand in 2019) to establish the proper
with more than 1,500 varieties.1 The development of extraction protocols regarding bioactive compounds
date palm fruit can be divided into 5 stages, including and antioxidant activities. Under optimized extraction
Hanabeuk, Kimri, Khalal, Rutab and Tamr stages.2 conditions, TPCs and antioxidant activities of date
During fruit development, the color changes from palm fruit from different cultivar originates, like cell
green to yellow or red, depending on the varieties.2 culture originated (CO) and seed originated (SO),
In general, the last three stages namely Khalal, Rutab were compared to examine the effect of cultivar
and Tamr stages are commonly consumed because variation.
of softer texture and sweeter flavor. The fruits in Tamr
stage are consumed in a form of dry fruits; however, Materials and Methods
fresh date palm fruit in Khalal stage is currently of Sample Preparation
interest. The previous study stated that Khalal stage Cell culture originated (CO) and seed originated
of Barhi cultivar, which is recently economical crop (SO) date palm fruits at Khalal stage in Barhi cultivar
growing in Eastern Thailand, is currently consumed were supplied by T.A.P. Chon Buri Co., Phanat
in the form of fresh fruit.2 Besides, date palm fruit in Nikhom district, Chon Buri province, Thailand. SO
this stage is a good source of nutritive values referred was received from date palms growing from seeds of
from the proximate composition study, in which it cell cultured date palms. The samples were collected
contained 222 kcal energy, 1.8 g protein, 0.2 g fat, during July-August, 2019. The samples were
56.9 g carbohydrate, and 39.7% moisture content per cleaned with deionized water (DI) before separating
100 g.1 Interestingly, previous study also suggested flesh and seed. The colors of fresh samples were
that the fruit in Khalal stage exhibited higher total analyzed using a ColorFlex EZ spectrophotometer
phenolic content and antioxidant activity analyzed from Hunter Associates Laboratory (Virginia, USA)
by FRAP assay than the ones in Rutab and Tamr and expressed in the Hunter-Lab units, including
stages.3-4 In term of health properties, date palm L* (darkness to lightness), a* (green to red) and
fruit was reported to contain high phenolic contents b* (blue to yellow). Clean samples were cut into
with antioxidant activities, which can fight against 0.3 cm thick before freeze-drying using a Heto
various toxicants.5 Moreover, in vitro study stated powerdry PL9000 freeze dryer (Heto Lab Equipment,
that date palm fruit extract inhibited human breast Allerod, Denmark) for 3 days. Dr y samples
adenocarcinoma progression and exhibited the were ground into fine powder using a grinder
properties against leukemia and microbial activities.6 (Philips 600W series from Philips Electronic Co.,
Ltd., Jakarta, Indonesia). The powdery samples were
Current studies from Thailand on the effect of Khalal packed into vacuum aluminum foil bag and kept at
stage of date palm cultivar variation on TPCs and -20°C until further analysis.
antioxidant activities is sparse. Normally, date palm is
populated using cell culture technique to sustainably Optimization of Extraction
maintain the genetics of exact cultivar. However, Optimization of extraction conditions were performed
this technique required high preparation cost and is under three parameters, including shaking times,
time consuming. Thus, reproducing date palm using extraction temperatures and concentrations
seed can reduce cost and is a custom propagation of sample, according to the report by Sripum
to increase date palm population. However, seed et al., 2016.7 Briefly, CO powder was mixed with
originated date palm might possess variation in distilled water before incubating in a temperature-
genetics due to natural selection, possibly in attempt controlled water bath shaker (WNE45 series from
to self-adapt to the environment. Besides, proper Memmert GmBh, Wisconsin, USA) for a particular
extraction conditions for further development of food shaking time period (0.5, 1, 2, 4 and 6 hours).
product from the fruit at this stage is missing. The mixture was then centrifuged at 3,800xg using
a Hettich® ROTINA 38R refrigerated centrifuge
Therefore, this study aimed to investigate the water (Andreas Hettich GmbH, Tuttlingen, Germany) for
based extraction conditions, such as extraction 15 minutes. The supernatant was filtered through a
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 157

