Transcript Sukkari

Neuropharmacological &
analgesic properties of Ajwah,
Safawi and Sukkari
Omer A. A. Hamdi*, Bassem Y. Sheikh2 Jamil A Shilpi3
1 Head of Alneelain Centre
For natural products research and drug discovery,
1Department of Chemistry, Faculty of Science and Technology. Alneelain
University, Khartoum,Sudan.
2Department of Surgery, College of Medicine, Taibah University, Saudi Arabia
3 Pharmacy Discipline, Khulna University, Bangladesh
Acknowledgements
Al-Moalim MA Bin Ladin
(MABL) chair for Scientific
Miracles of Prophetic
Medicine, College of Medicine,
Taibah University
(Research grant no. MABL
37/02)
Facts: Date palm
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Fruits of Phoenix dactylifera L. (Arecaceae)
Origin: Middle East
Staple food in Middle East region
Due to its nutritional value, it was
naturalised in different parts of the world.
• Current status: More than 2000 cultivars
grow around the world
Narration of date palm in Al Quran and Al Hadith
• “And from the fruits of date-palms and grapes,
you derive strong drink and a goodly provision.
Verily, therein is indeed a sign for people who
have wisdom” (Surat An‐Nahl: 67).
• “He who eats seven dates of Madina (Ajwah
dates) every morning, will not be affected by
poison and magic on the day he eats them”
(Sahih Al-Bukhari).
Ethnobotanical uses
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Liver disorders
Diabetes
Constipation, diarrhoea
To reduce wrinkling of the skin
To relieve asthma
To alleviate headache
Expectorant
Ameliorating in cough, bronchitis, respiratory disorders
Increase immunity
Aphrodisiac, to treat sexual debility
Reported biological activity
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Antioxidant, antimutagenic
Antihaemolytic
Antiviral
Antifungal
Anti-inflammatory
Antihyperlipidemic
Hepatoprotective
Nephroprotective
Gastroprotective
Anticancer
Immunostimulating
Gonadotropic
Chemical constituents
• Simple phenolics
– p-hydroxy benzoic acid, protocatechuic acid, gallic acid, vanillic
acid, syringic acid
• Phenylpropanoids
– cinnamic acid, caffeic acid, o-caffeoyl shikimic acid, ferulic
acid, sinapic acid, o-coumaric acid, p-coumaric acid
• Carotenoids
– β-carotene, lutein
• Sterols
– cholesterol, campesterol, stigmasterol, β-sitosterol,
isofucosterol
• Flavonoids
– catechin, epi-catechin, quercetin, luteolin, apigenin
• Procyaninidins and anthocyanins
Background of present investigation
• In Ayurveda date palm is known as Kharjura and is
indicated for the treatment of
– psychosis, anxiety, cognitive dysfunction
– many of the nervous system disorders
• The fruit is also used alone or in combination to
treat
– sciatica, headache, hemicranias
– applied externally for inflammatory conditions
including abscess, boils and ulcers
Background of present investigation
• Literature survey on date palm revealed that
some Chinese and Japanese patented herbal
preparations containing date palm as one of the
component is indicated for
–treating sleeping disorders
Present investigation
• All these observations prompted us, as a
part of our research on Prophetic
medicine, to evaluate and compare
– neuropharmacological and
antinociceptive effects of
– Ajwah, Safawi and Sukkari
Ajwah
• 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.
Safawi
• Safawi is another popular date cultivar
growing in Almadinah Almunawarah, Saudi
Arabia. Safawy is oval shaped soft, moist
variety of dates with dark brown texture.
Sukkari
• Sukkari 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 chewy flesh. It grows mainly in
Qassim, Saudi Arabia.
Collection and extraction
• The dried ripe (in tamar stage) dates were
purchased from local date market in Al Madinah
Al Munawarah, KSA.
• The dates were identified by taxonomists at at
Taibah University.
• The dried dates were mashed with the help of
mortar and pestle, soaked in ethanol for 3 days
with periodic sonication.
• The extracts were filtered and dried using a
rotary vacuum evaporator at 45°C.
• The extracts were further freeze dried to get the
crude extract.
Test animal
• Young Swiss Albino mice
– 4-5 weeks old and weighing 20-25 g
• Purchased from the Animal Resources Branch
of ICCDR,B Bangladesh
• Acclimatisation with the laboratory condition
– Temperature 25±2°C
– Relative humidity: 56-60%
– 12 h dark-light cycle
Pentobarbitone-induced sleeping time test
• Test mice orally treated with date extracts
• After 30 min, pentobarbitone (50 mg/kg, i.p.)
was administered to induce sleep
• Time to onset for sleep recorded
• Duration of sleep recorded
• What to observe: CNS active compound will
• ↑ or ↓ time for onset of sleep
• ↑ or ↓ duration of sleep
Effect on pentobarbitone-induced sleeping time
Treatment
(n= 5)
Dose
(mg/kg)
R/A
Onset of sleep (min)
Duration of sleep
(min)
Control
10 ml/kg
p.o.
