Phoenix dactylifera L.

DOI: 10.17354/ijpphs/2015/20

Original Article

Evaluation of Total Flavonoid Contents of Mariami Iranian Date Fruits (Phoenix dactylifera L.) using Different Extracts Ibrahim H Sani1, R Mohammed Adzim Khalili2, U Abdulkareem3 Post-graduate Student, Department of Addiction Medicine, Faculty of Health Sciences, Universiti Sultan Zainal Abidin, Habsah Block, Gong Badak Campus, 21300 Kuala Terengganu, Terengganu, Malaysia, 2Medical Lecturer, Department of Addiction Medicine, Faculty of Health Sciences, Universiti Sultan Zainal Abidin, Habsah Block, Gong Badak Campus, 21300 Kuala Terengganu, Terengganu, Malaysia, 3Post-graduate Student, Department of Neuroscience, Faculty of Health Sciences, Universiti Sultan Zainal Abidin, Habsah Block, Gong Badak Campus, 21300 Kuala Terengganu, Terengganu, Malaysia 1

ABSTRACT Introduction: Phoenix dactylifera Linn., a date palm is known for its nutritional and numerous health benefits due to its presence of bioactive compounds. Iranian Mariami date fruit (IMDF) is widely found and used by people around the globe. Purpose: This study was aimed to determine the total flavonoid contents (TFCs) presence in IMDFs of the three different extracts (methanolic extract, hydro-ethanolic extract, and aqueous). Materials and Methods: All the samples were extracted from three different solvents and measured using aluminum chloride colorimetric method, with some modification. Result: The TFCs of the extracts in decreasing order was methanolic extract (11 mg quercetin [QE]/g) > hydro-ethanolic extract (7.1 mg QE/g) > aqueous (6.8 mg QE/g), respectively. Conclusion: The antioxidant potential effect of IMDFs was due to its high TFCs, which can be benefited to human body systems. In this study, methanol solvent shown to have a high affinity of extracting the flavonoid contents. Key words: Antioxidant, Flavonoids, Health, Phoeniceae, Plants

INTRODUCTION Phoenix dactylifera Linn. (PD) (Date palm) that belong to family Arecaceae, called “Nakhla,” the “Tree of Life” by the Arabs, is considered as most popular cultivated product plants in the Middle East since at least 6000 BC, and it is accepted to be native to the countries around the Arabian Gulf.1 Many Middle Easterners believe that eating of date fruits has a lot of medicinal benefits. Different parts of this plant are traditionally demanded to be used for the treatment of a broad spectrum of disease conditions including memory disturbances; there has been a great interest in this plant as evidenced by the tremendous work in last few decades.2 Date palm fruits (DPFs) have been an important component of natural sources of food nutrients in most of the regions of the world and high-energy food rich in carbohydrates, including dietary fiber and minerals, for example, potassium, magnesium, calcium, phosphorus, and iron. Most of the carbohydrates in dates are in the form of fructose and glucose, which are readily

absorbed by the human body.3,4 Muslims all over the world break their fasting during the month of Ramadan by taken date fruits.5 Iran, one of the date palm producing countries with more than 400 varieties that pretty nearly 185,000 ha of its properties is under date fruits cultivation.6 However, recent studies show that consumption of fruits and vegetables reduces the risk of several chronic diseases such as cardiovascular diseases, neurological diseases, obesity, and cancers.7 The natural antioxidant products, which neutralize free radicals, have been known to have more attention by scientists for their potential impacts to prevent and control neurodegenerative and chronic disease.8 However, the antioxidant can be as any substance that present at low concentrations will significantly delay or prevents oxidation of the substrate; it was found that presence of flavonoid compounds in the PD fruits is efficient scavengers of free radicals.9 Furthermore, the natural products continue to provide valuable therapeutic agents, both in modern and in traditional

CORRESPONDING AUTHOR: Dr. Ibrahim H Sani, Universiti Sultan Zainal Abidin, Gong Badak Campus, 2130 Kuala Terengganu, Terengganu, Malaysia. Mobile: (+60)168067630/(+234)8036858682. E-mail: [email protected] Submission: 08-2015; Peer Review: 09-2015; Acceptance: 10-2015; Publication: 10-2015;

International Journal of Preventive and Public Health Sciences • Sep-Oct 2015 • Vol 1 • Issue 3

