In Vitro Antioxidant Property of Two Syzygium Species Leaf Extract

Published: 2021-07-11 20:50:05
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Antioxidants are compounds that protect cells on the effects of free radicals- highly reactive, unstable particles that are generated during normal cellular processes or when the body is exposed to radiation, intense physical exertion, or tobacco smoke. These powerful compounds keep errant oxidation under control by giving electrons to the free radicals, thus neutralizing these particles. However, free radicals in the human body outnumber the naturally occurring antioxidants which makes it necessary to have a continuous intake of external sources of antioxidants.
Fruits, vegetables, and whole grains are good external sources of antioxidants such as vitamins C and E, and carotenoids. On the other hand, naturally occurring antioxidants include phytochemicals such as lignans, tannis, phenols, and flavonoids. These are substances that can be found in plant-based foods.Moreover, plant-based food like Makopa and Tambis, are not known for its medicinal properties specially in the Philippines because researches about these two Syzygium species are uncommon. These are Syzygium species that are abundant in the Philippines. In the present study, essential oil yielded from fresh leaves of Syzygium species like Makopa and Tambis showed monoterpenes characterized mainly by α-pinene, β-pinene, p-cymene, and α-terpineol. These phytochemicals are classified as terpinolene- an organic compound present in the essential oils obtained from leaves. It has therapeutic attributes namely anticancer, antibacterial, antioxidant, antibacterial, and sedative properties. Also, terpinolene exhibit high antibacterial activity against common bacteria like Staphylococcus epidermidis, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, and Pseudomonas aeruginosa.
Despite having these therapeutic attributes, Makopa and Tambis remain to be unfamiliar in the field of medicine. To maximize the benefits from these Syzygium species, the researchers aims to explore the possibilities of two Syzygium species leaf extracts as a source of external antioxidants for the human body. Makopa and Tambisare easily obtainable, thus making it affordable. Furthermore, this research would be a great help in the discovery of natural external sources of antioxidants and fresh perspective on the unpopular, beneficial properties of Makopa and Tambis.
The research identified the certain phytochemicals of the Makopa and Tambis leaf extract are considered to have antioxidant properties. The study provided the leaves of the two Syzygium species another use, besides the utilization of the fruit itself in different areas of medicine. This study might also provide information to some people regarding the other uses of Syzygium species, in that their purpose is not solely for eating, but the other parts can also be utilized in the field of medicine. It would be beneficial to pharmaceutical industries to use this research for the utilization of the two Syzygium species for production of medicine derived from its properties.
Also, it would be helpful to farmers of this plant species because this research would discover other uses for their plant. Furthermore, the research may be of service to future researchers because it would help them in learning about the properties of the Syzygium species, not only of the fruit, but also of the other parts of the plant like the leaves.
The main focus of this research was to determine the anti-oxidant properties of Syzigiummalaccense leaf extract and Syzigiumaqueum leaf extract through DPPH radical scavenging assay. Only one parameter was tested which was the amount of concentration of the leaf extracts. Testing was done outside the school premises with the help experts because of the absence of facilities in the campus. Definition of termsIn this research study, the following terms were operationally defined to improve the understanding of this work. DPPH assay. A bioassay test offered in University of the Philippines- Manila that is used to evaluate the antioxidant property of Makopa and Tambisleaf extracts by recording the scavenging activity in different concentrations. Essential oil.
A substance extracted from Makopaleaves that was evaluated through phytochemical analysis. Free radicals. These are naturally occurring molecules and typically highly reactive that was used to test the antioxidant property of Makopa and Tambisleaf extracts through DPPH assay. Phytochemicals. In this study, phytochemicals are the different medicinally active substances found in the extract of Makopa and Tambis leaves based on the analysis made by Godofredo Stuart last 2017.
Methodology
The researchers examined the antioxidant property of two Syzygium species on different concentrations specifically 20%, 50% and 100%. The samples of Makopa leaves and Tambisleaves were collected and extracted through ethanol extraction at the Adamson University Technology Research and Development Center (AUTRDC). Extract collected from both samples were subjected to DPPH (2, 2-diphenyl- 1 -pircyl-hydrazyl-hydrate) Radical Scavenging Assay analysis at the Institute of Pharmaceutical Sciences at University of the Philippines- Manila. In accordance to the analysis performed, this research resulted in statiscal data. The data collected from the DPPH Assay analysis was analyzed using the SPSS program and ANOVA was the statiscal test used. The materials needed were 500 grams of Makopaand Tambis leaves. These leaves were extracted using the ethanol extraction method. The leaves were suspended beneath an inverted condenser that has multiple drip points across for even extraction. Also, ethanol extraction method has been proven in University experiments to be the most thorough extraction method available. The extracted sample of 10 ml Makopa leaves and 10 ml Tambis leaves were subjected to DPPH Assay analysis. DPPH assay is the most commonly used antioxidant assay for plant extract. Also, scavenging activity of both samples were analyzed through the detected antioxidants that scavenged free radicals. The results from the DPPH Assay analysis were statistically analyzed using ANOVA. With this, the researchers were able to establish the Antioxidant property of the two Syzygium species.
The procedure used in this research was based on the research study entitled, “Antibacterial and Antioxidant Activities of Three Citrus Leaves Extracts” R. Dulay and M. De Castro from Department of Biological Sciences, Central Luzon State University, and Department of Biology, College of Science, De La Salle University, Taft Avenue, Manila, Philippines published last August 13, 2016. 2. 1.
This research made use of two Syzygium species namely Syzygiummalaccense (Makopa) and Syzygiumaqueum (Tambis). The two Syzygium species were taken from Las Piñas City and Quezon City. The extraction was done in Adamson University Technology Research and Development Center (AUTRDC). After the extraction, the DPPH (2, 2-diphenyl- 1 -pircyl-hydrazyl-hydrate) Radical Scavenging Assay analysis was done at the Institute of Pharmaceutical Sciences at University of the Philippines- Manila to determine the anti-oxidant property of the species.
