2016-08-15T10:11:42+05:302016-08-15T10:11:38+05:302016-08-15T10:11:42+05:30Acrobat PDFMaker 11 for Worduuid:dc35caeb-7c65-4dfe-8424-a9f8b29647ecuuid:871cc6ef-e217-4c94-836d-a42c4be008d22xmlAntioxidant Activity of MAE Extracted Teak (Tectona Grandis LDavid ZhengAdobe PDF Library 11.0D:20160808104331
International Journal of ChemTech Research CODEN (USA): IJCRGG, ISSN: 0974-4290, ISSN(Online):2455-9555 Vol.9, No.07 pp 154-160, 2016
Antioxidant Activity of MAE Extracted Teak (Tectona Grandis L.F.) Leaves Collected from Different Plantation Site at Java Island, Indonesia Joni Kusnadi1*, Estri Laras Arumingtyas2, Dian Widya Ningtyas1, Elina Cynthia Setiawan 1Department of Food Science and Technology, Faculty of Agricultural Product Technology, Brawijaya University, Jl. Veteran Malang 65145, Indonesia, 1Department of Biology, Faculty of Science, Brawijaya University, Jl. Veteran, Malang 65145, Indonesia 2Abstract : Antioxidant properties of teak leaves collected from Cepu, Central Java, Indonesia which was extracted using Microwave Assisted Extraction (MAE) and soxhlet extraction was identified. The antioxidant activity was measured based on total phenol content, DPPH radical scavenging activity and H22O scavenging activity. MAE significantly produced higher antioxidant properties in shorter time compared to soxhlet extraction, however, the antioxidant properties of teak leaves collected from Cepu, was not significantly different from that collected from Blitar and Madiun, East Java. Different types of soil seem not affect the antioxidant properties of teak. Keywords: Antioxidant, Cepu, MAE, Plantation site, Teak (Tectona grandis L.f.). Introduction Antioxidant has been reported to neutralize the effects of free radicals or reactive oxygen species (ROS) that naturally produce as the body's stress response. Many modern diseases including cancer have been identified to be induced by free radicals. The free radicals can damage cells and its DNA, so if it can be scavenged by antioxidant agent the cancer cell could not develop1. Excessive numbers of antioxidant agents have been uncovered to fight against free radicals. Various mechanisms of antioxidant activity have also been investigated. Researchers suggested natural antioxidant as safe and effective free radicals scavenger1-5. Numerous spices and herbs have been extracted in regards to obtain its antioxidant compounds, however research on antioxidant from perrenial plant origin was limited. Teak (Tectona grandis L.f.) has been known as Indonesian most important commercial tree species that has been reported to contain antioxidants in the plant organs6-8. Antioxidant that has been identified in teak leaves were: phenolic compounds, such as quercetin, gallic acid, tannins9. Teak is planted extensively in Java and cover over one million ha and about 8,000-10,000 ha has produced teak wood annuallys of teak plantation in Indonesia : Unit I (Central Java), Unit II (East Java and Madura), and Unit III (West Java and Banten). These units are divided into forest districts. Now, Perum Perhutani has a total of 57 forests. Cepu, which is part of the district Blora (Unit I) produced the largest amount 10. The leaf of teak usually was not utilized and become waste product of teak timber production. There are three unitand the best quality teak wood among other teak plantation in Indonesiaand the best quality teak wood among other teak plantation in Indonesiaand the best quality teak wood among other teak plantation in IndonesiaThe antioxidant content and activity of plants has been reported to be influenced by the soil where the plant grows. Plantation media and fertilizer applied to plant influence the antioxidant concentration12. Total phenolics in arionberries grown with organic agricultural methods significantly higher compared with conventional method. Strawberries grown in organic media showed higher total phenolic, ellagic acid, and flavonols, than the conventionally grown strawberries13,14. Compost has strong impact on fruit quality and antioxidant compounds of pepper plants under field conditions15. The forest district of Indonesia teak plantation has various type and fertility of soil. Blitar and Madiun forest district as part of Unit II, geologically located in the Old Volcanic Metallogenic, and sediment rock from Arjosari formation16. This areas were quite fertile because of the high mineral content such as kaolin, limestone, bentonite, manganese, feldspar, iron sand, copper, ball clay, onyx, calcites, zeolites, volcanic rocks, sand, trass, gold, piropilit, chalcedony, and other ore minerals17 and the addition of volcanic ash from Kelud Volcano made the areas even more fertile. The annual rainfall of these area is 1892 mm and 3 months dry season18, on the other hand, Cepu forest district as part of Unit I