0.45 µm PES membrane syringe filter before keeping 95% (v/v) aqueous ethanol (200 µL). The mixture
at -20°C for further analyses. was incubated at room temperature for 30 minutes
in dark. The reaction was monitored at a wavelength
To investigate the shaking time, the sample at the of 520 nm using the 96-well microplate reader. The
concentration of 500 mg/mL were extracted at 30°C free radical scavenging activity was calculated using
and shaking for 0.5, 1, 2, 4 and 6 hours. Likewise, the following equation;
extraction temperatures were optimized by varying
extraction temperatures at 30, 50, 70, 90°C, while antioxidant activity = (∆A-I /S ) x F / DW x fv / 1000,
fixing the sample concentration at 500 mg/mL and
shaking for 1 hour. Lastly, the optimized sample where ∆A is a different absorbance between blank
concentrations were examined by varying the sample and sample, I is an intercept of standard curve, S is
concentrations to 100, 200, 300, 400 and 500 mg/ a slope of standard curve, F is a dilution factor, fv is
mL, while fixing the extraction temperature at 50°C a final volume of solvent extract (mL), DW is a dry
and shaking for 1 hour. The optimized extraction weight of sample (g DW) and 1000 is a conversion
conditions were determined through TPCs and from L to mL. Trolox (10-640 µM) was used as a
FRAP activity as indicated below. standard.

Total Phenolic Contents (TPCs) Assay Oxygen Radical Antioxidant Capacity (ORAC)
TPCs were determined according to Ainsworth and Assay
Gillespie, 20078 and Sripum et al., 2016.7 The extract The ORAC assay was determined according to
(25 μL) was mixed with 10% (v/v) Folin–Ciocalteu’s Huang et al., 200211 and Sripum et al., 2016.7
phenol reagent (50 μL) and 7.5% (w/v) sodium In brief, the extract (25 µL) was mixed with 30 nM
carbonate (200 μL). The mixture was then incubated fluorescein solution (150 µL) before incubating at
at 25°C for 2 hours in dark. TPCs were monitored 37°C for 30 minutes in dark. To the mixture, 153 mM
at a wavelength of 765 nm using a SynergyTM HT 2,2'–azobis (2–amidinopropane) dihydrochloride
96-well UV-visible microplate reader from BioTek (25 µL) was added, and the fluorescence intensity
Instruments, Inc. (Vermont, USA) with Gen 5 data was measured for 90 minutes using the microplate
analysis software. Gallic acid (0-200 μg/mL) was reader at the excitation wavelength of 485 nm and
used as a standard. the emission wavelength of 528 nm. The antioxidant
activity was calculated by the difference in area
Ferric Reducing Antioxidant Power (FRAP) under sodium fluorescein decay curve (AUC) using
Assay the following equation;
FRAP assay was performed according to Benzie
and Strain, 19969 and Sripum et al., 2016.7 The AUC = (0.5 + f1/f0 + f2/f0 + f3/f0 + …. + (0.5)fi/f0) x CT,
extract (20 μL) was incubated at 25°C for 8 minutes
in dark with FRAP reagent (150 μL), which was where f 0 is the initial fluorescence reading at
prior prepared by mixing 300 mM acetate buffer pH 0 minute, f i is the fluorescence reading at i
3.6, 10 mM TPTZ solution in 40 mM HCl, 20 mM minutes, and CT is the cycle time in minutes. Trolox
ferric chloride hexahydrate (FeCl3•6H2O) solution in (0-100 µM) was used as a standard.
the ratio of 10:1:1, respectively. The FRAP activity
was monitored at a wavelength of 600 nm using the Results and Discussion
microplate reader. Trolox (0–250 µM) was used as Characteristics of Date Palm Fruit
a standard. It was found that the overall size of SO was smaller
than CO (Fig. 1). The average size of CO was
1,1–Diphenyl–2–Picryl Hydrazyl (DPPH) Radical 3.73±0.12 cm in height, 2.11±0.09 cm in width and
Scavenging Assay 0.67±0.24 cm in thickness, while SO possessed
DPPH radical scavenging assay was performed 3.13±0.28 cm in height, 2.00±0.90 cm in width
according to Fukumoto and Mazza, 2000 10 and 0.54±0.10 cm in thickness. The results of color
and Sripum et al., 2016. 7 The sample extract measurement expressing as Hunter-Lab units
(22 µL) was mixed with 135 µM DPPH reagent in (L*, a* and b*) suggested that both CO and SO
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 158

possessed the dark reddish yellow color of exocarb similar color, but the exocarp of SO was yellower
(outer skin), while mesocarb (inner flesh) had lighter than that of CO.
color (Table 1). The mesocarp of CO and SO had

