تفاصيل البحث
Comparative study of neuropharmacological, analgesic properties and phenolic profile of Ajwah, Safawy and Sukkari cultivars of date palm (Phoenix dactylifera)
ورقة منشورة
7/27/2016 12:00:00 AM
كرسي المعلم محمد بن لادن لأبحاث الإعجاز العلمي في الطب النبوي
Received: 13 December 2015 /Accepted: 27 July 2016 /Published online: 16 August 2016
# The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract In addition to the rich nutritional value, date palm is
also used in various ethnobotanical practices for the treatment
of various disease conditions. Present investigation was un-
dertaken to examine the neuropharmacological and
antinociceptive effect of the ethanol extract of three date cul-
tivars growing in Saudi Arabia, namely Ajwah, Safawy and
Sukkari. Neuropharmacological effect was observed by pen-
tobarbitone induced sleeping time, open field, and hole board
test. Antinociceptive activity was tested by acetic acid induced
writhing and hot plate test. The date extracts were also sub-
jected to HPLC analysis to detect the presence of common
bioactive polyphenols. All the three date extracts extended
the pentobarbitone induced sleeping time, reduced locomotor
activity in open field test and reduced exploratory behaviour
in hole board test in mice. The extracts also reduced acetic
acid induced writhing and delayed response time in hot plate
test. The activities were stronger for Ajwah than the other two
date cultivars. HPLC analysis indicated the presence of trans-
ferulic acid in all three cultivars, while (+)-catechin and (−)-
epicatechin only in Ajwah and Safawy. The observed neuro-
pharmacological and analgesic activity could be partly due to
the presence of (+)-catechin, (−)-epicatechin and trans-ferulic
acid, three important plant polyphenols well known for their
neuroprotective activity and their ability to exert antioxidant
activity on brain cells. Present investigation also supports the
ethnobotanical use of date palm to provide ameliorating ef-
fects in pain and CNS disorders.
Keywords Date palm . Open field test . Hole board test .
(+)-catechin . (−)-epicatechin . Trans-ferulic acid
Introduction
The ripe fruits of Phoenix dactylifera L. (Arecaceae), also
known as date palm, plays an important role in social and
economic perspective of the people living in the oasis of the
Middle East by the virtue of its nutritional and pharmacolog-
ical properties (Baliga et al. 2011). The fruit serves as an
important source of nutrition in an arid region hostile to hab-
itation of plants. It is believed that the date palm originated in
the Middle East. Due to its rich food value, date was later
naturalised in many parts of the world, and at present more
than 2000 cultivars of P. dactylifera are known to grow around
the globe (Guido et al. 2011). Apart from its use as a staple
food, date palm enjoys its use in the ethnomedicinal practice
for a wide range of ailments. Date palm is used for the treat-
ment of liver disorders (Gill 1992), diabetes (Ziyyat et al.
1997), constipation, diarrhoea (Hmamouchi 1999), and as an
aphrodisiac (Zaid and Arias-Jiménez 2002). Date fruits are
taken alone or in combination with other ingredients to get
relief from asthma (Zaid and Arias-Jiménez 2002), to reduce
wrinkling of the skin (Bauza et al. 2001), as an expectorant
Electronic supplementary material The online version of this article
(doi:10.1007/s13596-016-0239-5) contains supplementary material,
which is available to authorized users.
* Bassem Yousef Sheikh
consultprofsheikh@gmail.com
1 College of Medicine, Taibah University, PO Box 456, Almadinah
Almunawarah 41411, Saudi Arabia
2 Pharmacy Discipline, Life Science School, Khulna University,
Khulna 9208, Bangladesh
3 Department of Chemistry, Faculty of Science and Technology,
Alneelain University, 11121 Khartoum, Sudan
4 BCSIR Laboratories, Bangladesh Council of Scientific and Industrial
Research (BCSIR), Dhaka 1205, Bangladesh
Orient Pharm Exp Med (2016) 16:175–183
DOI 10.1007/s13596-016-0239-5
Online ISSN 2211-1069
Print ISSN 1598-2386
and ameliorating in cough, bronchitis, respiratory disorders, to
alleviate headache, to treat sexual debility and to increase
immunity (Selvam 2008; Zaid and Arias-Jiménez 2002).