has the annual rainfall of 2062 mm and 4 months dry season. Cepu Teak forests located at the altitude of 30-250 m above sea level, sloping contour conditions, partly rocky (limestone), slightly hilly. The soil type is Latosol, Grumusol, Mediterranean, and Alluvial derived from igneous rocks, cement rocks, sedimentary rocks, margel, volcanic tuff, hard limestone and volcanic bases tuff19. In the previous times teak leaf antioxidant compounds have been extracted using conventional methods, such as maceration and ashwaganda6 and sohxletation7. Microwave-Assisted Extraction (MAE) can help improve the efficiency and effectiveness of extraction with a thorough mechanism of cell destruction by microwaves20,21. MAE has been widely used to extract a variety of ingredients, such as ginseng22, Galium humifusum and G. tinctorum23, Cypripedium arietinum24, tobacco25, Magnolia officinalis26, Angelica sinensis27, Sylibum marianum28, Pueraria lobata2930. The usage of MAE in this study was expected to improve the effectiveness and efficiency of antioxidant compound extraction of teak leaves. Comparison of antioxidant activity of teak leaves collected from Blitar, Madiun and Cepu which has different soil type, annual rainfall and dry season period was done. Material and Methods Fresh Teak leaves were harvested from Perhutani KPH Cepu, East Java, Indonesia. Fifty grams of shredded leaves (1 cm width) were extracted using various method of extraction by ethanol 50%. All chemical reagent used in this study were analytical standard (E-Merck). MAE was conducted by modified home microwave oven Samsung E45 for 2 minutes (power 80 watt). Extract was evaporated in vacuum condition (40°C) and preserved in 2°C. Extraction was conducted 9 times for each method, the extracts were observed for its antioxidant properties in triplicates. The total phenolic content of the extracts was determined using the Folin-Ciocalteau method31. The DPPH scavenging activity of Teak leaves extract was examined using the method of32. The electro-donating activity examination was done using modified FRAP assay32. Results Comparing the antioxidant properti of teak leaves extracted with MAE and soxlet extraction, there were evident that MAE could increase the concentration of extractable phenolic compound from Teak leaves (P<0.0001) (Table 1). The phenolic compound content of Teak leaves extract showed linear relationship with other examined antioxidant properties. Alkaline conditioning of Teak leaves phenolic compounds by Na2CO3 brought the changes of dissociated phenolic proton into anion. Hence, those anionic antioxidant compounds reduced Mo (V) ion of polyphosphotunstates-molybdates inside Folin-Ciocalteau reagent to produce blue colour. Teak leaves antioxidant extracted using MAE showed better activity of DPPH radical scavenging than that extracted using soxhlet (Table 1.). Phenolic compounds of Teak leaves changed the colour of these stable radicals into colourless or pale yellow. This color change was brought by DPPH radical conversion becoming diamagnetic molecules by receiving electron or hydrogen radicals. Antioxidant produced by soxhlet extraction indicated lower activity of H2O2 radical scavenging compared to MAE (P<0.0001). Table 1: Effect of Soxhlet and Microwave Assisted Extraction Methods to the Antioxidant Characteristic of Teak Leaves
Table 2: The IR Spectra Maximum Absorption of Quercetin from Teak Leaves
MAE
Frequency (cm-1)
Functional Group
Transmittance (%)
3448,72
O-H
9,88
1666,49
C=O
22,55
1635,64
C=C
20,36
1388,75
C-OH
25,62
1249,87
C-O-C
26,08
Quercetin has been known as the antioxidant component which is effective in preventing cancer. In this study, quercetin was identified by FTIR according to Nayeem & Karvekar method33. The spectra showed that Microwave-Assisted Extraction method also could extract quercetin from Teak leaves (Figure 1). However, the transmittance value from Teak leaves extract of MAE lower than of soxhlet extraction (Table 2). [
Transmittance
(%)
Wave Numbers (cm-1)
Figure 1: Spectra of quercetin using FTIR Figure 2: Total phenolic Content of antioxidant extracted from teak leaf grows in Madiun, Blitar and Cepu. Comparison was made among each extraction methods. Figure 3: H22 of antioxidant extracted from teak leaf grows in Madiun, Blitar and Cepu. Comparison was made among each extraction methods. O scavenging activityFigure 4: DPPH radical scavenging activity of antioxidant extracted from teak leaf grows in Madiun, Blitar and Cepu. Comparison was made among each extraction methods. Comparison between the antioxidant activities of teak leaves collected from Central Java (Cepu) and East Java (Blitar and Madiun) based on the total phenolic content (Figure 2) and H2O2 scavenging activity (Figure 3), showed no significant differences especially for the antioxidant that was extracted using MAE. However the antioxidant property of teak leaves collected from Cepu measured using DPPH scavenging activity (Figure 4) was significantly lower than the other site of teak plantation. Discussion Teak leaves extracted by MAE gave higher activity than some previous studyusing spectrophotometry method. It was revealed that MAE could give increment against activity of Teak leaves electron donation (Table 1) (P<0.0001). 