Fig. 1: Cell culture originated (left) and seed originated (right) date palm fruits at
Khalal stage in Barhi cultivar. The line indicated the scale of 1 centimeter

Table 1: Color values of cell culture originated (CO) and seed originated
(SO) date palm fruits at Khalal stage in Barhi cultivar

Date palm fruit Color values in Hunter-Lab units

L* a* b*

CO Exocarp 48.10±1.39 12.21±0.57 37.28±0.08

Mesocarp 65.72±2.99 3.21±0.79 20.85±1.93
SO Exocarp 48.26±4.89 11.80±0.92 46.61±2.65
Mesocarp 65.77±1.35 3.87±0.78 21.02±2.05

All data were expressed as mean ± standard deviation (SD) of triplicate experiments.

Fig. 2: Effects of various shaking times (0.5-6.0 hours) on TPCs and antioxidant activities through
FRAP assay under fixed extraction temperature at 30°C and extract concentration at 500 mg/mL.
The capital and small letters indicated significant difference at p < 0.05 of TPCs and FRAP
values, respectively, using one-way ANOVA with Duncan’s multiple comparison tests
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 159

Effects of Extraction Time were decreased. It was previously suggested by

The result indicated that by fixing the extract Fick’s second law of diffusion that a final equilibrium
concentration at 500 mg/mL and extraction can be achieved at a certain time when the solute
temperature at 30°C, the shaking time at 1 hour concentrations in the solid matrix and in the bulk
exhibited the highest TPCs (1.35 mg GAE/g solution are equal.13 Thus, after the equilibrium is
DW) and FRAP activity (6.98 µmol TE/g DW) reached, increase in time cannot increase released
(Fig. 2). Similarly to the previous study of Lapornik phenolics. Instead, prolonged incubating time might
et al., 200512 in the comparison of extraction time cause a degradation of phenolics.14 Thus, in our
and bioactive compound of red currant (Ribes experiment, an equilibrium time was reached at
rubrum var. Rondom), it was found that 1 hour 1 hour, while longer shaking time from 2-6 hours
for extraction provided the highest content of caused a reduction in TPCs and FRAP activities
phenolics and antioxidant activity. After increasing (Fig. 2).
the shaking time, phenolics and antioxidant activity

Fig. 3: Effects of various extraction temperatures (30-90°C) on TPCs and antioxidant activities
through FRAP assay under the fixed shaking time at 1 hour and extract concentration at 500 mg/
mL. The capital and small letters indicated significant difference at p < 0.05 of TPCs and FRAP
values, respectively, using one-way ANOVA with Duncan’s multiple comparison test

Effect of Extraction Temperature FRAP assay.16 The temperature affects bioactive

Under the fixed shaking time at 1 hour and extract compounds by softening the tissue of plant
concentration at 500 mg/mL, no significant difference cell wall and stimulates the phenol-protein and
in TPCs of the samples extracted under 30-70°C phenol-polysaccharide to diffuse into the solvent.17
was observed (approx. 1.5 mg GAE/g DW), while However, heating might induce phenolic compound
TPC was decreased significantly at 90°C (Fig. 3). degradation as well.17 Supported by the study of Le et
Antioxidant activity determined by FRAP assay al., 2005 stating that after increasing the temperature
was potentially the highest when extracting at 50°C from 19 to 65°C for extraction of citrus peels, FRAP
(5.17 µmol TE/g DW) (Fig. 3). Therefore, the most activities were reduced by half.15 Therefore, the
suitable extraction temperature for date palm increased extraction temperature with prolonged
fruit was 50°C. Corresponding with this study, period of time can affect the activity of antioxidant
the previous studies reported that extraction activity.
temperature of Henna (Lawsonia inermis) at
55°C provided the greatest TPCs,15 and peach Effect of Sample Concentrations
(Prunus persica L.) extracted at 50-60°C was By fixing shaking time at 1 hour and extraction
found to exhibit the highest antioxidant activity via temperature at 50°C, date palm fruit extracted
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 160

at 100 m g /m L p rov i d e d th e h i g h e s t TP C black mulberr y leaves extracted under the