Investigations revealed that date palm possesses antioxidant,
antimutagenic (Vayalil 2002), antihaemolytic (Abuharfeil
et al. 1999), antiviral (Jassim and Naji 2010), antifungal
(Shraideh et al. 1998), anti-inflammatory (Mohamed and Al-
Okabi 2004), antihyperlipidemic (Al-Maiman 2005), hepato-
protective (Al-Qarawi et al. 2004; Sheikh et al. 2014),
nephroprotective (Al-Qarawi et al. 2008), gastroprotective
(Al-Qarawi et al. 2005), anticancer (Ishurd and Kennedy
2005), immunostimulating (Puri et al. 2000), and gonadotrop-
ic (El-Mougy et al. 1991) activity. The date fruit is also rich in
pharmacologically important phytochemical constituents in-
cluding simple pheolics (p-hydroxy benzoic acid,
protocatechuic acid, gallic acid, vanillic acid, syringic acid),
phenylpropanoids (cinnamic acid, caffeic acid, o-caffeoyl shi-
kimic acid, ferulic acid, sinapic acid, o-coumaric acid, p-
coumaric acid) (Mansouri et al. 2005), carotenoids (β-caro-
tene, lutein), sterols (cholesterol, campesterol, stigmasterol, β-
sitosterol, isofucosterol) (Kikuchi and Miki 1978), flavonoids
and their glycosides (catechin, epi-catechin, quercetin,
luteolin, apigenin) (Hong et al. 2006), procyaninidins (Hong
et al. 2006), and anthocyanins (Al-Farsi et al. 2005).
The Sukkari date is the best-selling date in Saudi Arabia.
These golden-brown dates have patches of lighter colour and
are medium or small cone shaped with a firm exterior. This
date is characteristically sweet as compared to other cultivars
with its chewy flesh. It grows mainly in Qassim, Saudi Arabia.
Safawy is another popular date cultivar growing in Almadinah
Almunawarah, Saudi Arabia. Safawy is oval shaped soft,
moist variety of dates with dark brown texture. Unlike other
dates, Ajwah dates are relatively smaller in size. Ajwah is
round shaped, soft, dark brown coloured date which looks
almost black with fine texture and white wrinkles. Ajwah
has special interest to Muslims as it has been mentioned in
the Prophetic medicine.
In Ayurveda date palm is known as Kharjura and is indi-
cated for the treatment of psychosis, anxiety, cognitive dys-
function and many of the nervous system disorders
(Shanmugapriya and Patwardhan 2012). The fruit is also used
alone or in combination to treat sciatica, headache,
hemicranias, and applied externally for inflammatory condi-
tions including abscess, boils and ulcers (Shanmugapriya and
Patwardhan 2012). Literature survey on date palm revealed
that some Chinese and Japanese patented herbal preparations
containing date palm as one of the component can be benefi-
cial in treating sleeping disorders ( Katsumichi et al. 1997;
Tian 2014). Furthermore, acute toxicity study with date palm
extract prior to our project on biological investigation of date
extracts revealed extended period of sleep in test animals. All
these observations prompted us, as a part of our research on
Prophetic medicine (El-Ameen et al. 2015; Halabi and Sheikh
2014; Maulidiani et al. 2015; Taha et al. 2015), to evaluate and
compare neuropharmacological effects of two date cultivars
growing in Madinah (Ajwah and Safawy) and one growing
outside Madinah but within Saudi Arabia (Sukkari).
