ISSN 2394-7330

International Journal of Novel Research in Healthcare and Nursing

Vol. 2, Issue 3, pp: (166-172), Month: September-December 2015, Available at: www.noveltyjournals.com

Anti-Inflammatory and Analgesic Activities of

Aqueous Extract Date Palm
(Phoenix Dactylifera L) Fruit in Rats
Umar Ibrahim Maryam 1, Nordin Simbak2, Abdulkareem Umar3, Ibrahim Haruna
Sani4, Atif Amin Baig5, Thant Zin6, Swethadri GKM7
1,3,4
Post graduate student, Faculty of Medicine, University Sultan Zainal Abidin, Terengganu Malaysia
2,5,6,7
Medical lecturers, Faculty of Medicine, University Sultan Zainal Abidin, Terengganu Malaysia

Abstract: Objective: The purpose of this study is to evaluate the anti-inflammatory and analgesic activities of
aqueous extract of date palm fruit in rats.
Design and Setting: The study was an experimental research carried out at the medical laboratory of University
Sultan Zainal Abidin, Kuala Terengganu, Malaysia.
Methods: Four groups of rats were conveniently assigned with six rats per group based on law of diminishing
return. The groups received different interventions of 500mg of aqueous extract of date palm, 300mg of aqueous
extract of date palm, Paracetamol and normal saline. Hot tail flick test was used to evaluate the analgesic activity
of the rats, while 2.5% croton-oil was used to assess the anti-inflammatory activity in each group.
Results: Statistical significant median difference of the analgesic and anti-inflammatory effect of date fruit
aqueous extract (DFAE) was revealed at (p= 0.001, χ2 =17.84) and (p= 0.002, χ2 =14.80) respectively. In addition,
the significant difference of the analgesic effect of DFAE lies between normal saline and 500mg DFAE (p=0.001,
χ2=-15.250). Likewise the significant difference of the anti-inflammatory effect of DFAE lies between group that
received paracetamol and 300mg DFAE (p=0.027, χ2=0.027), paracetamol and normal saline (p=0.001, χ2=15.717),
and 500mg DFAE and normal saline (p=0.031, χ2=11.467).
Keywords: Date Palm fruit, inflammation and pain.

1. INTRODUCTION

Pain is an unpleasant, emotional and sensory experience as a result of tissue damage or injury (Woolf, 2010). Pain is part
of the inflammatory process (Mironidou-Tzouveleki, 2010). Inflammatory process occurs in order to fight, remove and
repair damaged tissue after injury while pain develops in order to protect the organs from further damage (Warren, 2007).
The other three cardinal signs of inflammation are hotness, redness and loss of function. (Libby, 2007)
Anti-inflammatory conventional drug treatments are limited in their effectiveness in managing the incidence and outcome
of many inflammatory disorders nevertheless NSAIDS and narcotics are the two pharmacological drugs that are available
in the management of pain and inflammation with a serious adverse effect as revealed from a research study (Punchard et
al., 2004). They also present a significant number of side-effects in patients and recently, it has been shown that non-
steroidal anti-inflammatory agents may even attenuate the healing process (Mansouri et al., 2014).

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International Journal of Novel Research in Healthcare and Nursing
Vol. 2, Issue 3, pp: (166-172), Month: September-December 2015, Available at: www.noveltyjournals.com

Natural remedies have been a thing of interest to researchers attempting to find lasting and convincing remedy to diseases
and disorders over the years. Date palm is an ancient plant used in folk medicine for the treatment of various diseases and
disorders (El Hadrami & Al-Khayri, 2012). It is consumed as principal foods and ingredients that form the basis of the
diet of the people of the middle east and the world at large (Al-Juraisy et al., 2010; Friedman et al.,2010). It has been
reported to have high nutritional value and health benefits (Al-Farsi et al., 2007; Sadiq et al., 2013) because it is packed
with essential nutrients, vitamins and minerals that are required for development, growth and overall well being (Ali et al.,
2014; Al-orf et al., 2012). Date palm fruit is a commonly available fruit which is cheap and affordable; being a natural
remedy comes with little or no side effect (Rahmani et al., 2014)
Literature has been reported that date palm posses neuro-protective (Pujari et al., 2011), cerebro-protective (Kalantaripour
et al., 2012), hepato-protective (Ragab et al., 2013) properties, and so much more. This research is primarily based in
finding a remedy with the least or no adverse side effect in the management of pain and inflammation. However in
understanding the anti-inflammatory and analgesic activity of date palm fruit is yet to be known and understood.