(1.8 mg GAE/g DW), while increased extract concentration of 0.033 mg/mL exhibited the higher
concentrations (200-500 mg/mL) continuingly TPCs than the one extracted at 0.1 mg/mL. The
lowered TPCs (Fig. 4). Likewise, date palm fruit solid-to-liquid ratio was previously reported to
extracted at 100 and 200 mg/mL provided the be consistent with the mass transfer principle.19
highest FRAP activities (approx. 12 µmol TE/g DW). The driving force during mass transfer is consistent
However, increasing extract concentrations from 300 with the concentration between the solid and
to 500 mg/mL caused a decrease in FRAP activities. liquid solvent.13-14,20 Low solid-to-liquid ratio can
According to the previous study on the effect of increase surface interactions between solid and
solid-to-liquid ratio (12.5-100 mg/mL) of citrus peel, solvent, resulting in more driving force for bioactive
it was found that the lowest extract concentration at compounds to penetrate through cell walls into
12.5 mg/mL provided the highest TPCs.18 Likewise, solvent.

Fig. 4: Effects of various concentrations (100-500 mg/mL) on TPCs and antioxidant activities
through FRAP assay under the fixed shaking time at 1 hour and extraction temperature at 50°C.
The capital and small letters indicated significant difference at p < 0.05 of TPCs and
FRAP values, respectively, using one-way ANOVA with Duncan’s multiple comparison tests

Table 2: Total phenolic contents (TPCs) and antioxidant activities of date palm fruit in
Barhi cultivar with cell culture originated (CO) and seed originated (SO) methods

Originates
TPCs Antioxidant activities
(mg GAE/g DW)
FRAP assay DPPH radical scavenging ORAC assay
(µmol TE/g DW) assay (µmol TE/100 g DW) (µmol TE/g DW)

CO 3.47±0.33b 16.13±0.81b 0.25±0.02a 123.21±9.77b

SO 3.87±0.23a 19.23±0.80a 0.21±0.01b 185.68±9.29a

The small letters indicated significantly difference between cultivar methods using independent
t-test at p < 0.05.

TPCs and FRAP activities of CO and SO samples 50°C of extraction temperature), the difference in
under the optimized extraction conditions TPCs (determined by Folin Ciocalteau’s reagent)
(100 mg/mL extract, 1 hour of shaking time and and antioxidant activities through FRAP, ORAC and
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 161

DPPH scavenging activity assays of CO and SO date palm fruits at Khalal stage but in different
were investigated. The results suggested that SO cultivars (Khalas and Sequh cultivars with the TPCs
exhibited significantly higher TPCs and antioxidant of 0.03 and 0.05 mg GAE/g DW, respectively).26
activities determined by FRAP and ORAC assays Date palm fruit at Khalal stage in Tunisian and
than CO (Table 2). However, CO exhibited higher Ahmar cultivars, however, exhibited the similar TPCs
antioxidant activity determined by DPPH radical (3.0-8.5 mg GAE/g DW) as the ones in our
scavenging assay than SO. experiments.27 Nevertheless, the different TPCs
might be a result of altered extraction conditions
Two functions of antioxidants, hydrogen atom and plant varieties.
transfer (HAT) and single electron transfer (SET)
mechanisms are proposed.21-22 The HAT reaction Conclusion
is based on a transfer of hydrogen atom (H•) of Date palm fruit at Khalal stage in Barhi cultivar is
antioxidant to free radical, while SET reaction generally consumed as fresh fruit. The optimized
involves the donation of one electron to electron extraction conditions were achieved at 100 mg/
acceptor. In our experiments, TPCs were correlated mL extract concentration, 1 hour of shaking time
with FRAP (SET mechanism) and ORAC (HAT and 50°C of extraction temperature. Under these
mechanism) assays, suggesting that phenolics from conditions, seed originated date palm fruits exhibited
date palm fruits can function as both SET and HAT potentially higher TPCs and antioxidant activities
antioxidants. However, DPPH radical scavenging than cell culture originated samples. This study has
(HAT and SET mechanisms) activity was in the demonstrated the nutritional benefit of this fruit due
opposite trend. It is possible that this method is to its antioxidant properties. It is equally important
less sensitive than FRAP and ORAC assays; to encourage the growth of the plant due to its
therefore, small change in DPPH radical scavenging enormous economic benefit.
activity can lead to misinterpreted results. Besides,
DPPH radical scavenging assay is more suitable Acknowledgments
for measuring antioxidant activity in hydrophobic The authors would like to express gratitude to T.A.P.
system.23 The in vitro investigation on Chaetoceros Chon Buri Co., Phanat Nikhom district, Chon Buri
didymus extract suggested that increased TPCs province, Thailand, for providing the date palm
could be achieved with increased polarity index of fruit. We also would like to thank Miss Varittha
solvent extraction, while the highest DPPH radical Sritalahareuthai and Mr. Werawat Wannasaksri for
scavenging activity was observed with the sample insight suggestions of manuscript preparation.
extracted under non-polar solvent.24
Funding
Comparing to previous literature using the sample at The author(s) received no financial support for the
the same stage (Khalal stage) and cultivar (Barhi), research, authorship, and/or publication of this
it was found that TPCs detected in our experiment article.
were higher than those reported in the ethanolic and
acetone extracts (2.15 and 3.16 µmol GAE/g fresh Conflict of Interest
weight, respectively).25 Moreover, the TPCs detected The authors declare no conflict of interest.
in our experiment were higher than the ones from