Materials and methods
Plant material and extraction
The dried ripe (in tamar stage) dates were purchased from
local date market in Al Madinah AlMunawarah, Saudi
Arabia. The dates were identified by taxonomists at
Bangladesh National Herbarium where a voucher specimen
(DACB 41158) has been submitted for future reference. For
easy identification by the readers, images have been given in
Fig. 1. The dried dates were mashed with the help of mortar
and pestle, soaked in ethanol for 3 days with periodic sonica-
tion. The extracts were filtered and dried using a rotary vacu-
um evaporator at 45 °C under reduced pressure to get semi-
solid masses. The extracts were further freeze dried to get the
crude extract.
Test animals
Young Swiss Albino mice of 4–5 weeks old and weighing 20–
25 g were purchased from the Animal Resources Branch of
International Centre for Diarrhoeal Disease Research,
Bangladesh (ICCDR,B). They were acclimatised with the lab-
oratory condition (temperature: 25 ± 2 °C, relative humidity:
56–60 %, 12 h dark-light cycle) before the commencement of
the pharmacological experiments.
Chemicals and drugs
Arbutin, benzoic acid, caffeic acid, (+)-catechin, trans-
cinnamic acid, p-coumaric acid, ellagic acid, (−)-epicatechin,
trans-ferulic acid, gallic acid, hydroquinone, kaempferol,
myricetin, quercetin, rosmarinic acid, rutin, syringic acid,
vanillic acid, and vanillin were purchased from Sigma-
Aldrich (St. Louis, MO, USA). Reference drugs were gener-
ously provided by Beximco Pharmaceuticals Ltd. Bangladesh
(diclofenac sodium and caffeine), Popular Pharmaceuticals
Fig. 1 Pictures of date palms. a: Ajwah; b: Safawy; c: Sukkari
176 B. Y. Sheikh et al.
Ltd. Bangladesh (morphine) and Incepta Pharmaceuticals Ltd.
Bangladesh (pentobarbitone).
Acute toxicity test
Test mice divided into different groups containing 6 mice of
either sex were treated with graded doses (62.5–4000 mg/kg
body weight) of date palm extract while the control group
received control vehicle (1 % Tween 80 in water) orally. The
animals were observed for 72 h and mortality, general signs
and symptoms of toxicity were recorded for each group
(Lorke 1983).
Grouping and dosing
Animals of either sex were randomly divided into four groups,
each comprising of six animals. Control group received vehi-
cle (1 % Tween 80 in water) orally at a volume of 10 ml/kg.
Test groups were pre-treated orally with three date extracts at
the doses of 250 and 500 mg/kg, while positive control group
received reference drug.
Pentobarbitone-induced sleeping time test
Test groups were orally treated with the three date extracts at
the aforementioned doses while control and positive control
group received control vehicle and diazepam (5 mg/kg, p.o.),
respectively. After thirty minutes, pentobarbitone (50 mg/kg,
i.p.) was administered to each mouse to induce sleep. The
latent period for the onset of sleep, and the duration of sleep
was recorded (Shilpi et al. 2004).
Open field test
Test mice and control mice were placed on the floor of an open
field of the dimension 100 cm × 100 cm × 40 cm, divided in
squares coloured black and white. The number of squares
visited by each group was recorded for 3 min after every thirty
minutes starting from the time of extract administration and
continued for a period of 4 h (Shilpi et al. 2004).
Hole board test
Each mouse from the control, test and positive control group
was placed in the hole board having 16 evenly placed holes.
Head dipping of the mouse through the holes was recorded for
2 min on every 30 min for a period of 2 h starting from the
time of extract administration (Mondal et al. 2014).
Acetic acid induced writhing test
Acetic acid (0.7 %, 10 ml/kg) was administered intraperitone-
ally to each mouse 30 min after the administration of control
vehicle, date extracts and diclofenac sodium (25 mg/kg, p.o.).
After 5 min of acetic acid administration, number of writhing
by each group was recorded for 10 min (Mondal et al. 2014).