2. MATERIALS AND METHODS

Confirmation of date fruit and preparation of aqueous extract:
The date palm was purchased from HIlwah Trading SDN BRD, Kuala Terengganu, Malaysia which has a storage system
which can maintain the quality and hygiene of the fruit. The date was analyzed and confirmed by HERBARIUM UNISZA
with herbarium number 0028.
The pit was separated from the date palm fruit flesh and washed; it was then put into an oven at 500c and allowed to dry.
The dried date palm fruit was then blended in a blender. About 300g of the blended date palm fruit was mixed with
1500ml of normal saline in a conical flask which was then placed in a water bath at 100 0c for 30 minutes. It was then
removed and then passed through a sieve and then through a filter paper. It was then put in a rotary evaporator at 40 0c.
The crude extract was then placed in an oven at 500c and allowed to dry up.300mg/kg and 500mg/kg will be prepared
with normal saline in the ratio 1:2 (v/w).
Drug dosage for required animal:
Doses of extract were selected and determined according to previous study of Elberry and colleagues’ with some
modification in which in their research they use date pollen suspension and extract which show a dependent dose (Elberry
et al., 2011) while in this research date palm fruit aqueous extract was used to see if it will as well produce a dependent
dose.
60ml orange flavored children paracetamol (Smithkline Beecham Philippines) purchased from Farmasi Baiduri Berhad
SDN, Kuala Terengganu Malaysia was mixed with normal saline in fivefold. The syrup contains 50mg per ml of
paracetamol. Therefore the diluted 300ml contain 3000mg of paracetamol. The dosage was calculated according to the
weight of the rat, as the company dosage is 15mg/kg. Normal Saline (0.9% Sodium Chloride, Sigma-Aldrich, USA).
Sample size and design:
This research is an experimental cross sectional research design. Four groups of rats were conveniently assigned with six
rats per group based on law of diminishing return (Charan & Kantharia, 2013). This method is called the resource
equation method. In this method a value E is measured which is the degree of freedom of analysis of variance (ANOVA).
E = Total number of animals − Total number of groups
Group A was given normal saline (0.9%, w/v), (Safi et al, 2011). This served as the negative control. Group B was given
300mg/kg rat of the aqueous extract of date palm fruit by oral gavage. Group C was given 500mg/kg rat of the aqueous
extract of date palm fruit. Group D was given 15mg/kg of paracetamol by oral gavage. This served as the positive control.
They were fed with the extract for three weeks.
Selection of animals, caring and handling:
Sprague-Dawley, 60 days old rats, were housed six per cage in four cages and they were put under controlled condition of
photoperiod controlled room(light: dark: 12h: 12h:), humidity (60+-5%), temperature (22+-2 degrees Celsius), with free
access to food and fresh treated water which free from any harmful impurities. All experiments were conducted in