9.6±0.55
74.0±2.0
5
250
500
250
500
250
500
i.p.
p.o.
p.o.
p.o.
p.o.
p.o.
p.o.
3.6±0.34d
7.8±0.36cd
6.3±0.35ad
8.1±0.39c
7.0±0.35cd
8.6±0.34c
7.4±0.24cd
140±2.2f
90±2.4ce
110±2.2cf
86±2.3cd
100±3.0cf
83±1.6
97±2.6ce
Diazepam
Ajwah
Safawy
Sukkari
ap<0.05
vs D, bp<0.01 vs D, cp<0.001 vs D, dp<0.05 vs C, ep<0.01 vs C, fp<0.001 vs C; C=control, D=diazepam.
Results: The date extracts↓ time for onset of sleep and ↑ duration of sleep
i.e., showed CNS relaxing effect
Open field test: test for locomotor activity
• Test mice orally treated with date extracts
• Placed on a floor divided in squares
• Number of squares visited recorded for 3 min
(observation period 3 h)
• What to observe: CNS active compound will
• ↑ or ↓ number of squares visited
Results of open field test
Treatment
Dose
(n= 5)
(mg/kg)
0 min
30 min
60 min
90 min
120 min
180 min
Control
10 ml/kg
133±2
123±3
113±2
104±2
106±5
95±2
5
126±3
39±1f
30±1f
28±3f
29±1f
27±1d
250
128±2
89±2cf
74±3cf
73±2ce
75±3cf
81±3cd
500
129±2
84±1cf
71±2cf
67±1cf
71±1cf
74±2cf
250
139±2
91±3ce
78±3cf
76±4cf
79±3ce
84±4cd
500
133±3
85±1cf
73±2ce
70±2cf
75±2cf
80±4cf
250
129±4
93±2cf
82±3cf
75±3ce
73±3cf
82±2ce
500
137±3
86±3cf
77±3cf
72±3cf
69±3cf
79±2ce
Diazepam
Ajwah
Safawy
Sukkari
ap<0.05
Number of movement
vs D, bp<0.01 vs D, cp<0.001 vs D, dp<0.05 vs C, ep<0.01 vs C, fp<0.001 vs C; C=control, D=diazepam.
Inference: The date extracts reduced the locomotor activity in test mice
Hole board test: test for exploratory behaviour
• Test mice orally treated with date extracts
placed on a board having 16 evenly placed
holes
• Head dipping through the holes recorded for 2
min (observation period 3 h)
• What to observe: CNS active
compound will
• ↑ or ↓ number of head
dipping
Results of hole board test
Treatment
(n= 5)
Dose
(mg/kg)
0 min
30 min
Control
10 ml/kg
19±0.9
21±1.3
27±1.4
29±1.6
31±1.4
33±1.3
5
250
500
250
500
250
500
20±1.0
20±0.8
20±1.2
20±1.0
19±0.8
20±1.0
20±0.9
11±0.9f
17±1.1cd
16±1.2be
17±0.9cd
17±1.0ce
18±0.7cd
17±1.0cd
6±1.0f
14±0.8cf
12±0.9cf
18±0.7cf
15±0.9cf
18±0.8ce
15±0.9cf
6±0.8f
14±0.8cf
12±0.8cf
16±0.5cf
13±1.0cf
17±0.7cf
15±0.7cf
6±0.6f
17±0.7ce
14±1.0cf
19±0.9ce
16±1.1cf
20±0.8cf
17±1.1cf
7±0.5f
24±0.7ce
20±0.9cf
23±1.0cd
20±0.9cf
23±0.9cd
21±1.0ce
Diazepam
Ajwah
Safawy
Sukkari
ap<0.05
Number of head dipping
60 min
90 min
120 min
180 min
vs D, bp<0.01 vs D, cp<0.001 vs D, dp<0.05 vs C, ep<0.01 vs C, fp<0.001 vs C; C=control, D=diazepam.
Inference: The date extracts reduced the exploratory behaviour in test mice
Acetic acid induced writhing: test for analgesic
activity
• Intraperitoneal administration of 0.7% acetic
acid causes writhing (a sign of pain) in mice
• Number of writhing is counted
• What to observe: Analgesic compound will ↓
number of writhing
• Peripheral mechanism of
pain !