Hot-plate test
Control vehicle, date extracts and morphine (5 mg/kg, i.p.)
treated mice were placed on a hot plate maintained at the
temperature of 55 ± 0.5 °C on every 30 min starting from
the time of extract administration and continued for a period
of 2 h to record response time, i.e., the time required for paw
licking or jumping. To avoid any injury or accidental paw
damage of the mouse, a cut-off point of 15 s was maintained
(Mondal et al. 2014).
HPLC analysis for polyphenolic constituents
Detection of the major polyphenolic compounds present in
the date extracts was conducted by HPLC analysis on a
DionexUltiMate 3000 Rapid Separation LC system
(Thermo Fisher Scientific Inc., MA, USA) equipped with
a quaternary rapid separation pump (LPG-3400RS), ac-
claim® C18 column (4.6 × 250 mm; 5 μm, Dionex USA)
housed in a temperature-controlled column compartment
(TCC-3000) maintained at 30 °C, and photodiode array
detector (DAD-3000RS) (Chuanphongpanich and
Phanichphant 2006; Islam et al. 2014). Separation was done
using a gradient elution programme consisting of
5%A95%B 0–9 min, 10%A80%B10%C 10–19 min,
20%A/60%B/20%C 20–30 min, followed by flushing and
further equilibriation with 100%A for 5 min; where A, B
and C are acetonitrile, acetic acid solution of pH 3 and
methanol, respectively. For detection, photodiode array de-
tector was set to the range of 200–700 nm for the entire
Table 1 Effects of three date extracts on pentobarbitone induced
sleeping time in mice
Treatment
(n = 5)
Dose
(mg/kg)
Route of
administration
Onset of
sleep (min)
Duration of
sleep (min)
Control (1 %
Tween 80 in
water)
10 ml/kg p.o. 9.6 ± 0.55 74.0 ± 2.0
Diazepam 5 i.p. 3.6 ± 0.34d 140 ± 2.2f
Ajwah 250 p.o. 7.8 ± 0.36cd 90 ± 2.4ce
500 p.o. 6.3 ± 0.35ad 110 ± 2.2cf
Safawy 250 p.o. 8.1 ± 0.39c 86 ± 2.3cd
500 p.o. 7.0 ± 0.35cd 100 ± 3.0cf
Sukkari 250 p.o. 8.6 ± 0.34c 83 ± 1.6
500 p.o. 7.4 ± 0.24cd 97 ± 2.6ce
a
p < 0.05 vs. diazepam, b p < 0.01 vs. diazepam, c
p < 0.001 vs. diazepam, d p < 0.05 vs. control, e
p < 0.01 vs.control, f
p < 0.001 vs. control
Comparison of neuro and analgesic activity of three date cultivars 177
experimental period while UV detector was set to 280 nm for 0–
18 min, 320 nm for 19–24 min, and 380 nm for 25–30 min.
Calibration curve was prepared using a standard solution of
methanol containing arbutin (5 μg/ml), benzoic acid
(8 μg/ml), caffeic acid (3 μg/ml), (+)-catechin (10 μg/ml), trans-
cinnamic acid (1 μg/ml), p-coumaric acid (2 μg/ml), ellagic acid
(10 μg/ml), (−)-epicatechin (5 μg/ml), trans-ferulic acid
(3 μg/ml), gallic acid (4 μg/ml), hydroquinone (4 μg/ml),
kaempferol (2 μg/ml), myricetin (4 μg/ml), quercetin
(2 μg/ml), rosmarinic acid (4 μg/ml), rutin (6 μg/ml), syringic
acid (3 μg/ml), vanillic acid (4 μg/ml), and vanillin (3 μg/ml).
Test solutions for date extracts were prepared at a concentration
of 5 mg/ml in methanol. The injection volume was 20 μl for
standard or extract solutions, and the experiments were conduct-
ed with a flow rate of 1 ml/min.
Statistical analysis
Results were expressed as mean ± SEM. One-way or two-
way ANOVA followed by Bonferroni’s test was done for
statistical analysis and results were considered significant
when p < 0.05.