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Vol. 2, Issue 3, pp: (166-172), Month: September-December 2015, Available at: www.noveltyjournals.com

accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publications
No. 80-23) and were approved by the local ethical committee (University Sultan Zainal Abidin). Approval letter
No.UniSZA.0/3/374-3 (01)
Analgesic effect via hot tail flick test:
The analgesic activity was determined in Sprague dawley rats according to the method of (Rizwan et al., 2003). All drugs
were given orally for 21 days, on the last day the hot tail flick test was done whereby each rat was weighed with its tail
marked about 2cm from the distal end. The rats were put in a restrainer with its tail outside the restrainer, water was
heated at 500C, and kept constant at that temperature, rat tail was inserted into the heated water, the time taken for the rat
to flick its tail or its body sudden jerk due to pain was recorded as the first reading. Time gap for each test was 1min.
reaction time was recorded based on the average of the readings. The maximum time was fixed to 15sec to avoid tissue
damage. The percentage analgesia was calculated using the formula as below
% analgesia=MPE= (TL-BL) / (ML-BL) × 100
Where MPE = Maximum possible effect
ML= Maximum latency or cut off time
TL = Test latency
BL = Basal latency or control latency (Swapnil and Smita, 2012).
Anti-inflammatory effect via croton oil ear edema:
Anti-inflammatory activity was determined according to Florentino and colleagues’ (2013) .The rats were treated with
paracetamol, aqueous extract of date palm fruit and vehicle, for three weeks. The inflammation was induced by the
application of 2.5% (v/v) croton oil solution in acetone (2.5%) on the inner surface of the right ear; the same volume of
acetone was applied to the left ear. After 6 hours the rats were sacrificed and segments of both ears were removed. The
inflammation was measured by the difference between the weights of the segment.(Florentino et al., 2013). The
percentage oedema inhibition was calculated using the formula below
I% = [ 1 – ΔWt /ΔWc] ×100
Where I% = percentage edema inhibition
ΔWt = change in weight of the ear in the treated
ΔWc = change in weight in ear in the control (Boukhatem et al., 2014)
Statistics and data analysis:
The results were analyzed for statistical significance using Kruskall Wallis one way ANOVA; likewise pair wise
comparison test was conducted to determine the significant between group comparisons. A p-value <0.05 was considered
significant. Computer statistical package SPSS (Version 20) was used for analysis.

3. RESULTS
Table 1: Comparison of medial ear weight between four groups and descriptive statistics of %I according to treatments

2
Treatments Dose Median (IQR) %I χ (df) p – value*

Normal saline 3ml 0.13(0.19) --

DFAE 300mg/kg 0.08(0.05) 38 17.84(3) <0.001
DFAE 500mg/kg 0.05(0.02) 61.5
Paracetamol 15mg/kg 0.015(0.05) 88.5
DFAE –Date fruit aqueous extract, I – Edema inhibition *Kruskal-Wallis one way Anova.

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International Journal of Novel Research in Healthcare and Nursing
Vol. 2, Issue 3, pp: (166-172), Month: September-December 2015, Available at: www.noveltyjournals.com

Pair wise comparison test with level of significant set at 0.05, shows that the significant difference lies between group
that received paracetamol and 300mg DFAE (p=0.027,χ2=0.027), paracetamol and normal saline (p=0.001, χ2=15.717),
and 500mg DFAE and normal saline (p=0.031, χ2=11.467),

Croton oil induce injury

100
90
80
70
60
weght(g)

50
40
30
20
10
0
Group A Group B Group C Group D
Median (IQR) 0.13 0.08 0.05 0.015
%I 0 38 61.5 88.5

Fig 1: Determination of %I using croton oil induce edema

Table 2: Comparison of tail flick time between four groups and descriptive statistics of %analgesia according to treatments

2
Treatments Dose Median (IQR) %A χ (df) p – value*

Normal saline 3ml 2.0(0.25) --

DFAE 300mg/kg 3.5(1.75) 11.54 14.80(3) <0.002

DFAE 500mg/kg 6.5(2.75) 34.62

Paracetamol 15mg/kg 4.0(2.00) 16.67

DFAE –Date fruit aqueous extract, A - analgesia, *Kruskal-Wallis one way anova test.