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Sani, et al.: Evaluation of Total Flavonoid Contents of Iranian Mariami Date Fruits using Different Extracts

medicine.10 The various parts (fruit, seed leave, and pollen grain) of PD are widely used in the traditional medicine for the treatment of numerous disorders which include memory disturbances, fever, inflammation, paralysis, loss of consciousness, nervous disorders, and it is also scientifically proved to possess pharmacological activities which demonstrate its benefits in different sorts of disorders and disease condition.11 The high nutritional values of dates are also as a result of their dietary fiber content, which makes them suitable for the preparation of fiber-based foods and dietary supplements. Dietary fiber has important therapeutic implications for certain conditions such as obesity, hyperlipidemia, and diabetes, it exhibit a protective effect against hypertension, coronary heart disease, anticancer, antioxidant, hepatoprotective, antiulcerative, anti-inflammatory, antiproliferative, antimutagenic, antibacterial, antifungal, and antiviral potential.1,12,13 Moreover, the date palm has been used for prevention and treatment of various disease conditions and healthy individuals. Studies reported that an antioxidant activity of PD is as a result of attribution of flavonoids (flavones, flavonols, and flavanones).4,14,15 Flavonoids are polyphenolic compounds that are abundant in nature and are classify, by their chemical structure, into flavones, flavonols, isoflavones, flavanones, chalcones, anthocyanidins, and catechins or as per their molecular structure that comprises two benzene rings joined by a straight three-carbon chain and structures an oxygenated heterocycle (C6-C3-C6) and their vast number emerges from the different combinations of multiple hydroxyl, methoxyl, and O-glycoside bunch substituents on the essential benzo-pyrone (C6-C3-C6).16 However, its believed to be one of the most common components in the human diet, the average intake of all flavonoids is evaluated to be 1 g/day.17 In fact, flavonoids present in plants possess various health benefits, such as antioxidant activities; reduce the risk of certain chronic diseases, cardiovascular disorders, and certain kinds of cancerous processes.18 Flavonoids reported to possess excellent antioxidant properties related to their rules to delay/interfere with the development and production of free radicals and protect low-density lipoproteins from oxidation.19 However, in the central nervous system (CNS), flavonoid exert it effect either by acting as inhibitors of monoamine oxidase A or B, subsequently serving as anti-depressants or anti parkinsonism’s or by binding to the benzodiazepine site on the gamma-aminobutyric acid (GABA)-receptor resulting in sedation, anxiolytic, or anti-convulsive effects.20,21 A recent studies suggested that flavonoids served as prominent agents in the treatment of neurological/mental disorders, the neuro-protective actions of natural flavonoids involved significant effects within the brain, including attenuation of oxidative stress, a potential protection of neurons against injury induced by neurotoxins, their capacity to suppress the neuro-inflammation, and the possibility to promote cognitive function, learning and memory.22,23 Another reported suggested that flavonoids increase endothelial nitric oxide production resulting in endothelial relaxation in conditions such as high blood pressure, metabolic syndrome, and stroke.24 However, shown significant effects of biochemical and pharmacological actions which are indicative of anti-inflammatory effects, modulate the function of inflammatory cells such as lymphocytes, natural killer cells, monocytes, neutrophils, mast cells and macrophages, and modulation of the immune system.25 Furthermore, it exerts 30

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health benefits to respiratory and digestive systems by acting as anti-allergic, antioxidant, and antispasmodic actions which are beneficial in respiratory tract diseases and inhibit gastrointestinal motility and diarrhea.20 PD L is well known for its nutritional value and numerous health benefits due to its presence of bioactive compounds. It is, therefore, of great important to assess the total flavonoid contents (TFCs) of Iranian Mariami date fruits (IMDFs). Thus, a lot of researches has been carried out on PD, which pay more attention to the antioxidant activity of most popular date fruits such as Ajwa. However, this current study was aimed to determine the TFC presence in IMDF of the three different solvent extracts.

MATERIALS AND METHODS Chemicals and Reagents Methanol, ethanol, analytical grade distilled water, quercetin (QE), sodium carbonate, and aluminum chloride. All chemicals were purchased from Sigma-Aldrich (Malaysia). Extraction of the Fruit An IMDF (PD) was purchased; date fruit was purchased from a local supermarket, Terengganu, Malaysia. The fruit was authenticated by a Botanist at Faculty of Biotechnology Universiti Sultan Zainal Abidin with the voucher number: 00289. The fruits were washed and separated into pit and flesh, from a local supermarket, Terengganu, Malaysia. The flesh was further cut into pieces, dried in an oven and blended. 100 g of blended fruit was weighed and placed inside 1000 ml conical flask, macerated with 500 ml of test solvents, and placed for 48 h in a dark cupboard with vigorous shaking at intervals. The mixture was then filtered using Whatmann No. 4 filter paper, and the filtrate was concentrated using rotatory evaporator. The concentrated extracts were then placed in the oven at 40°C to allow for complete evaporation. All the three extracts were stored at −20°C in a refrigerator. Determination of Total Flavonoids Contents The TFC was determined by aluminum chloride colorimetric method, described by Abuelgassim,26 with some modification. Briefly, 0.5 ml of 1 mg/ml of extracts was mixed with 0.5 ml of distilled water, followed by 0.1 ml of 10% aluminum chloride, 0.1 ml of 1 M potassium acetate, and 2.8 ml of ethanol (95%) were added in order. After incubation at room temperature for 40 min, the absorbance was measured at 415 nm wavelength. A calibration curve was prepared using a standard solution of QE (20, 40, 60, 80, 100, 120, 140, and 160 µg/ml, R2 = 0.9963). The TFC in samples was expressed as milligram QE antioxidant equivalents (QAE)/gram of extract (Figures 1 and 2a and b). Data Analysis The data are presented as means ± standard deviation. All samples were analyzed in three replications.