Results
Results of acute toxicity test
No mortality or signs or symptoms of toxicities were observed
for any of the date extracts under investigation even at the
highest dose (4.0 g/kg) tested. However, at higher doses, the
mice showed extended sleeping tendency which persisted up
to 48 h.
Results of pentobarbitone-induced sleeping time test
All the date extracts reduced the time for the onset of sleep and
increased the duration of sleep as compared to the control with
the extent of intensity relatively higher with Ajwah than
Safawy or Sukkari, but less than that of diazepam. All the
results were statistically significant (Table 1).
Results of open field test
In the open field test, the extracts showed a decrease in the
movements in test mice as compared to control. The decrease
Table 2 Effects of three date extracts on open field test in mice
Treatment (n = 5) Dose (mg/kg) Number of movement
0 min 30 min 60 min 90 min 120 min 180 min
Control 10 ml/kg 133.2 ± 2.6 123.4 ± 3.23 113.0 ± 2.2 104.2 ± 2.3 106.8 ± 4.7 95.8 ± 2.0
Diazepam 5 126.2 ± 3.7 39.6 ± 1.7f 30.2 ± 1.0f 28.2 ± 2.5f 29.4 ± 1.2f 27.6 ± 1.2d
Ajwah 250 128.0 ± 2.0 89.4 ± 2.2cf 74.6 ± 2.7cf 73.2 ± 1.6ce 75.4 ± 2.8cf 81.0 ± 3.0cd
500 129.2 ± 2.2 84.4 ± 1.8cf 71.6 ± 1.9cf 67.0 ± 1.4cf 71.0 ± 1.0cf 74.2 ± 1.7cf
Safawy 250 139.4 ± 2.6 91.6 ± 3.9ce 78.0 ± 3.1cf 76.0 ± 3.8cf 79.0 ± 2.5ce 84.4 ± 3.7cd
500 133.0 ± 3.0 85.4 ± 1.9cf 73.4 ± 1.8ce 70.2 ± 1.9cf 75.2 ± 2.4cf 80.2 ± 3.8cf
Sukkari 250 129.4 ± 4.5 93.2 ± 2.8cf 82.2 ± 2.8cf 75.4 ± 2.7ce 73.0 ± 2.8cf 82.2 ± 1.9ce
500 137.2 ± 3.6 86.2 ± 3.7cf 77.0 ± 3.3cf 72.4 ± 3.3cf 69.4 ± 2.7cf 79.2 ± 2.2ce
a
p < 0.05 vs. diazepam, b p < 0.01 vs. diazepam, c
p < 0.001 vs. diazepam, d p < 0.05 vs. control, e
p < 0.01 vs. control, f
p < 0.001 vs. control
Table 3 Effects of three date extracts on hole board test in mice
Treatment (n = 5) Dose (mg/kg) Number of head dipping
0 min 30 min 60 min 90 min 120 min 180 min
Control 10 ml/kg 19.2 ± 0.9 21.4 ± 1.3 27.0 ± 1.4 29.4 ± 1.6 31.0 ± 1.4 33.4 ± 1.3
Diazepam 5 20.2 ± 1.0 11.4 ± 0.9f 6.0 ± 1.0f 6.4 ± 0.8f 6.2 ± 0.6f 7.4 ± 0.5f
Ajwah 250 20.4 ± 0.8 16.8 ± 1.1cd 14.6 ± 0.8cf 13.8 ± 0.8cf 17.0 ± 0.7ce 23.8 ± 0.7ce
500 20.4 ± 1.2 16.4 ± 1.2be 12.8 ± 0.9cf 12.4 ± 0.8cf 14.2 ± 1.0cf 19.8 ± 0.9cf
Safawy 250 19.8 ± 1.0 17.4 ± 0.9cd 18.0 ± 0.7cf 16.0 ± 0.5cf 19.4 ± 0.9ce 23.4 ± 1.0cd
500 19.2 ± 0.8 17.0 ± 1.0ce 15.0 ± 0.9cf 13.4 ± 1.0cf 16.2 ± 1.1cf 20.4 ± 0.9cf
Sukkari 250 20.4 ± 1.0 18 ± 0.7cd 18.4 ± 0.8ce 17.2 ± 0.7cf 20.4 ± 0.8cf 23.2 ± 0.9cd