Data Gathering
The Tambis and Makopa leaves were collected from Las Pinas City and Quezon City. Both of the samples were air-dried for a week and were then brought to the Adamson University Technology Research and Development Center (AUTRDC) for extraction. 500 grams of each species were extracted using ethanol extraction. The extraction method used was distillation in which the leaves were placed in a vessel of boiling water or suspending the leaves above the boiling water so that the steam passes through the leaves. The steam strips the light oils from the plant material and carries it to a chilled condenser where the oil-entrained steam liquefies. The condensed fluid obtained from the previous process will separate into two layers. The lower part consists of the hydrosol, which is made up of water soluble oil and water. The liquid forms a thin layer on top of the hydrosol will be the essential oil. The antioxidant property of the Tambis and Makopa leaves were measured through the varying concentration of each extract. In obtaining the antioxidant property in varying amounts of the Tambis and Makopa leaf extracts, setups of 20%, 50%, and 100% concentrations of each extract underwent DPPH Assay. The stable 2, 2’-diphenyl-1-1picrylhydrazyl (DPPH) radical was used to estimate the free radical scavenging activity of the samples, following the standard method of Dulay and De Castro. A 100 µl of test sample in ethanol was added with 5 µl DPPH solution (5 mg DPPH powder in 2 ml of ethanol) in 96-well microtitter plates. The mixture was shaken vigorously and left to stand for 30 min in the dark, and the absorbance was then measured at 517 nm. The inhibition of DPPH free radicals was calculated. Outcomes of the different setups were observed by the given time in the research design, and the DPPH Assay test was done by Institute of Pharmaceutical Science at the University of the Philippines- Manila.
Statistical Analysis
The collected data were subjected to statistical treatment further understand the results of the experiment. It was also done to show if there was a significant difference between the antioxidant properties of the Tambis and Makopa leaf extract in varying concentrations. The ANOVA Test was used as a statistical tool, through the SPSS Version 20 for Windows Software, for determining the significant differences of the antioxidant properties of the Tambis and Makopa leaf extract in varying concentrations.
Results and discussions
The results showed amounts of absorption and scavenging activity of the two Syzygium species on three varying concentrations, specifically, 20%, 50%, and 100%.
The absorbance property of both the Makopa and Tambis leaf extracts obtained from different concentrationsFigure 7. Graphical Representation of the Absorbance property of the three setupsTable 2 shows the absorption of Makopa and Tambisleaf extract under three concentrations, namely 20%, 50%, and 100%. The said concentrations were based on a similar study entitled Antibacterial and Antioxidant Activities of Three Citrus Leaves Extracts. The positive control used for the DPPH Radical Scavenging Assay analysis was ascorbic acid which is known as an antioxidant. The ascorbic acid had an absorbance property of 0. 273 for 20% concentration, 0. 037 for 50% concentration and 0. 047 for 100% concentration. In relation to the antioxidant property, absorbance is inversely proportional with the antioxidant property which is why the ascorbic acid is an efficient antioxidant since it has low absorbance. The Makopaleaf extract had an absorbance property of 1. 404 for 20% concentration, 1. 354 for 50% concentration and 1. 375 for 100% concentration while the Tambis leaf extract had an absorbance property of 1. 396 for 20% concentration, 1. 428 for 50% concentration and 1. 639 for 100% concentration. The absorbance properties of the two species were higher than the absorbance of ascorbic acid.
In relation to the antioxidant property, the scavenging activity directly associates with the antioxidant property; the higher the value obtained under the scavenging activity, the higher the species’ antioxidant property. The values obtained under the scavenging activity of the two species were far from the values obtained under the positive control, which is the Ascorbic Acid. The Ascorbic acid had a scavenging activity of 87. 692 for 20% concentration, 98. 332 for 50% concentration, and 97. 881 for 100%. The Makopa leaf extract had a scavenging activity of 36. 7 for 20% concentration, 38. 954 for 50% concentration, and 38. 022 for 100% concentration. Lastly, the Tambis leaf extract had a scavenging activity of 37. 075 for 20% concentration, 35. 618 for 50% concentration, and 38. 293 for 100% concentration. It can be concluded that compared to the Ascorbic acid, the scavenging activity of both the Makopa and Tambis leaf extracts are lower.
The researchers determined that there is no significant difference in the antioxidant property of both Syzygium species. The Absorbance and Scavenging activity determine the Antioxidant property of both leaf extracts. Based on the data showed on table 3 and 4, the mean difference of the Absorbance and Scavenging activity of the two Syzygium species treated with three different concentrations is higher than the alpha value of 0. 05 with the significance level of 95%. Therefore, the leaf extracts of Makopa andTambishave no significant difference in its antioxidant property in varying concentrations, specifically, 20%, 50%, and 100%.
Conclusion
Based on the data which had undergone statistical treatment, it can be concluded that the null hypothesis is accepted. This states that there is no significant difference between the antioxidant property of the two Syzygium species in three varying concentrations, specifically, 20%, 50%, and 100%. From the results presented, the study suggested that the extracts from Makopa and Tambis leaves showed a low potential as an independent antioxidant. Therefore, its function as an antioxidant alone is not enough to protect the human body from free radical damage. It needs to be combined with other compounds with high rate of antioxidant property to effectively function as a natural antioxidant.
After conducting the study, the researchers recommend to use another type of extraction method to determine if it can affect the antioxidant property of the extracts. Also, the researchers recommend to broadenthe scope of the research by adding comparisons of other antioxidant-rich species endemic in the Philippines.

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