References

1. Al-Farsi A. M., Lee Y. C. Nutritional and 3. Haider S. M., Khan A. I., Jaskani J. M., Naqvi
Functional Properties of Dates: a Review. A. S., Khan M. M. Biochemical Attributes
Crit Rev Food Sci Nutr. 2008; 48(10): 877-887. of Dates at Three Maturation Stages.
2. Ghnimi S., Umer S., Karim A., Kamal-Edin Emir J Food Agr. 2014; 26(11): 953-962.
A. Date Fruit (Phoenix dactylifera L.): an 4. Amira A. E., Guido F., Behija E. S., Manel
Underutilized Food Seeking Industrial I., Nesrine Z., Ali F., et al. Chemical and
Valorization. Nutr Food Sci. 2017; 6: 1-10. Aroma Volatile Compositions of Date Palm
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 162

(Phoenix dactylifera L.) Fruits at Three 2007; 55(3): 381-387.

Maturation Stages. Food Chem. 2011; 127(4): 14. Salar R. K., Purewal S. S., Bhatti M. S.,
1744-1754. Optimization of Extraction Conditions and
5. EI-Far H. A., Oyinloye E. B., Sepehrimanesh Enhancement of Phenolic Content and
M., Allah G. A. M., Abu-Reidh I., Shaheen Antioxidant Activity of Pearl Millet Fermented
M. H., et al., Novel Findings and Future with Aspergillus Awamori MTCC-548. Reffit.
Directions for Food and Drug Discovery. Curr 2016; 2(3): 148-157.
Drug Discov Technol. 2019; 16(1):2-10. 15. Tan M. C., Tan C. P., Ho C. W. Effects
6. Jaouni A. K. S., Alghamdi N., Hossary E. D., of Extraction Solvent System, Time and
Harakeh S. Effects of Phoenix dactylifera Temperature on Total Phenolic Content of
Ajwa on Infection, Hospitalization, and Henna (Lawsonia inermis) Stems. Int Food
Survival Among Pediatric Cancer Patients Res. 2013; 20(6): 3117-3123.
in a University Hospital: A Nonrandomized 16. Mokrani A., Madani K. Effect of Solvent,
Controlled Trial. Integr Cancer Ther. Time and Temperature on The Extraction
2019; 18(1): 1-9 of Phenolic Compounds and Antioxidant
7. Sripum C., Kukreja R. K., Charoenkiatkul Capacity of Peach (Prunus persica L.) Fruit.
S., Kriengsinyos W., Suttisansanee U. The Sep Purif Technol. 2016; 162(1): 68-76.
Effect of Storage Conditions on Antioxidant 17. Bey M. B., Louaileche H., Zemouri S.
Activities and Total Phenolic Contents Optimization of Phenolic Compound
of Parboiled Ger minated Brown Rice Recovery and Antioxidant Activity of Light
(Khao Dok Mali 105). Int Food Res J. and Dark Dried Fig (Ficus carica L.) Varieties.
2016; 23(4): 1827-1831. Food Sci Biotechnol. 2013; 22(6): 1613-1619.
8. Ainsworth E. A., Gillespie, K. M. Estimation of 18. Xu G., Ye X., Chen J., Liu D. Effect of Heat
Total Phenolic Content and Other Oxidation Treatment on the Phenolic Compounds and
Substrates in Plant Tissues using Folin– Antioxidant Capacity of Citrus Peel Extract.
Ciocalteu reagent. Nat. 2007; 2(4): 875-877. J Agric Food Chem. 2007; 55(2): 330-335.
9. Benzie F. F. I., Strin J. J. I. The Ferric Reducing 19. RadojkoviĆ M., ZekoviĆ Z., JokiĆ S., VidoviĆ
Ability of Plasma (FRAP) as a Measure of S., LepojeviĆ Ž., MiloševiĆ S. Optimization