RESULTS The standard QE calibration curve of total flavonoid with the QE as a reference standard is presented in Figures 3 and 4. The outcome from Table 1 revealed a higher TFC in IMDF, based on the three studied samples. Regression equation from standard QE calibration curve: Y= 0.0052x + 0.0883, R2 = 0.9963 and DME with the highest while DA with the lowest TFC.


www.ijpphs.comSani, et al.: Evaluation of Total Flavonoid Contents of Iranian Mariami Date Fruits using Different Extracts

Table 1: The percentage yield of three different extracts of IMDF Extracts

Weight of samples (g)

Volume of solvents (ml)

TFC mg/g of extract

DME

100

500

11.0

DEW

100

500

7.1

DA

100

500

6.8

100 g of sample soaked in 500 ml of solvents (methanol, hydro‑ethanol, and water). DME: Methanolic extract, DEW: Hydro‑ethanolic extract, DA: Aqueous extract, IMDF: Iranian Mariami date fruit

DISCUSSION

Figure 1: The brief explanation of extraction process

a

b

Figure 2: (a and b) Reading of the absorbance using spectrophotometer

Figure 3: Standard quercetin (QE) calibration curve of QE, with linearity of regression equation as Y = 0.0052x + 0.0883, R2 = 0.9963

The protective role of flavonoids in natural products, to various human body systems has been pointed out.20,21,24 Therefore, in this study, we determined the TFC of IMDF that is one of the most commonly used date fruits in Malaysia. The standard calibration curve of total flavonoid with the QE as a reference standard is presented in Figures 1 and 2. The TFC of IMDF were found to be 11.0, 7.1, and 6.8 (mg QE/g) in DME, DEW, and DA, respectively. The DME showed the highest concentration in total flavonoid with 11.0 mg QE/g, followed DEW and DA as shown in Figures 1 and 2. Similar reported that date palm leaves with the highest TFC among other plants 11.55 mg QAE/g higher than what obtained in this study.26 However, higher concentration obtained in the date palm leave than the concentration of total flavonoid in the DPF, which is reported to be 81.79 mg CE/100 g (catechin equivalents).27 Two similar studies evaluated the antioxidant activity of Saudi date palm (PD) fruit of various cultivars with the TFC range from 1.44 to 3.94 mg QE/100 g DW and 310.3554.0 (mg CE/100 g of syrup), respectively, the outcome was lower compared to the present study.28,29 Similarly, the study revealed that the TFC of methanolic extract of Deglet-Nour Date to be 2.34 mg QE/100 g fresh weight, which is lower, compared to the outcome of this study.7 In contrast, the outcome of the study carried out in Saudi and Egypt revealed a higher TFC of 17.69 ppm and 54.06 ppm (mg/L), respectively, compared to the present result.30 In fact, this current study has shown high TFC of IMDF compared to other varieties of date fruits. Furthermore, considering various health benefits of the flavonoid compound which include antioxidant and radical scavenging activities. Reduces the risk of cardiovascular, chronic diseases, and certain kinds of cancerous processes.18 The variation in the outcome of this study, compared to the previous study might be as a result of the type of the date fruits and the method used for extraction. The role of flavonoid in the CNS can best be explained by acting as inhibitors of monoamine oxidase A or B, thereby working as anti-depressants or anti parkinsonism’s or by binding to the benzodiazepine site on the GABA-receptor, which give rise to its sedating, anxiolytic, and anti-convulsive effects.20,21,23 Public Health Significance

Figure 4: Total flavonoid contents as quercetin equivalent (QE mg/g). DME: Methanolic extract, DEW: Hydro-ethanolic extract, DA: Aqueous extract

This study supported previous research that flavonoids served as prominent agents in the treatment of neurological disorders, the neuro-protective actions of natural flavonoids involved a significant effects within the brain, including attenuation of oxidative stress, a potential protection of neurons against injury induced by neurotoxins, their capacity to suppress the neuroinflammation, and the possibility to promote cognitive function, learning and memory.


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Sani, et al.: Evaluation of Total Flavonoid Contents of Iranian Mariami Date Fruits using Different Extracts

CONCLUSION The outcome of this study revealed that IMDFs have a promising antioxidant potential due to its high TFC, which can be benefited to human body systems. Thus, regular intake of date fruits is beneficial to our body systems.

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HOW TO CITE THIS ARTICLE: Sani IH, Khalili RMA, Abdulkareem U. Evaluation of Total Flavonoid Contents of Mariami Iranian Date Fruits (Phoenix dactylifera L.) using Different Extracts. Int J Prevent Public Health Sci 2015;1(3):29-32.