500 20.2 ± 0.9 17.2 ± 1.0cd 15.4 ± 0.9cf 14.6 ± 0.7cf 16.6 ± 1.1cf 21.0 ± 1.0ce
a
p < 0.05 vs. diazepam, b p < 0.01 vs. diazepam, c
p < 0.001 vs. diazepam, d p < 0.05 vs. control, e
p < 0.01 vs. control, f
p < 0.001 vs. control
178 B. Y. Sheikh et al.
in the movement was prominent from 30 min until 120 min,
with the effect gradually fading at 180 min. Diazepam, used as
positive control showed similar results but the effect was
stronger as compared to the three dates extracts. All the results
were statistically significant (Table 2).
Results of hole board test
In the hole board test, a decrease in the number of head dip-
ping was observed for the test mice. Although, the effect was
not as strong as that of diazepam, the results of the date ex-
tracts were significantly different when compared to control
and the effect of Ajwah extract was stronger than the other two
date extracts (Table 3).
Results of acetic acid induced writhing
All three date extracts significantly reduced acetic acid in-
duced writhing in test mice as compared to the control.
Diclofenac sodium, used as the positive control in this study
showed strong analgesic activity (Table 4).
Results of hot plate test
The response time in test mice was extended by all the three
date extracts and morphine as compared to the control and the
results were statistically significant. Maximum effect was ob-
served one hour after the treatment, which gradually faded at
the end of the experiment (2 h) (Table 5).
Results of HPLC analysis
Results of HPLC analysis of the standards and three date
cultivars under investigation are presented in Figs. 2, 3, 4
and 5 and Table 6. All the three date cultivars showed the
presence of trans-ferulic acid with its highest content in
Ajwah. Among other phenolic components, (+)-catechin and
(−)-epicatechin were present in Ajwah and Safawy but not in
Sukkari. In contrast, caffeic acid and p-coumaric acid were
present only in Sukkari. Rosmarinic acid was only present in
Ajwah.
Discussion
The fruits of date palm have a long history of its use in tradi-
tional medicine. While date palm is reported to be used in
headache, recent study suggests that the fruits have
cerebroprotective activity in mice suffering from cerebral is-
chemia (Kalantaripour et al. 2012). It was also found to exhibit
neuroprotective activity in mice with ischemia induced bilat-
eral common carotid artery occlusion (Pujari et al. 2011).