6,7,31. In this study, the electron-donating activity assay of Teak leaves extract was implemented by Ferric-Reducing Antioxidant Power method32. By the same manner of electron transfer, Teak leaves antioxidant also showed reducing activity towards Fe3+ from FeCl3. Blue complex gained from TPTZ and Fe (II) binding was measured Hydrogen peroxide was known as one of cancer main cause that leads to lipid peroxidation and DNA damage. Therefore, Teak leaves extract also expected to suppress radical formation (OH*) of hydrogen peroxide in the presence of oxygen and water. Previous studies showed that quercetin is the one of main antioxidant compound from Teak leaves obtained from conventional method of extraction. Researchers revealed that quercetin has good ability of anti-cancer activity. Special characteristic of quercetin absorption frequencies happen on 3409–3144 cm−1 (O–H); 1667 cm−1 (C=O); 1610 cm−1 (C=C); 1381 cm−1 (C–OH); and 1264 cm−1 (C–O–C) 33-35. Irradiative heating mechanism of microwave, especially towards moisture of plant material, expected to give overall rupture throughout the cell materials. Frictional heat induced by ionic conduction and dipoles rotation leads to internal moisture vaporization21. Excessive internal pressure of cells caused by water vapours pushed the cells membrane out to swell and rupture. The temperature increment also damaged the structure of polyphenol oxidase, thence this undesired enzyme lost its functionality. Conversely, polyphenol oxidase could not be inactivated by soxhlet extraction due to its lower temperature condition than MAE. By relying on appropriate polarity between the solvent and the solute, the mass transfer process of target compound also limited by its equilibrium concentration36. Therefore, prolonged extraction duration was needed in order to obtain whole extraction. In this case, soxhlet extraction was considered as less effective and less efficient than MAE. Microwave-Assisted Extraction could extract the antioxidant of Teak leaves, which possessed higher total phenolic content and other antioxidant activities compared to soxhlet extraction. Quercetin also found in Teak leaves extract produced by MAE. From this study, it was indicated that novel extraction method gave better performance of Teak leaves extraction while shortened extraction time and facilitated polyphenol oxidase inactivation during the process. This study may attribute to natural medicine development of cancer. The effect of soil type and fertility on antioxidant content and activity of plants did not confirmed in this study. Cepu has been known as the biggest producer and the center of the best quality teak wood in Indonesia11. However, the best quality of teakwood was not revealed the quality of antioxidant content in the plant. Two of antioxidant property used in this study, total phenolic content and H2O2 scavenging activity did not show the differences between teak leaves collected from Cepu with two other sites; however DPPH radical scavenging activity of antioxidant extracted from teak leaf grows in Cepu significantly lower than Madiun, Blitar and Cepu. Most secondary metabolites, including phenolic compound as the main antioxidant compounds in teak leaves, was increasingly produced when plant subjected to stress condition. Generally Cepu has lower fertility of soil, slightly higher annual rainfall but longer dry season compared to Madiun and Blitar 17-19. This condition may caused more stress to teak plantation in Cepu compared to those grown in Blitar and Madiun, however the differences of soil and climate showed by those areas was not significant enough to cause the differences in the antioxidant produced. Higher rainfall experienced by teak plantation at Cepu may suppress the potential of low fertility and longer dry season to trigger higher production of antioxidant.Conclusion MAE significantly produced higher antioxidant properties in shorter time compared to soxhlet extraction. However the antioxidant properties of teak leaves collected from Cepu, was not significantly different from that from Blitar and Madiun. Acknowledgement This research was supported by PHBI Batch II LPPM University of Brawijaya . The authors thank to the Perhutani KPH Cepu, East Java, Indonesia for providing teak leaf material. References
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