Antioxidant Power: the FRAP Assay. Anal of Solid-Liquid Extraction of Antioxidants
Biochem.1996; 239(1): 70-76. from Black Mulberry Leaves by Response
10. Fukumoto L. R., Mazza G. Assessing Surface Methodology. Food Technol.
Antioxidant and Prooxidant Activities of 2012; 50(2): 167-176.
Phenolic Compounds. J Agric Food Chem. 20. Cacace E. J., Mazza G. Mass Transfer
2000; 48(8): 3597-3604. Process during Extraction of Phenolic
11. Huang D., Ou B., Hampsch-Woodill M., Compounds From Milled Berries. J Food Eng.
Flanagan, A. J., Prior L. R. High-Throughput 2003; 59(4): 379-389.
Assay of Oxygen Radical Absorbance 21. Apak R., Özyürek M., Güçlü K., Çapanoğlu E.
Capacity (ORAC) Using a Multichannel Liquid Antioxidant Activity/Capacity Measurement.
Handling System Coupled with a Microplate 2. Hydrogen Atom Transfer (HAT)-Based,
Fluorescence Reader in 96-Well. J Agric Food Mixed-Mode (Electron Transfer (ET)/HAT),
Chem. 2002; 50(16): 4437-4444. and Lipid Peroxidation Assays. J Agric Food
12. Lapornik B. M., Prošek M., Wondra G. A. Chem. 2016; 64(5): 1028-1045.
Comparison of Extracts Prepared From Plant 22. Choi H., Yang K., Lee S. G., Lee J. P.,
By-Products Using Different Solvents and Koo I. S. Single Electron Transfer (SET)
Extraction Time. J Food Eng. 2005; 71(2): Pathway: Nucleophilic Substitution Reaction
214-222. of 4-Chloro-7-Nitrobenzofurazan with Anilines
13. Silva M. E., Rogez H., Larondelle Y. in MeOH-MeCN Mixtures. B Korean Chem
Optimization of Extraction of Phenolics Soc. 2010; 31(10): 2801-2805.
from Inga Edulis Leaves Using Response 23. Floegel A., Kim D. O., Chung S. J., Koo S.
Surface Methodology. Sep Purif Technol. I., Chun O. K. Comparison of ABTS/DPPH
 HINKAEW et al., Curr. Res. Nutr Food Sci Jour., Vol. 8(1), 155-163 (2020) 163

Assays to Measure Antioxidant Capacity in Food Chem. 2013; 61(18): 4278-4286.

Popular Antioxidant-Rich US Foods. J Food 26. Ali H., Al-Khalifa A. R., Farouk A., Shaheen M.
Compos Anal. 2011; 24(7): 1043-1048. Effect of Maturation Stages on Flavor Profile
24. Noreen H., Semmar N., Farman M., McCullagh and Antioxidant Activity of Date Palm Fruits
J. S. Measurement of Total Phenolic Content (Phoenix dactylifera) Grown in Saudi Arabia.
and Antioxidant Activity of Aerial Parts of Int J Pharmaco, 2018; 14(3): 407-414.
Medicinal Plant Coronopus didymus. Asian 27. Lemine F. M. M., Ahmed M. V. O. M.,
Pac J Trop Biomed. 2017; 10(8): 792-801. Maoulainine L. B. M., Bouna Z. E. A. O., Samb
25. Borochov-Neori H., Judeinstein S., Greenberg A., Boukhary A. O. M. S. O. Antioxidant Activity
A., Volkova N., Rosenblat M., Aviram, M. of Various Mauritanian Date Palm (Phoenix
Date (Phoenix dactylifera L.) Fruit Soluble dactylifera L.) Fruits at Two Edible Ripening
Phenolics Composition and Anti-Atherogenic Stages. Food Sci Nutr. 2014; 2(6): 700-705.
Properties in Nine Israeli Varieties. J Agric