Presence of flavonoids, sterols and ascorbic acid was credited
for the observed ameliorating effect. Present investigation was
done to evaluate neuropharmacological and antinociceptive
effects of three cultivars of date palm, namely Ajwah,
Safawy, and Sukkari. Extended period of sleeping by the test
mice in acute toxicity test suggests that the effect was not a
‘post lunch dip’ which might occur with high sugar content of
Table 4 Effects of three date extracts on acetic acid induced writhing in
mice
Treatment
(n = 5)
Dose (mg/kg) Number of writhing
Control (1 % Tween 80 in water) 10 ml/kg 33.0 ± 1.0
Diclofenac sodium 25 9.4 ± 0.5d
Ajwah 250 23.0 ± 0.4cd
500 21.0 ± 0.6cd
Safawy 250 24.0 ± 0.4cd
500 22.0 ± 0.5cd
Sukkari 250 25.0 ± 0.7cd
500 23.0 ± 0.6cd
a p < 0.05 vs. diclofenac sodium, b p < 0.01 vs. diclofenac sodium, c
p < 0.001 vs. diclofenac sodium, d p < 0.001 vs. control
Table 5 Effects of three date
extracts on hot plate test in mice Treatment (n = 5) Dose
(mg/kg)
Response time (sec)
0 min 30 min 60 min 90 min 120 min
Control 10 ml/kg 4.6 ± 0.13 4.5 ± 0.26 4.5 ± 0.18 4.2 ± 0.32 4.4 ± 0.15
Morphine 5 4.7 ± 0.15 8.9 ± 0.16f 11.4 ± 0.40f 11.0 ± 0.36f 8.7 ± 0.20f
Ajwah 250 4.3 ± 0.10 5.7 ± 0.24cf 5.9 ± 0.14cf 5.0 ± 0.10ce 4.4 ± 0.15c
500 4.3 ± 0.1 5.9 ± 0.27cf 7.0 ± 0.19cf 6.6 ± 0.20cf 5.2 ± 0.10cd
Safawy 250 4.6 ± 0.15 5.7 ± 0.17cf 6.0 ± 0.13cf 5.3 ± 0.19cf 4.3 ± 0.14c
500 4.6 ± 0.15 6.6 ± 0.20cf 7.3 ± 0.14cf 6.5 ± 0.21cf 4.5 ± 0.20c
Sukkari 250 4.2 ± 0.12 5.2 ± 0.12c 5.6 ± 0.15cf 4.9 ± 0.23c 4.3 ± 0.17c
500 4.4 ± 0.14 5.9 ± 0.17cf 6.9 ± 0.15cf 5.8 ± 0.12cf 4.5 ± 0.17cf
a
p < 0.05 vs. morphine, b p < 0.01 vs. morphine, c
p < 0.001 vs. morphine, d p < 0.05 vs. control, e
p < 0.01 vs.
control, f
p < 0.001 vs. control
Comparison of neuro and analgesic activity of three date cultivars 179
date palm extracts. All these extracts showed an increase in the
pentobarbitone induced sleeping time in mice. Pentobarbitone
is a barbiturate type sedative and hypnotic agent, which acts
through allosteric modification of GABA receptor resulting in
postsynaptic inhibition (ffrench-Mullen et al. 1993).
Neuroactive agents, depending on their stimulating or de-
pressing effect, can increase or decrease the duration of pen-
tobarbitone induced sleep in test animal. In our present study,
all the date extracts decreased the latency for the onset of
sleep, as well as increased the duration of sleep indicating that
the extracts might have some sedative effect on CNS. Open
field and hole board tests are important but simple ways of
determining CNS effect of any agent (Takagi et al. 1971;
Uddin et al. 2006). Results of the present investigation shows
a decrease in locomotor activity in test mice treated with date
extracts suggesting that the date extracts might have decreased
CNS activity in test mice. In both open field and hole board
test, the effect was highest with Ajwah extract. Present inves-
tigation suggests a relaxing effect in the test mice treated with
date extracts. It is well established that antioxidants play an
important role in reducing oxidative stress in brain and pro-
vide neuroprotective effect (Giacalone et al. 2011;
Mohamadin et al. 2010; Sheikh and Mohamadin 2012;
Wang et al. 2006). In our present investigation, two important
Fig. 2 HPLC chromatogram of a
standard mixture of polyphenolic
compounds. Peaks 1: arbutin; 2:
gallic acid; 3: hydroquinone; 4:
(+)-catechin; 5: vanillic acid; 6:
caffeic acid; 7: syringic acid; 8:
(−)-epicatechin; 9: vanillin; 10: p-
coumaric acid; 11: trans-ferulic
acid; 12: rutin; 13: ellagic acid;
14: benzoic acid; 15: rosmarinic
acid; 16: myricetin; 17: quercetin;
18: trans-cinnamic acid; 19:
kaempferol
Fig. 3 HPLC chromatogram of
Ajwah date extract. Peaks 1: (+)-
catechin; 2: (−)-epicatechin; 3:
