Free Radical Scavenging Potency of Five Date Varieties at Different Stages by Using DPPH Assay
Received Date: May 16, 2018 / Accepted Date: Jun 04, 2018 / Published Date: Jun 21, 2018
Samples of date fruit was collected from Khairpur Mir’s district and examined for antioxidant activity, for this purpose extract was prepared in different diluents like Methanol+Water (50:50), methanol and water by using DPPH assay. It was observed from data that all the extract have impressive antioxidant ability against free radicals. Among them solution have more ability to ramp the free radicals. From data, it was concluded that Gajar Wari is more active as it possess lowest IC50 value (20.022 μg/mL in Methanol) then rest of other date varieties at final stage in solution as well as Aseel variety also possesses lowest IC50 value (18.33 μg/mL), there for have more ability against free radicals in methanol at second stage.
Keywords: Antioxidants; DPPH; Date fruit; Methanol; Water
The species which have ability to neutralize free radical by donating an electron is known as antioxidant [1-3] and reduce its ability to damage vital molecules. Antioxidant plays a vital role in human health therefore it gets more importance in medical field and also in the field of food industry . Free radicals can damage vital bioactive molecules such as DNA and Protein and generate many chronic and cardiovascular diseases [5-7]. While human body retains such method of shield, as enzymes and antioxidant nutrients, which capture harmful possessions of Reactive Oxygen Species (ROS), constant mixing to chemicals and pollutants might promote the concentration of ROS in body behind its ability to hold such things, causing irreparable oxidative harm. Hence, antioxidants might have been important application in the inhibition and have healing power of disorder wherein free radicals are concerned [8,9]. Plants, fruits and vegetables are primary source for polyphenolic compounds such as phenolic acids and flavonoids  that are known as natural antioxidants that possess many biological effects and have ability to decrease risk of diseases. Date fruit contains many essential minerals . It also possesses protein and amino acids in higher concentration than other fruits [12-14]. Besides its nutritional value, the fruit of date palm are full of phenolic compounds owing antioxidant ability against free radicals [15-17].
Free radical rummaging potency was carried out via DPPH assay procedure that is very simple and easy method which has been founded on decreasing of DPPH, a stable free radical [18,19]. Due to the number of odd electrons free radical DPPH produce extreme absorption at 517 nm (purple color) . Now antioxidants can interact with 1, 1-diphenyl-2-picrylhydrazyl and free radical has been balancing in the existence of a hydrogen contributor (e.g., a free radical scavenging antioxidant) and convert to the DPPH-H and as significance the absorbance’s reduced from the DPPH. The formation of DPPH-H from radical outcomes decolorization of color which is associated through quantity of electrons trapped . The reducing ability depends on decolorization. In this way analysis is being recognized perfect for assessing oxidants rummaging activity of somewhat novel medication. Although DPPH solution to be mingled by means of material that can be able to contribute a hydrogen atom, in this way it promotes to the diminish form (Diphenyl picryl hydrazine; non radical) by the losing the lilac color . From the results it was reported that fruit of this plant possesses antioxidant properties.
Materials and Methods
The sampling of date fruit was carried out by international scientific methods of sampling as given in AOAC  and varieties of date fruit was identified with the help of growers followed by botanist. However, stages of fruits were differentiating and classified on the basis of color as mentioned in literature . The five fresh date ripening varieties such as Karbala Wari, Kupro, Gajar Wari and Asul Khurmo were collected at their final ripening stages while date variety Aseel were collected at their different ripening stage during the summer of 2014 from the district Khairpur. Screening of samples was done randomly two times at their different fully grow period in three replicates and packed in polyethylene bags exception chemical treatment for the period of two hour, finally samples were brought into laboratory Institute of chemistry, Shah Abdul Latif University, Khairpur for free radical scavenging potency.
A portion of date sample 0.025 g was disintegrated in 25 mL of different solvents such as water, methanol and methanol+water (50:50) afterward mingled through magnetic stirrer to half an hour it was being purified with filter paper.
A DPPH was prepared as stock solution. For this regard 164 μM methanolic solution was prepared in 50 mL conical flask which is coated with aluminum foil in dark to avoid the oxidation [24,25]. Absorbance was recorded to check the stability of radicals.