Pair wise comparison test , with level of significant set at 0.05, shows that the significant difference lies between normal
saline and 500mg DFAE (p=0.001, χ2 = -15.250)

40
Tail flick time
35
30
25
time(secs)

20
Median
15 (IQR)
10 %MPA
5
0
Group A Group B Group C Group D
Groups

Fig 2: Determination of %analgesia using tail flick test

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4. DISCUSSION
Pre treatment with 500mg of aqueous extract of date palm fruit resulted in dose dependent reaction in croton oil evoked
ear edema and differed significantly (P<0.001) among different groups of rats (table 1 and fig 1). The result showed that
significant difference lies between group of paracetamol and 300mg (p=0.027), group of paracetamol and normal saline
(p=0.001) and group of 500mg and normal saline (p=0.031). The descriptive statistics show that paracetamol tend to
reduce the inflammation by 43.9% higher than 500mg, 132.9% higher than 300mg while 500mg tend to reduce the
inflammation by 61.8% higher than 300mg however 500mg produce a dependent dose. There is every possibility with
increase dose of the extract % inhibition can be more than that of paracetamol.
This is in line with the study done by (Mohamed & Al-Okbi, 2004) on the anti-inflammatory effect of 500mg methanolic
and water of edible date palm fruit and 500mg of methanolic extract of date palm seed on adjuvant arthritis, which reveals
that oral administration of both extract reduce the paw inflammation as well as the swelling throughout the experiment.
Likewise it is in line with the study by Elberry and colleagues as they evaluate the anti-inflammatory effect of date palm
on ATP which shows that date palm possesses anti-inflammatory activity as it reduce the size of the anterior and posterior
lobes of the prostate as well as decreasing the proinflammatory cytokines in an atypical prostatic hyperplasia (ATP)
induce rat (Elberry et al., 2011). These finding also support the finding of Mukherjee et al., (2014) as they reported that
date palm possesses not only anti-inflammatory effect but also anti-oxidant effect. Research have shown that this effect
could be due to the present of antioxidant in date palm (Mukherjee et al., 2014).
Pretreatment with 500mg of date palm fruit aqueous extract resulted in dose dependent reaction in hot tail flick test and
differed significantly (P<0.002) among different groups of rats (table 2 and fig 2). The result shows that the significant
difference lies between normal saline and 500mg DFAE (p=0.001). Likewise, the descriptive statistics shows that the
%analgesia of 500mg is 200.2% higher than 300mg, 108.4% higher than paracetamol, while paracetamol was 44.2%
higher than 300mg.
This findings support the study of Alkuran and colleagues as they evaluated the effect of consuming date palm fruit on
pregnant women which showed that women who consume date palm fruit four weeks prior to delivery tend to have ease
pain during labor as it reduced the pain of labor (Al-kuran et al, 2011). This is also in line with a study done by shabani
and colleagues in which they investigated the effect of date extract consumption on thermal hyperalgesia of neuropathic
pain in diabetic rats which showed that date extract can not only decrease the thermal hyperalgesia but also can prevent
pain resulting from diabetic neuropathy(Shabani et al., 2013).

5. CONCLUSION
The outcome of the study revealed that 500mg of DFAE is effective in the reduction of pain than 300mg and paracetamol
while paracetamol tend to inhibit edema more as compared to 500mg and 300mg but the percentage edema inhibition of
paracetamol is comparable to that of 500mg of the extract. The effectiveness of the extract could be due to the presence of
micronutrients in the date palm fruit especially vitamin C and E as compared to paracetamol, as well as due to the
presence of antioxidants present in the fruit as compared to paracetamol. It can be concluded that feeding or oral
administration of aqueous extract date palm fruit in high dose proves to be more effective in the reduction of pain and
inflammation of an acute injury in rats. Being the available analgesic and anti-inflammatory drugs are associated with
serious adverse effects; this research can bring another option to develop safer free from adverse effect and more effective
drug in the management of pain and inflammation. By determining the histological severity of the injury can improve and
provide deeper understanding of pathophysiology of pains as well as shade more light on the safety and efficacy of the
drug candidate and hence develop better pain management modalities and approaches in the future.