trans-ferulic acid; 4: rosmarinic
acid
180 B. Y. Sheikh et al.
flavanols, namely, (+)-catechin, and (−)-epicatechin were de-
tected in Ajwah and Safawy extracts. Neuroprotective effect
of these two flavonols is well established and the mechanism
of action is believed to be their antioxidant activity and bene-
ficial actions on brain cells which include positive effects on
mood (Mandel and Youdim 2004; Nehlig 2013). Although
neuroprotection by hydroxycinnamic acids can be much less
as compared to catechins, the effect of trans-ferulic acid can-
not be ruled out since it is reported to exert neuroprotective
effect in in-vivo and in-vitro tests and its antioxidant capacity
might be the contributing factor for such activity (Cheng et al.
2008; Luo and Sun 2011; Wu et al. 2014). The traditional use
of date plam in headache prompted us to test the extracts for
antinociceptive activity. Decrease in the writhing in acetic acid
induced writhing test suggests that date extracts can show
analgesia through peripheral mechanism of pain inhibition,
i.e., block inflammatory pathway of pain sensation through
the inhibition of prostaglandin synthesis (Murata et al.
1997). This is in agreement with previous finding in which
methanol extract of Zaghlool dates showed anti-inflammatory
activity in rat model (Mohamed and Al-Okabi 2004). An in-
crease in the response time in hot plate test further suggests
that the observed analgesia might also involve centrally acting
mechanism (Wigdor and Wilcox 1987). In different studies,
Fig. 4 HPLC chromatogram of
safawy date extract. Peaks 1: (+)-
catechin; 2: (−)-epicatechin; 3:
trans-ferulic acid
Fig. 5 HPLC chromatogram of
Sukkari date extract. Peaks 1:
caffeic acid; 2: p-coumaric acid;
3: trans-ferulic acid
Comparison of neuro and analgesic activity of three date cultivars 181
trans-ferulic acid has showed analgesic activity in thermal
hyperalgesia, acetic acid induced writhing and mechanical
allodynia tests in mice (Lv et al. 2013; Ozaki 1992). Thus, trans-
ferulic acid could be credited to some extent for the observed
analgesic activity of the date extracts. In addition, more potent
activity of Ajwah compared to the two other date cultivars might
be due to the higher content of trans-ferulic acid in Ajwah. (+)-
Catechin, and (−)-epicatechin, detected in Ajwah and Safawy
extracts are also reported to show anti-inflammatory activity in
various in-vivo and in-vitro models including inhibition of NO
production and LPS-induced prostaglandin E2 release (García
et al. 2013; Wang and Cao 2014; Yang et al. 2015).
Conclusion
Present investigation suggests that Ajwah, Safawy and
Sukkari cultivars of date palm have some degree of relaxing
effect on the brain. It is possible that these extracts reduce
CNS activity resulting in decreased locomotor activity in test
mice. The extracts also produced analgesic activity in test
mice supporting its use in headache in traditional medicine.
However, in all cases, the effect was not as strong as that of the
positive control, indicating a moderate level of neuropharma-
cological and analgesic activity, and thus could be of interest
for producing mild relaxing effect on the brain. The effects
were similar with all the three date cultivars, but relatively
stronger with Ajwah dates.
Acknowledgments We are grateful to Al-Moalim MA BinLaden chair
for Scientific Miracles of Prophetic Medicine (MABL) for providing
financial support (research grant no. MABL 37/02) and Pharmacy
Discipline, Khulna University, Bangladesh for laboratory facilities to
conduct bioactivity studies.
Compliance with ethical standards
Ethical statement The experimental protocols were approved by the
Ethical Committee of Pharmacy Discipline, Life Science School, Khulna
University, Bangladesh.
Conflict of Interest The authors declare no conflict of interests.
Open Access This article is distributed under the terms of the Creative
Commons Attribution 4.0 International License (http://creativecommons.org/
licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the
original author(s) and the source, provide a link to the Creative Commons
license, and indicate if changes were made.
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