Effects on acetic acid induced writhing
Treatment
Dose
(n= 5)
(mg/kg)
Control
10 ml/kg
Diclofenac sodium
25
Ajwah
250
500
Safawy
250
500
Sukkari
250
500
ap<0.05
Number of writhing
33.0±1.0
9.4±0.5d
23.0±0.4cd
21.0±0.6cd
24.0±0.4cd
22.0±0.5cd
25.0±0.7cd
23.0±0.6cd
vs DS, bp<0.01 vs DS, cp<0.001 vs DS, dp<0.001 vs C; C=control, DS=diclofenac sodium.
Inference: The date extracts reduced peripherally mediated pain sensation in test
mice induced by acetic acid
Hot plate test: test for analgesic activity
• Mice placed on hot plate maintained at the
temperature of 55±0.5oC
• Paw licking or jumping is a sign of pain caused
by heat
• What to observe: Analgesic compound will ↑
response time
• Central mechanism of
pain !
Results of hot plate test
Treatment
(n= 5)
Control
Morphine
Ajwah
Safawy
Sukkari
ap<0.05
Dose
(mg/kg)
10 ml/kg
5
250
500
250
500
250
500
0 min
4.6±0.13
4.7±0.15
4.3±0.10
4.3±0.1
4.6±0.15
4.6±0.15
4.2±0.12
4.4±0.14
Response time (sec)
30 min
60 min
90 min
4.5±0.26
4.5±0.18
4.2±0.32
8.9±0.16f
11.4±0.40f 11.0±0.36f
5.7±0.24cf
5.9±0.14cf
5.0±0.10ce
5.9±0.27cf
7.0±0.19cf
6.6±0.20cf
5.7±0.17cf
6.0±0.13cf
5.3±0.19cf
6.6±0.20cf
7.3±0.14cf
6.5±0.21cf
5.2±0.12c
5.6±0.15cf
4.9±0.23c
5.9±0.17cf
6.9±0.15cf
5.8±0.12cf
120 min
4.4±0.15
8.7±0.20f
4.4±0.15c
5.2±0.10cd
4.3±0.14c
4.5±0.20c
4.3±0.17c
4.5±0.17cf
vs M, bp<0.01 vs M, cp<0.001 vs M, dp<0.05 vs C, ep<0.01 vs C, fp<0.001 vs C; C=control, M=morphine.
Inference: The date extracts reduced centrally mediated pain sensation in test mice
HPLC analysis for polyphenolic constituents
• Detection for major bioactive polyphenolic
constituents in date extracts by• DionexUltiMate 3000 Rapid Separation LC system
equipped with
– quaternary rapid separation pump (LPG-3400RS)
– acclaim® C18 column (4.6 × 250 mm; 5µm, Dionex USA)
– in a temperature-controlled column compartment (TCC3000) maintained at 30°C
– and photodiode array detector (DAD-3000RS)
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.
HPLC chromatogram of Ajwah extract
Peaks 1: (+)-catechin; 2: (-)-epicatechin; 3: trans-ferulic acid; 4: rosmarinic acid
HPLC chromatogram of Safawy extract
Peaks 1: (+)-catechin; 2: (–)-epicatechin; 3: trans-ferulic acid
HPLC chromatogram of Sukkari extract
Peaks 1: caffeic acid; 2: p-coumaric acid; 3: trans-ferulic acid
Contents of polyphenolic compounds
in date extracts
Polyphenolic
compound
Content in mg/100 g of dry extract*(% RSD)
Ajwah
Safawi
Sukkari
trans-Ferulic acid
11.70 (0.18)
5.01 (0.06)
2.28 (0.06)
(+)-Catechin
14.67 (0.29)
42.25 (0.57)
-
(–)-Epicatechin
9.15 (0.11)
21.93 (0.34)
-
Rosmarinic acid
3.73 (0.04)
-
-
Caffeic acid
-
-
3.11 (0.09)
p-Coumaric acid
-
-
1.37 (0.05)
*n=5; RSD: Relative standard deviation.
Conclusion
• Ajwah, Safawy and Sukkari dates have some degree of
relaxing effect on the brain.
• Reduced locomotor activity, and exploratory behaviour in
test mice suggest reduced CNS activity in treated mice.
• The extracts showed both centrally and peripherally acting
analgesic activity.
• In all cases, the effect was milder as compared to positive
control, indicating a moderate level of activity.
• Thus dates could be useful in producing mild relaxing effect
on the brain.
• The effects were similar with all the three date cultivars,
but relatively stronger with Ajwah dates.