Free radical scavenging potency was accomplished through consuming 1,1-diphenyl-2-picrylhydrazyl spectrophotometric method [26,27], about addition 1 mL of DPPH solution is combined to 2 mL of extract of distinct solvents to prepared 3 mL solution along with blank and this solution was permitted to stand at room temperature in the absence of light for 30 min. The measurement of absorbance was performed at 517 nm using spectrophotometer ((UV-Vis Shimadzu) . Ascorbic acid was used as standard solution. Every test was conceded out in triplicates and the following formula has been used for calculation of percent inhibition:
Here Ao stands for blank absorbance of Blank and as stands for sample absorbance.
The value of IC-50 is to be the concentration of sample essential to prevent 50% of the DPPH free radical was intended by means of Log dose inhibition curve [29,30]. The samples with lower absorbance of the reaction combination indicated more ability to ramp the free radical ability [31,32].
Results and Discussion
From the Table 1, it is was detected that date variety Aseel displays 28.06 μg/mL, 18.33 μg/mL, 35.23 μg/mL and 22.32 μg/mL in methanol, IC50 in water was measured as 23.79 μg/mL, 24.40 μg/mL, 20.87 μg/ mL and 22.62 μg/mL while in methanol : water IC50 value was measure such as 25.12 μg/mL, 22.14 μg/mL, 26.61 μg/mL and 23.81 μg/mL at 1st, 2nd, 3rd and 4th stags respectively, from above data it is clear that Aseel possesses lowest IC-50 value in methanol (18.33 μg/mL) which mirrors that extracts in methanol solvent possesses more ability against free radical with respect to other date varieties.
Note: (IC-50 value in μg/mL)
Table 1: Comparative antioxidant potency of Aseel at different stages in different solvents.
Similarly, from the Table 2, it was observed that IC-50 value of five date varieties at final stage measures as Aseel have 22.32 μg/mL, 22.62 μg/mL and 23.81 μg/mL, Gajar Wari shows 20.02 μg/mL, 20.32 μg/mL, and 27.83 μg/mL Karbala Wari 22.10 μg/mL, 23.95 μg/mL and 23.54 μg/mL, in Kupro found as 23.11 μg/mL, 28.35 μg/mL and 20.83 μg/mL while Asul Khurmo has 27.34 μg/mL, 25.77 μg/mL and 25.35 μg/mL in methanol, water and methanol:water respectively at 4th stages.
Note: (IC-50 in μg/mL)
Table 2: Relative free radical strength of five date varieties at final stages in different solvents.
With accordance of experimental study by enhancing amount of sample, absorption decreased while % inhibition enhanced. Huge absorption denoted higher number of free DPPH molecules [17,33] whatever diminished absorbance displayed combination of free DPPH molecules with amount of sample. By using lower Concentration, the IC50 value was to be gained less and greater ability to ramp the free radical against oxidants (Figures 1 and 2) .
It is observed from the data that at Tamar stage all the five date varieties have better capability in contradiction of oxidation but date variety Gajar Wari has lesser IC-50 value that is 20.02 μg/mL in Methanol as compared to further date varieties and have the great antioxidant ability to ramp oxidants. In water Kupro possesses IC50 value 28.35 μg/mL at Tamar stage and holds less antioxidant capacity.
Similarly, date variety Aseel in all solvents shows great antioxidant ability, among these solvent in Methanolic solution at 2nd stage less IC50 value observed that is 18.33 μg/mL and also minimum value observed (35.23 μg/mL). It has been measured that lesser the IC-50 value is related with little quantity of sample while antioxidant capacity will be superior.
This research work was carried out in the Institute of Chemistry, Shah Abdul Latif University Khairpur Mir’s Pakistan. The authors warmly express thanks and gratefully acknowledge the assistance and moral support provided and expresses their appreciations for the provision of necessary facilities.
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Citation: Mahar ZA, Shar GQ, Lal B, Solangi ZA (2018) Free Radical Scavenging Potency of Five Date Varieties at Different Stages by Using DPPH Assay. J Nutr Food Sci 8: 707. DOI: 10.4172/2155-9600.1000707
Copyright: © 2018 Mahar ZA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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