ACKNOWLEDGEMENT
I wish to thank the Kano State Government of Nigeria and my husband Auwal Muhammad Abdulkareem for the courage
and support in the smooth conduct of this study. Likewise, to my humble supervisor Professor Nordin bin Simbak for all
the corrections given during the course of the study.

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REFERENCES
[1] Al-Farsi, M., Morris, A., & Baron, M. (2007). Functional properties of omani dates (Phoenix dactylifera L.). In Acta
Horticulturae.
[2] Ali, A., Waly, M., Essa, M. M., & Devarajan, S. (2014). Nutritional and Medicinal Value of Date Fruit.
[3] Al-Juraisy, Y., Yaseen, N. Y., Al-Ani Professor Emeritus, B., & Dept, B. (2010). (Phoenix dactylifera L. cv. Zahdi)
on some cancer cell lines in vitro and treatment of transplanted mammary adenocarcinoma in mice Effect of crude
extracts of fruits and pits of date palm.
[4] Al-orf, S. M., Ahmed, M. H. M., Atwai, N. A.-, Al-, H., Dehwah, A., Dehwah, S., … Arabia, S. (2012). Review:
Nutritional Properties and Benefits of the Date Fruits ( (Vol. 2012).
[5] B.H Rizwan, T. C. L. and S. Z. I. (2003). Ridwan, the antinoceeptive effects of water extracts from sea
cucumbers.pdf. Pakistan Journal of Biological Science, 6(24), 2068–2072.
[6] Boukhatem, M. N., Ferhat, M. A., Kameli, A., Saidi, F., & Kebir, H. T. (2014). Lemon grass ( Cymbopogon citratus
) essential oil as a potent anti-inflammatory and antifungal drugs. Libyan Journal of Medicine, 1(9), 1–10.
[7] Charan, J., & Kantharia, N. D. (2013). How to calculate sample size in animal studies? Journal of Pharmacology &
Pharmacotherapeutics, 4(4), 303–6. http://doi.org/10.4103/0976-500X.119726
[8] El Hadrami, A., & Al-Khayri, J. M. (2012). Socioeconomic and traditional importance of date palm. Emirates
Journal of Food and Agriculture.
[9] Elberry, A. A., Mufti, S. T., Al-Maghrabi, J. A., Abdel-Sattar, E. A., Ashour, O. M., Ghareib, S. A., & Mosli, H. A.
(2011). Anti-inflammatory and antiproliferative activities of date palm pollen (Phoenix dactylifera) on
experimentally-induced atypical prostatic hyperplasia in rats. Journal of Inflammation. http://doi.org/10.1186/1476-
9255-8-40
[10] florentino F. Iziara, marcus vinicius m. Nascimerto, Publinny m. Galdino, Adriane F De Breto, Fabio F Da Rocha,
Carlos R Tonussi, Thereza Christina MDe Lima, jose R. D. and E. A. costa. (2013). Evalution of analgesic and anti-
inflammatory activities of hydrocotyle umbellate L.. Araliaceae (acaricoba in mice. An Acad Bra Scienc, 85(3).
[11] Friedman, M. H., Andreu, M. G., Quintana, H. V, & Mckenzie, M. (2010). Phoenix dactylifera, Date Palm 1.
[12] Swapnil, J.R and S. D. Smita. (2012). Experimental evaluation of analgesic and anti-inflammatory activity of
simvastatin and atorvastatin. Indian Journal of Pharmacology, 44(4), 475–479.
[13] Kalantaripour T.P., Asadi-shekaari M., B. M. and N. G. . (2012). cerebroprotective. Journal of Biological Science.
http://doi.org/10.3923/jbs.2012
[14] Libby, P. (2007). Inflammatory Mechanisms: The Molecular Basis of Inflammation and Disease. Nutrition Reviews.
http://doi.org/10.1111/j.1753-4887.2007.tb00352.x
[15] Mansouri, L. El, Bousta, D., Boukhira, S., Balouiri, M., Khanchoufi, A. El, Akdime, H., … Achour, S. (2014).
International Journal of Phytopharmacology. International Journal of Phytopharmacology, 5(2), 76–84.
[16] Mironidou-Tzouveleki, M. (2010). Inflammation and pain. http://doi.org/10.1186/1744-859X-9-S1-S2
[17] Mohamed, D. A., & Al-Okbi, S. Y. (2004). IN VIVO EVALUATION OF ANTIOXIDANT AND ANTI-
INFLAMMATORY ACTIVITY OF DIFFERENT EXTRACTS OF DATE FRUITS IN ADJUVANT ARTHRITIS.
POLISH JOURNAL OF FOOD AND NUTRITION SCIENCES Pol. J. Food Nutr. Sci, 1354(4), 397–402.
[18] Mukherjee, K., Paul, P., & Banerjee, E. R. (2014). Free Radical Scavenging Activities of Date Palm (Phoenix
sylvestris) Fruit Extracts. Nat Prod Chem Res, 2. http://doi.org/10.4172/2329-6836.1000151
[19] O. Al-kuran, L. Al-mehaisen, H. Bawadi, S. B. and Z. A. (2011). The effect of late pregnancy consumption of date
palm fruit on labour and delivery. Journal Obstet Gynaecol, 31(1), 29–31.

Page | 171
Novelty Journals
 ISSN 2394-7330
International Journal of Novel Research in Healthcare and Nursing
Vol. 2, Issue 3, pp: (166-172), Month: September-December 2015, Available at: www.noveltyjournals.com

[20] Pujari, R. R., Vyawahare, N. S., & Kagathara, V. G. (2011). Evaluation of antioxidant and neuroprotective effect of
date palm (Phoenix dactylifera L.) against bilateral common carotid artery occlusion in rats. Indian Journal of
Experimental Biology, 49, 627–633.
[21] Punchard, N. A., Whelan, C. J., & Adcock, I. (2004). The Journal of Inflammation. Journal of Inflammation
(London, England), 1(1), 1–4. http://doi.org/10.1186/1476-9255-1-1
[22] R. Ragab, Ahmed, Elkablawy A. Mohammad, B. Y. S. and H. N. B. (2013). Antioxidant and Tissue-Protective
Studies on Ajwa Extract: Dates from Al-Madinah Al-Monwarah, Saudia Arabia. Journal of Environmental &
Analytical Toxicology, 3(163), 1000163. http://doi.org/10.4172/2161-0525.1000163
[23] Rahmani, A. H., Aly, S. M., Ali, H., Babiker, A. Y., Suikar, S., & Khan, A. A. (2014). Therapeutic effects of date
fruits (Phoenix dactylifera) in the prevention of diseases via modulation of anti-inflammatory, anti-oxidant and anti-
tumour activity. International Journal of Clinical and Experimental Medicine, 7(3), 483–491.
[24] Sadiq, I. S., Izuagie, T., Shuaibu, M., Dogoyaro, A. I., Garba, A., & Abubakar, S. (2013). The Nutritional Evaluation
and Medicinal Value of Date Palm (Phoenix dactylifera). International Journal of Modern Chemistry International
Journal of Modern Chemistry Journal Homepage: www.ModernScientificPress.com/Journals/IJMChem.aspx Int. J.
Modern Chem, 4(43), 147–154.
[25] Shabani, M., Zangiabadi, N., & Asadi-Shekaari, M. (2013). Evidence for Positive Effects of Date Extract That
Attenuates Thermal Hyperalgesia in a Diabetic Rat Model of Neuropathic Pain. Neuroscience & Medicine, 4, 16–22.
http://doi.org/10.4236/nm.2013.41003
[26] Warren, D. K. (2007). Core Topics in Pain. Anesthesia & Analgesia. http://doi.org/10.1213/01.ANE. 0000251473.
15241.A9
[27] Woolf, C. J. (2010). What is this thing called pain? Journal of Clinical Investigation, 120(11), 3742–4.

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